THE AECfifiMESM 0F PEREODATE OXIDATION OF SlMPLE SUGARS Thesis fat flu Dogma of 9h. D. MECHIGAB~5 STA?E UNNERSITY JQE-m Christensen 1956 Tl‘l an M) p. L I B R A R Y Michigan State 1 University £2. £4 SYEYE U331". RS? :Y W M gas: LANSING, MIG-NW THE MECIQNIS! OF PERIODA'I’E OXIDnTION OF SIMPLE SIGARS By J9hn.Chriltoncon A THESIS Submitted to the School for Adv:ncod Gradustc Studicl of‘flichigan Stat. Univuraity of Agriculturc ind Applied Science in ptrtill fulfillment of the requircnontl for thc dogruo of DOCTOR a! PHILOSOPHI Depu'bont of Choliltry 1956 i" / (I 1/: Z Hwy/Q wummmm Tho outhor Itch" to uproo- his uncor- opprooiction to Dr. John C. Spock, Jr., for hi: inspirttion, interact, gnidtnoo , tad counsel throughout the come of this involu- gation. He also Idaho: to thank tho Michigan State Uninufly Fund for the gen-rou- grant of In Alumni Pro- doctoral Fallon-hip which undo 1t pouiblo to cmy on this inundation. THE 14me G? PRIODATE OXIDATIOII CF 3mm: WGARS By John Chrieteneen Al ABSTRACT Sub-1““! to the School for Manned mutate Shaun of Elohim State University of Agriculture and Applied Science inipertiel fulfillment of the require-onto for the degree of mm W PmOSOPH Deperfieent e! Che-late: rear 1956 ABSTRACT The oxidetion of Q-glncoee w periodete hee been inventigated with reopeot to nechenien. A epectrophotoeetric method of following the change of concentration of periodete by meaning the opticel density et 222.5 nillinicrone he: been developed. The nechenien of the periodete etteck on g-glecoee ha been found to coneiet or e well "at of undue etteck on «11911:: gluooee eccaepenied by oxidetion of the cyclic fore to give 240ml glycereldehyde eater. hie eeter then llovly hydrolyeee en the rote-determining step in the reection, £013.0de by further uidetien of the glycereldehyde. A lililer re- ection hee been found to teke place with D—Arebinoee. the reeetien of 1,3odiketonee with periodete hee hem inveeti- ated end four 1,3-diketonee have been found to meet (acetyl ecetone, 1.1-dineflvlcyclohexen-l, 3-dione, Hethyl-Lh-pentedione end 1,1;- diphewl-ld-lmteflona). meee involve both cyclic end ecyclic eanpoende. me reection hee been found to produce no tonic ecid or fomeldehyde , but it does produce carbon dioxide from nnenhetitnted 1,3-diketonee. no reaction ie more rapid with periodic ecid then with eodien periodete. l nechenien for the necticn hee been propoeed. the etoichianetric retie of periodete to nnenbetitnted 1,3—diketonee heeheenfoendtobe ebont ton-tome. no reaction of periodete nith grg-glycereldelvde he: been inveetigeted. In reeotione involving exceee glycereldehyde end in iv which final production of tonic ecid and formaldehyde ere determined analytically, e method of calculating glycolaldehyde end glyoxel production ee well ee glycereldehyde consumption has been devised. he method has been applied to the tank of finding the relative rates of etteck on the carbozvl-cerbinol bond (01-02) end the glycol bond (C2-C3) in glycereldehyde. me rete of the tiret bee been found to exceed that of the letter he et leeet e i‘ectm' of an. A damage in coneentretion of e.’ glycereldehyde-periodic ecid reaction mixture produeee en increeee in rete of the nectien, pertienler]: ct the initiel etege. A decreeee in pH bole! thet produced by the periodic prodnoee e decreeee in rate during the tint etegc. Addition of equivelmt neonate of eodim hydroxide poducee e decrease in the final etege or the reection. he depelynerieetion of the diner of §,}_-g].ycereldehyde bee been invcetigeted at 0° 0 end the equilibrium constant hee been found to be 1.0. um or CONTENTS Page mommml‘.........0......OOOQOOOOOOIOIOOCOOOOCCCOOOOOC....9‘ W METHODS AND DAnOOOOOOOOOQOOOOOOOOOOO0.0.00.0...0000 Apparatus and Materiela..u...uuu"a...".“nun-u...” Imam“-eeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee rig-tends.O...3...‘OCCOQCCCCOCCOCOCOCQOOC0.0...’........... Ultra-Violet Absorption Method of Determining Periodic 101d... Interferencee................o...............c.....ccoc.... Determination O! ebeorption MIeeeeeeeeeeeeeeeeeeeeeeeee Sundry 0f poeeible 9m-eeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee Application 0 the ebeorption method of dcterninetien to 1.75 X 1 M sodium periodete ”lauMeeeeeeeeeeeeeeee 32 Effect of eodinn bicerbonete on Wine or periodic ecid 33 Experimental methOdeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 35 Oxidation 0f D‘GlfiOOSBeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 35 Reaction of 0.025 n periodic ecid with 0.0th n D-glncoeeu 35 Reaction c: 0.0875 n eodim periodete end 0.012511 D—gl‘ll-COIO.eeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee bl Oxidation 0f D’Winmeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee ’47 Reectione of Feriodic Acid with Verioue (hmpoende which are 'Ot anguleeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee SO “IIOOIBldehydOeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee SO GlycerOleeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee SO Methylene glyOOIeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 52 icetonyl “GMeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee Sh KW]. acetonetatfleeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee Sh Loetyl acetoneoeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 5h Dhnedone (1,1-dimethyl mm3,5‘dione)eeeeeeeeeeeeeee 6O Methyl MSW]. acetone-eeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 6’4 310”]. ecetyl ecctophenoneuu'u..uu.................u.. 6,4 EWOMWdOeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 66 8&013’191338! Q’— .4 DWMOOOOOOOOOOCQOOOOOOOOOOOOOOOOOCOOOOOOOOOOOOOOOOOOOOOOOOO " mmeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee n fi‘mbimneeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 82 ,3‘D1k0t0neleeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 83 lbWOO’ddfih’dfleeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 86 General Reaction."nun”...uuc.nun"”cu-nu..." 86 Development or reletimuhipe when mereldehude ie pneent in excelleeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 88 Eff.“ O: I Chillfi in weeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 91 vi 0'... '0’. TABLE OF 0mm - Continued Effect of e change in cencentretion without change of the glycereldelvde t0 periodate utioeeeeeeeeeeeeeeeeeeeeeeee Effect of increasing glycol-aldehyde concentration. e e e e e . e e e e Effect 0! increasing periodete concentratiw..uu. eeeeeeeee WOOOOOOOOOOOO0.00‘OQ.OOIOOOCIOOOOOOIOOOOOOOOOOOOOQQOOOOCOOOC LIST OF mmm.OOOO'OCOOOIOU00.00.000.600.00000000DOOOOOOOOOOCO vii Page 92 93 9h 95 97 INTRODUCTION the Heleprede (1-3) reection, which involves cleaving e chain between two carbon etue, eech of Which carriee either e hydroxyl, e carbonyl or en nine poop, hee been need extensively for determining the etructure of unknown couponnde end for quentitative analytical determinetione. mece neee are based on the eecmption that e periodate reecte quentitetively with the 1,2 mp. listed end um e reaction doee not teke piece eith other grouping. the reaction oen only main dependable ee en enelytioel tool if the conditione under which etoichio- netric reectiene teke place ere known end can be utilised. m1. re- quiroe e knolledge of the nechenicme of the reectione. In the nee of periodetee for either of the purpoeee nentioned, the progreee of the reaction ie usually followed either by deteneining the decreeee in periodete or the increase in come product, or products, etch ee for-aldehyde, formic ecid or occuionelly come leee cannon product. It ie therefore eeeentiel that the reection proceed etoichio- metricelly. A «arch of the literature has disclosed e number of anomalies with reepect to the reaction uhich cen imelidete new of the reenlte obtained w the nee of periodete end eleo e mmber of dieegreemente heteeen inveetigetcre on lettere of £3111de Meme. It '0 therefore decided to inveetigete further the mechanism of the reectien between periodic ecid end the einple eugere. Haw eontradictuy statements with regard to the influence of various factors on the reaction has been noted. With respect to the silple glycole, Rico and troll (1;) found that with ethylene glycol and pinecol the reaction was acre raid in acid than in slimline solen- tion and that the reaction with pinacol was a general acid-base catalysed reaction, although the variation of rate constant with pH produced a complex cum. Price and Knoll (5) found the rate constants for ethylene glycol essentially independent of pH below 7 and. falling off rapidly above pH 1. Beidt, Gladding and Purvee (6) reported the reaction with ethylene glycol as acre rapid at lower pH's, falling off aptopHS, thenrisingto approximatelypflfl and assumingverylow values in am alkaline media. Duke and Elwin (7) predicted the rate for ethylene glycol should be independent of fl in the range pl 3-7 because increased acid catalysis was offset by decreased ionisation ef the periodic acid. Exist and Benton (8) foumd the maximum rate eoefficients for the ethylene glycol-periodic acid reaction at pa 1: with a decrease on both sides and a very sharp decrease above pa 7. Taylor, Soldano and Hall (9) for most glyccls describe a broad pH range Demon 2.5 and 6.0 reducing maxim reaction rates and a decrease at either end. For glucose, luau and Mall (10) found that pericdate was coco sued faster in buffered acid and in neutral solution than at pfi 9.7. week and Porist (ll) found the reaction with glucose slower in acid than in bicarbonate buffer. Fleury, Courtois and Bieder (12) ondised aldohexoees acre readily at pH 6.5 using a phoefiate buffer than at p8 1.6, but the polyhydroaql canponnds, mannitol and sorbitol were caddiscd more rapidly at pH 1.6 than at 6.5. Greville and florthcote (13) with di, tri— and tetra-g-methylated glacoses detected a decrease in rate with decrease in pH, but a few canponnds which did not have the expected structure for normal reaction showed abnormal reactions and oxidized active hydrogens faster at pH 5 than at pH 7.5. Neumuller and Vasseur (1h) with maltose, melibiose, nefivl-d-Q-glncoside, yeast, glucan anri dextran found a second or mrooddstion stage having a minimum rate at p?! ‘3 and h. Jeanlez (15) uing metIu'lated sugars found rates faster in weakly alkaline solution. Dell (16) with methylated sugars obtained best results at pH 7.5. Others who found more rapid rates in the alkaline region were Reeves (1‘!) with nonossccharides in sodium bicarbonate buffer; Grangaard, (Reading and Forms (18) with starch and cellulose; Jamie: and Porchielli (19) with starch and hyalnronic acid; Lindstedt (20) with yeast nennsn; Bprinson and Charger)? (21) with tartaric acid) Huebner, Amos and Buhl (22) with various organic compounds containing active hydrogen; Van Slyke, Killer and HacFadyen (23) with hydroxy lysine; McCasland and Salt!) (2h) with minocyolanols; Jeanlos and Forchielli with chitin (25) and glnccssmine (26). with respect to the effect of other ions on the periodate reaction, Taylor, Soldanc and Hell (9} discovered that the rate of reaction of periodates in the presence of neutral salts is increased proportionally to the concentrations of the salts. Bnist and Benton (8) round rate constants dependent on ionic strength and acidity and Huebner, bees and Debi (22) found considerable variation at the sue pH with dif- rerent batters. a number of investigators have reported greatly increased rates and even sane abnormal effects from the use of phos- phate batters. Fleur}, Gonrtois and Bieder (12) with aldoses and Bell and Greville (27) with 3,h~di-9_-methyl-2-glaoose report increased rates with phosphate. ihe latter also report formation of reanaldelvde very late in the reaction and an early production of formic acid. Bell (16) with 2,3- and 2.6-di-Qomeflvl glucose observed over- oxidation and production or carbon dioxide. Bell, Palmer and Johns (28) noted that when the periodate to phosphate ratio was increased J-Q-neum and 2.3,h-tr1-g-nethy1 glncoses produced nor. tomaldelwde and less formic acid and vice'versa. They also noted an ova-om- tion of periodate. 'lhe total yield of formaldehyde and tonic acid was approximate]: one mole per nole of the trinethyl choose, so that when one increased the other apparently decreased. Lindstedt (20) using yeast naman, catalysed by phosphate, found increased rates and overccddation even in acid solution and showed that two thirds of the batman molecule was oxidised to carbon dioxide at pH 1' and 50° 0 then catalysed by phosphate. ’ With a variety of carbohydrates and derivatives he obtained overoacidation and in sale cases carbon dioxide. Mills and Nuthcote (13). on the other hand, found no acceleratiu due to phosphate catalysis on methylated glucoses. In fact, they fund the rate even faster when the solution was adjusted to the desired pa w mm rather than 11v phosphates. inmborotmkmhmroportedpnohugudurinztho course «the reaction. hilt wanton(8):mdnmdrop1npnm In oxoou of glycol m nixed with periodio acid ad which they umod to be duo to the motor acidity of the mum ample: bots-on tho two oaponndl. i firth” Ila! dear-em followed, which was ovi- donfly due to tho doomed.“ of the cuplox and the fond-ion of tho relatively ltrong iodio Acid. At. pH 5 they round n m door-cue in y! followed ‘w o Ilou increase. Price and Knoll (5). tho using othylono glycol, over the angst pH 1-7.5 and 1.04.1.5 found no changes of 153 during tho reaction, ht they did find a chap in the rungs 1.5.10. rickshaw“, Smith (21;) «in: W fund a rim or 1¢3p3unita Wilton-cacti”, butthismoppuronth duotothe mom: liborlted. We and Moot. (13) observed tint in am "Whtod m the rate of oxidation increases with the decree of nothflxtion sad that thorn“ in not dependent on the amber of actual or Monti-.1 bdrm). mo, but rather on the amber oi‘ nothwl radical... For instance, tho mum of periodate by 2 ,3, h,6~totravg~nothl~§« glucose ranch“ Hus nolu, mm, manually, 1h moles or: re- quiredtooxidiuiioupiehlytofonioooidmdfihmuou‘bon dioxide. * ' mawmwode-tmunmouo oi" mummdetominotporiodueinthooolntion. Raini- tho method of Flam? and Longs (29). Eadie! and Novel]. (10) attributed tho immiatenciu to the slow reduction of the intemodiato mile: between periodato and the glycol (gluon in thio caee). I! this ia true, the areenite aethod rould not mum the periodato which had already formed the intermediate complex, ht which had not yet oxidized the glycol. mow aloe found that the eoletiona after reaction Iith excee- areenite liberaud conoiderable iodine epon otanding. They med this to be the result of a aloe reaction of the intermediate couple: with iodide ion: in the olidatly alkaline solution. uch a reoolt would naturally indicate a oorroopondingly larger mention at periodate aa oupared with formic acid produced. Spock and Foriot (ll) aloe oboerred thst poriodato con-mention oo honoured by aroenite reduction appeared to proceed considerably teeter than toenaldehyde tonation. Van Slyke, Killer and KecPadyen (23) obtained a oinilar diocrepancy in roe-king with Whine, but they attributed the results to the fact that their reaction manning in acid oolntion woo promoeing slowly. hen upon adding oodiun bicarbonate in order to produce a favorable pH for reaction Iith oroenite , the reaction with poriodate took place instantly. mg {and the glycol-periodete reaction in oodinn bicarbonate buffer to be alnoot inotantaneono. scheme (30) oxplaino the apparent high periodato coneuption, then titrated w the oroonite method, ao being a remit of oxidation of active hydrogen or of iodinatim by free iodine liberated in the alkaline oolntion. this reaction meat be W ao follow" CH (H (I! (1) . 2-] ' 2-l ' 2—] 50-9-3 - (HO (HO Ho-c-n fl) sol-l we 3-; EH) no-%: 3.3.03 3.3.0 duo 31203 + m l , h—anhydromannitol f In this connection it will be recalled that free iodine in an alkaline solution substitutes for alpha hydrogen in a manner similar to the iodoforn reaction when aldehydes or lcotoneo are present in the reaction mixture. Overoxidation, which may appear merely as a slight overconsumption of periodate without apparently diminishing the analytically determined products orehichminits extreme case involve oxidisingnoot of the products of the normal reaction too arbon dioxide and voter, appears tobeacmonphenanenonitcnecanjudgolythoumberotreferences to it in the chemical literature. Overoxidetions produced by phosphate catalysis has already been comidered. ‘ been, Dmaten, Halsall, Birst and Jones (31) conducted their experiments on end-m determination in polysaccharides specifically to determine conditions for avoiding overoxidetion. Mitchell and Percival (32) working with several methylated fructoses found our- consumption of periodato and less than theoretical yields of formaldehyde in nearly every case. Hiyada (33) on the other hand obtained prac- tically theoretical formaldehyde yields and poriodate canonization with 3,h,6-di-Q~nethyl~2~fmctoso. Doerschuk (32.) found in an oxidation of glycerol-bow ty periods.“ 0.25% of the 01h present. as remit acid. Ie attributed this to enolisetion occurring in the glycol aldehyde Ihich resulted from the first stage of the reaction. his may more likely hm been the result of moi-oxidation of forualdehyde w the periodic acid. Jeanlos and Forohielli (19,26) concluded from their studies on polysaccharides and gluoounino that mroxidatia is a faction of temperaturcaseellasofpnandthatitoanbeolhinatodwmng at 5° 0 or laser. mey also remanded a pl of h-S for maximising overexidation. \ Hetlvlated sugars appear especially subject to maniac as previously noted in atom to them of cad-«ins and lorthcote (13), Bell (16) and sell, med and Johns (28). She effect of lidxt on periodate solutions has been carefully investigated by need and Standing (35). has mm that periodato solutions will reduce to iodeto and finally to free iodine upon sufficient smears to next. Head and W (36) found that when catalysed periodeto reactions with simle organic outpoands such as tee-nit acid, formaldehyde, moms acid, methyl alcohol, oxalic said, ethyl alcohol, acetaldelurde. With a large excess of periodate in height annexe itmrewmhistoeapistslyoxidiu tonic acidincnedsy) othmiseanattorofdayswosroquirodtoprodnoe significant ouidatim. seed (3?). investigating the affect of dsnigm on periodate oaridaticu of alycosides and cellulose, found definite catalyeia w light. However, in the course or two hoar- no appreci- able difference an observed batman reactions run in the dark and in the dqneat. In fact, in six hours the difference was small, but in tom of day: it hecaae very great. He continued his observations over periods as great an 120 dare. The results of ooddationfl extending over this period or time are quite meaningleaa in View or the frequent observation of merofldatim by periodate. Head alao found that a great excess at wicdato tend: to increase mrooddetion, ac Inch no that the curve chm no differentiation between the original Halaprade reaction and cremation. He also found that free iodine is produced when there is insufficient pericdate . Pericdete in also known to oxidize certain calm containing canv- poanda. licclet and Shim: (38) round that periodic acid reacted with methionine and cyetine, month w oxidation of. the calm. Donner and Driekc (39) hm shown that phcxwl-fi—D-thiogluccpyrmoeide is oxidised to e euli‘one, in addition to the noun]. glycol cleavage, and that thin reactiw We: iodine. This iodine caddiecd active hydrogen to ahydrml groom u ahovn in equation {1), thus making further reaction with pericdato poeaihle. i mmber or workers have reported the reaction or periodate with active hydrogen, either with or withont carbon dioxide production. the timiinge of Schwara (30) have already heennenticned. Emilee end Vueeur (114) reported a cecond stage or cndation with maltose, edniler to the exuple in equation (1), in which active hydrogen ne‘oocidieed to a hydroxyl group capable of W oxidation with pod-iodate. 10 Head and Hugues (he) postulate essentially the same results on intemediato canpounds in tho mddation of cellobioso. 12113 is illus- trated as follows: H OH 0 I I I (2) one-p.610 _Q_.,. onc- cno 313335...) R-o-c-mo + moon 9 . n R Huebner 33 5;. (hi) reported a similar oxidation of active hydro- gen in borzvl glucoside and or the hydrogen on carbon 1’ in 20.51”- amydro-Q-qulo-tetmhydroxy butyl) benzmidazole. 3) H- 30 13-on H-G—O—Bo ( 'I I I H-C—OH (no I mo I m o 2 IO): 0 101‘- o . ——-—--—> __....a, 3-0-03 ' mo mo I I I 3-0 11-0 HO-C I I I @1205! 03205 (11203 Bormrl glucoaide Bo- Borrwl radical \ / H-N N \ l C ' 2(1' ,hkanhydro—g-rylo- H~C§ tetrahydroaw butyl) I benzimidazole 30- 0-3 ' o lecherger (M) eleo reported I einiler phenmenon in connection with periodete oxidation of ethyl glucoemte (together with the men-moo of tree iodine) ee chm in equation (1;) coon: ' mom E—C-NEI can; 90°“ v ' . o " (1:) 30-04 31% n—o—mocch; rob 3° o—mocéas H~C-OH mo I 342-03 ’ $720!! 23000:! c ECHO Potter and mm (13) in and group detemutione of starch also report oxidation of the active hydrogen in the tonic acid ester or tutromldahyde. mm, air-t and Jane: (M) found that when on ective hydrogen m oxidised, each as in a methyl made, iodine m tamed. may eleo found thot when mum box» or pcntccyrmccmec tore oxidised thet normal products were obtained. Buckner, Amos and mm (22) epeciflodJJ checked the reaction- hetueen certain canpaunde containing eotive hydrogen with period-to end found that e reaction took place with nelonic acid, tertronic acid, nemalic acid, nonoethyl molonate, nelio cold, diatoxoce, ecctoecetic cud, alphccethylmelonic acid, axelecetic acid, acetone dicerbouorlic acid, citric acid, lectio cam, pyrmc acid, 1,14-anhydroeorbitol and ethyl oxomlonete. new of those produced cox-hon dioxide in the re- ection. Sane were very rapid, u for example anionic acid using 3 .01 equivalents of periodate 3 acetone dioarbonlio acid, h.5 equivalents and ecotoecetio acid, 2.5 equivalents of periodete in an hour. Sigdi‘icantly, they reported no reaction with acer acetone, because it gave of! no carbon dioxide. These reactions were run at 21° C in a solution 0.0h-0.06 H with respect to the substance being oxidized and containing from 1.2 to 1.6 times the molar equivalent of sodium meteperiodate theoretically necessary for complete oxidation. Sprinson and Chargai‘f (21), investigating the periodate oxidation of active hydrogen, proposed the following reaction with malonic acid: (5) moo-ca ~cocn 10E ‘ HOOC-CHOB-COCH IO " oncooon + co . 2 h 2 cue—fl ---—--> A? 002 e HCOOH 'Ihey found that althonfi these reactions as sheer than the nomel Halaprede reaction betssen tee groups consisting of carbowls or hidroxyls, the reactions nevertheless are reasonably rapid and met be taken into consideration in the use of the Helen-ado reaction. rm- instance, tartaric acid consumed he. one to three moles of periodate in loss than one hour, the mount of periodate depending on the con- centration of periodate, p8 and tenperatnre. A most interesting observation was made by Conrtois and Joseph (b5) the reported a slow periodate reaction with dimedone, a 1,3- diketone. 1111s substance consumed 2.67 moles of periodic acid per mole of dinedone in 96 hours. Wain-cu and Bobbitt (1:6) hm recently obtained reactions with a mber of cyclic 1,3—diketones and have proposed a mechanism for the reectims. hey, halever, reported that acyclic 1,3-diketonee did not react. 13 mm and Doisson (In), Nicolet and Shine (38) and Ibouvine and Arragon (148) all attribute the fonnation of carbon dioxide to periodate cleavage of glyonlic acid. ihe first of these groups of sorkers reports that glyoxylic acid is quantitatively transformed into fomic acid and carbon dioxide in 2h hours. However, this possum: takes place at the temperature of a hot water bath, which is definitely shove the tauperature at shich reactions of enalqticel significance my be run. mentors, in addition to the usual Halaprade reaction with periodates, at least two other types of ooddation'secn to exist! (1) Oxidation of so—called active hydrogen. (2) Cleavage of a car- bonyl groupfronacarbowl group, ertoalesserextentfrma. carbinol group. A lmmrledge of these reactions which involve unusual periodate oxidations is of great value in analytical work involving periodeto oxidation of cubohydrates, so that periodate consumed in these reactions will not be attributed to the usual Helaprade re- actions. In addition to these reactions there are others which are more difficult to classify. Nicolet and mm (38) report a slow reaction with a variety of amino acids containing only amino and carbonl poupe. mis perhaps can be classified under type (2) above, inasumch as the amino you}: would be susceptible to lvdrohsis. The reactions of periodate on simple organic compounds such as methyl and cum alcohols, as reported by Head and Hugzes (36), would also be difficult to classify as one of these two types. Another 1!; reaction difficult to classify would be the one reported by Rsmachandra and 3am (h9) of the oxidation of 1,2-dihydrOJU-3-anthraquinone sulfonate, uhich is the same product produced by mild oxidising agents such as alkaline ferricyanide. With respect to the kinetics of the reaction between periodate and the glycols, Price and troll U4) worked out rate constants on the basis of a second order reaction for pinacol. Price and Knoll (5), in addition worked out second order rate constants at various pfl's for ethylene glycol and for cis- and trans-cyclohexene glycols. Duke (50) was able to demonstrate for the first time by working at relatively him glycol concentrations that the kinetics of the periodate-glycol reaction are canpatible with the mechanina in which fox-nation of a periodate-glycol complex is rapid and reversible, whereas transfomation of this complex to iod ate and fomaldehyde is the rate determining step. fast slow Periodate e glycol g‘ periodate-ghrcol —-——-—-¥ iodate + complex formaldehyde Taylor (51) expanded Duke's work and found that the rate constants for the second order reaction increased as the reaction proceeded, especially at low temperatures and equal concentrations. Upon increas- ing the glycol to periodate ratio, the rate constants decreased markedly. Duke and Bulgrin (7) have worked out a series of rate constants for the methylated ethylene glycols in Which frat zero to four methyl ms were substituted for the hydrogens on ethylene glycol. 15 lbs! drew the conclusion that the mechanism involved an active inter- mediate and that in all except the tetranethylated compound (pinaccl) the rate determining step was the disproportionation of the complex. In the case of pinacol the formation of the couple: was the rate determining step. Ibis was seamed to be due to the steric hindrance of the methyl groups. mist and Danton (8) have worked out a series of rate constants as affected by pH and have drawn the conclusion that the attacking species of periodate ion is monovalent. This ion could be either 10,; or H1110; with the fencer as the more lone-l choice. Taylor, soldano and Hall (9) have given evidence from the pH effects that the attacking species must be a malent ion in that the calculated pH-rate curve based on a monovalent ion agrees very well with the experimental curve. However, these calculated rates when applied to pinacol and some carbohydrates do not correlate with experimental data. Criegee, Kraft and Rank (52) early applied the theory of a lead - tetraacetate ester intemediato with glycols to the periodate complex. his idea that the intermediate is an ester of. the general structure H-S-O\ H-c-o’ I 101-1- + H. has been quite generally accepted by workers in the field. Heidt, Gladding and Purves (6) have attemted to give a mechanism for the formation of the intemediate in toms of molecular models. 16 Smith and Duke (53) have amen a probable electronic mechanism for the reactions involving the cleavage of the carbon chain. an. m... to the nethed of attack to the pericdate ion upon the glycol Ming, Price and Knoll (5) have postulated a back no. spproaohortheperiodateionandinshiohthereactiontcningthe intermediate coupler takes place in two stages. I: the last stage is seemed to be the rate controlling step it wosld ease with the data presented by a member or workers, nsnelythat the cis-glvocl grouping reacts faster than the trans-glycol. Price all! Knoll (5) found this tobetrne inthecase ofthe cis- sndtrans-vcloheoccne mole. Henry, We and Binder (Sh) in comparing the rates or the reaction of periodate on sldosee and polralcohols found the following relative reactivities: 5mm) Sorbitol> Hannoee > Oelsetoee > olooooo his armament corresponds with an arrangemet in tens of decreasing numbers or adjacent carbon atom with the one configuration. McCaslsnd and auto (2h) experimenting with slinomlsnols fonnd the ciao advantage in reaction rates anch less pronounced in the case of reaction with periodste than with lead tetraacetate. With ois-2- ninocvclcpentanol the reaction was four tines as fast as with trans- canponnd, but no: cis-2-ninocyclohexanol the distinction n. not pronounced. In sue cases the trsne-cunpoand even reacted taster. Certain ring structures have been used to duonstrate cis- trans- effects. , om», Davis and Hilbert (55) found that ens-mp- zlncohrsnose (I) does not react with periodate and Alexander, Dialer and Hehltretter (56) similarly toand that 0: -l,6-enhydro-Q-galacto~ Menace (II) does not react either. 17 18 The assumption has been that it in because of the truma- positions togethar with tha two locked ring: which effectively prevent any in— mariaation. noun-nan and Smith (57), carrying this idea farther, mthoailed tho oanpounda L-tm-eitan (III) and erythritm (N) which both reacted with poriodata. nevertheless the L-threi'tan reacted the almost, particularly with periodic acid. Both at then reacted simi- with the pariodio acid than with the aodima periodate, and with the acid the difference between tha an more mowed. Jackson and Endaon (58), on the other hand, prepared 1,6-anhydro- B-glucopyranoao (V) which reacted with the periodata. 'lhia comma also ha a doable ring straw and trans- grouping of hydroula and yet undergoes a reaction. On this haaia amino and lm-thooto (13) are or the opinion that it in not the trana- configuration that inter- turn with tho reaction. lunar, thay apparantly do not take into cmideraticn the fact that in (V) the ring We: inch carries the hydroxyl ml in nix-amend, what-cu in (I-tIV) the ”from; ring! are timber-ed. 1119 six-membored ring in more puckered and pamita greater Mutant of tho atma than the more rigid and planar Mabel-ed ringaaamobaomdinthe-omofuccaaland and saith (1h). . mm (59) found that tho Liv-lacuna of anomaeohu-io acid (VI) did not react with periodic acid in one how, when“ tho gluooaacdiaro- 3,6-1aotono (VII) reacted radii: with periodic acid. Again the W1 groom are on a rigid numbered ring, me]: could prevent the imeriaation or trana- mm. l9 \ 0 on on 0-03 I H H-c-OH ” u o 0-3 HO-C-H | i 3- 0-0:! 00011 0 t 11- con ' c-o VI VII Taylor (51) discusses the effects of reactions on compounds (II-IV) and dram the conclusion that the hindrance is caused by a field effect of the oxygen, iherebw the oxygen repels the negative periodate ion in a' backside approach. min is a possible explanation and in the case of a six-membered ring fomed by the bridgehead omgen, such as in (Y), the oxygen being further removed, could have less effect. In most of the previous discussion of mechanim the emphasis has been mainly on the reaction between periodate and the fundamental glycol groups. Hughes and haven (10), hoverer, mm the problem of the mechanism in thereaction or periodic acid on sugars such as glucose. than they measured the progress of the reaction between sodium periodate and glucose by sodium thiosulfate titration of the iodine liberated Ira excess potsssimn iodide in acid solution, they {and a fairly close correlation between periodate consumed and formic 20 acid produced, except for a small discrepancy at the 'beginning of the reaction,‘ as they call it. Therefore they postulate a very mtomatic attack m the glucose molecule, cleaving first the are, bond, then in succession the 02-03, Coy-v6)4 bonds, etc. to the end of the chain, thus liberating fonnaldclvde as the final product. 'mis appmntl: would eaqilain the lag in the production of fonnaldehyde , which they found according to their data. 'mcir first neasurment was taken after 22 hours had elapsed and therefore they really did not have the data at the beginning of the reaction. In View of the possibility of a tendon attack of periodate on glucose it some strange that the attack on the 01-62 bond should be the exclusive node of attack. One of the reasons for mdertaldng this research problee was to mrther dieck this phenomenon and if possible to detcmino the actual mechanism or the principles involved in preferential attacks on sugar molecules. Itisagenernnyacoepted fast thatonlynextrenolyuall percentage of we glucose molecules in a collation are preeeat in the open chain aldehyde fora. ‘ 'roassanethatonlytheopenchainfosareaotewithperiodate would seem rather pres-whens in View of the fact that polysaccharides with locked rings react practically as fast as glucose shamr- sollble. Monolith, a number of investigators have unused that possiblyafosnie esternsybe famed inthe re'aotioninthe following 21 H— H 114: 30003 9 l mom 0 + t 31110 - Hon 21110 6 no- on o o o o 23on ( ) ‘ -—-—lL-) s —__> e ”a n-c-on B—C 3-0 * c A a B ' C 3-0 11.5 H-G—OH HCHO e e ' 3.0.03 3-6-0}! 3-53-03 I I H H H +amm , Hugues, himself, in collaboration with Head (10) assumed a similar mechanism in the mdation or cellohiose and whereby it would produce a tonic acid ester. The hydrolysis of such an ester could be the rate determining step or the reaction. Harrison, Inpyer and Orton (60) seems the formation or such an ester as the reason for obtaining less than the theoretical mount of formic acid from maltose. Nemller and Vssseur (110 and Halssll, His-st and Jones (“4) also named such an ester. Meyer and Rathgeb (61) fond evidence or it in determining teminal end groups in starch. Barker and hith (62) , by starting with glucose containing a methyl or a nethenesnlfowl (neayl) mp substituted on carbon 3 and oxidising it With sodium metsperiodate, obtained h-i‘oxuyl-B-methyl- D-arabinose or hefwl-B-nesyl-D-arebinose, respectively. mess We were quite positiver identified by carbon, hydrogen, methyloxy and {session nurses as well as by physical constants, derivatives and infra-red spectra. Smith (63) later used the periodate reaction 22 with Meryl-D-glucose in a preparative reaction in which the formyl ester us need for the preparation or ribose. Schop! and Wild (61;) in oxidizing glucose with sodium periodate found that three moles of periodate reacted rapidly, but only two moles of formic acid were readily available for titration with sodium 113de- ide. A third mole of sodium hydroxide was slowly consmed over a period of four days. his was explained by step A in equation (6), shich consumed three holes of periodate and produced 2 moles of formic acid plus one sole of the tennis ester. mil ester than Wed ever an extended period, thus really being the rate determining step in the reaction. The: performed this reaction on a preparative scale, isolated the ester, determined its physical properties, hydrolysed it and identified the dimedene derivative as that of glyceraldehydo . men they reacted one sole or sodim periodate sith one mole of glucose they recovered two-thirds of a mole of glucose. his some to indicate thatoncetheringiscleaved, thebondsintheopenchainarenore readily available for reaction than the bonds in the cyclic etmcture. his mechanisn fits the experimental data determined by ladies and Nevell (10) inamch as by the time they made their first analysis of file reaction sixture, a large proportion of the ester would have had an cppa'tunitv to hydrolyse, producing almost a sole of tonic . acid per sole of periodate. 'Ihe remaining unhydrolysed ester evidently produced the mall discrepancy that they noted. It was the purpose or this investigation to elucidate the mechanic: EV loans of uhioh the Modate attack on glucose produced formaldehyde 23 at a slower corresponding rate than periodate was oonsmned. During the course of the investigation the formation of a {crawl eater hem apparent and the idea was soon corroborated by other investi- gators who have been cited. To investigate the further attack on glucose aid other simple sugars the investigation was extended to in- clude relative rates of reaction with various bonds in intermediate mducts of glucose oxidation, particularly in glyceralderwde . ext-mm mom AID mu Apparatus and Materials 5%. A Boolean, aedel DU spectrophctmeter with 1 ea. quarts cells was used for steaming ultra-violet absorption and a loom, model I spectrqmotaeeter with 1 a. Goren cells for measur- ing absorption in the visible range. Materials. me periodic acid, iodic acid and soda iodete used were produced w the 0. Frederick nun (helical do. no tonic ma, tWMe, "dill Mom. sodiu «mu, W1 an! Mm acetate acre 0. P. grade. me Q-glacose used for these experiments melatienalm'eaaet Standardsdextreee. meg-mensw- marmoman-m. Iheaeeteluaselles Ge. product. the ehrcaotrepie acid (MSW-2, 1+nephthalenedisaltcnic acid) was Hathescn Om, practical grade, and m purified I" recrystal- lisation true 501 ethanol-eater airm-e. ihs glycolaldehyde was prepu‘ed by the aethod of Fischer and frauhe (65) {re- dihydron aaleic aciduhichnsintu'oprepmdtrutartarieaoidwtheasthodo! W and Feldnan (66). mmwimmmnmm mmt. ItIasre- distilledandthetrectionbeiliuintheraaseus~136°nscolleeted. Its refractive mm a 18.3. «a 1.1628. mm). acetvl acetone m prepared Ira new}. acetone and methyl iodide by the method or mane, Deckhan and men- (67). Phervl acetyl acetophenone was 25 prepared from etivl phewl acetate and acetophonone IV the maiaen reaction as developed by Below and Grotowelq (68). Glycol-aldelvde, unleaa otherviae etated, was prepared w the I'Ormie Syntheeee' procedure (69). the last glycoraldelwde oaod m a Intritional Biochemicals Corporation product. he glyonl need no a technical grade 30% solution produced by Union carbide Corporation. Ultra-Violet theorption Method of Determining Periodic Acid In order to determine the progrese or periodic acid oxidation it no neceeeary to have on analytical method for following the men- tretion changes of periodic acid which would not alter the conrae o: the reaction itself as the areenite method hae been known to do. To achieve thia, ultra-violet absorption curves of 0.0001 H periodic acid, 0.0001 R iodic acid solutions and We of both which totaled 0.0001 14 concentration of acid me plotted with the Becknan, model DU spectro- photaneter. In all experimental work fresh periodic acid eolotione were we daily. Interferencee. To detemine whether tomcldohyde, formic acid or the carbon dioxide in ordinary diatilled water would interfere by Ileana of their om abemptior, a preliminary amendment uaa performed. No 0.0001 2‘! periodic acid solutions Vere made, one made up with ordinary dietined water and the other with doubly dietilled carbon dioxide-tree water. ' 26 A third 0.0001 1! periodic acid solution containing fcmio acid to the 0.00009 14 concentration and a fourth containing formaldehyde to the extent of 0.000018 1'! concentration were made up. Those concen- trations of fomic acid and formaldehyde are approednately the maximum edpected when any reaction between periodic acid end glucose is com- plete and has been diluted to 0.0001 K concentration, based on the maul periodic acid. nae mdm absorption occurred at 222.5 m, which would then appear to be the logical point for measurement of the optical density of periodic acid solutions. A comparison of optical densities at 222.5 mu gave the following results: 0.0001 1! Periodic acid in ordinary distilled water 0.982 0.0001 1‘! Periodic acid in ordinary distilled water 1.02 0.0001 14 Periodic acid in ordimry distilled water 1.02 0.0001 1! Periodic acid in doubly distilled cog-mo water 0.996 0.0001 H Periodic acid with 0.00009 H formic acid 0.960 0.0001 H Periodic acid with 0.000018 8 formaldehyde 0.981 Apparently none of these canponente, carbon dioxide, ionic acid or fomaldelvde have an appreciable absorption at this concentration. heir effect, if any, ia to lower rather thu to increase the optical density. Of these, the only one which has an effect beyond the experimental error is that of fomic acid. Dixon and Lipkin (70), who dm10ped e smeadxat einilu' spectro— photanetric nethod for periodate, report that in the 1mm been of the spectrophotaeeter fomic acid is oxidised by netaperiodete to 27 carbon dioxide. This could account for the slight decrease in optical density of periodic acid solution containing formic acid. Dixon and Lipkin, however, neglected the absorption of the iodate, Ihich was pro- duced in the reaction. This neglect introduces an error of about 12-13% in the analysis. Determination of abeorgtion curves. From a 0.0001 H periodic acid solution and a 0.0001 11 iodic acid solution, mixtures of the two solatione were. made as shown in Table I. IABLE I mm 0! PERIODIC AND OF IODIO AGES Gannon gtion Concentr tion 10. of 0.000]. M Solution _. 3 10" x 10' w w o 0 Ac d c T 9 1 15.00 5.00 8 2 00.00 10.00 7 3 35.00 13.00 6 b 30.00 20.00 S 5 25.00 25.00 h 6 20.00 30.00 3 1 15.00 35.00 2 8 10.00 1.0.00 1 9 5.00 16.00 if the experimentally detenined optical densities are foand in i‘abIeIIandacaIpas-isonoftheabsorptionccrresofo.0001Hperiodic acidend0.00011|!iodioaoidarefoandinriml. 28 fiflnaeogogeoeufiofloguogfléoaooge 000.0 000.0 20.0 000.0 03.0 000 R00 000.0 000.0 02.0 000.0 00.00 «3.0 000.0 0.8.0 «00.0 0.0m 000.0 nude 03.0 000.0 ~m~.0 000.0 000.0 03.0 80.0 000.0 000 03.0 03.0 30.0 80.0 000.0 000.0 03.0 20.0 000.0 000.0 mmm #00 80.0 «00.0 000.0 02.0 000.0 2.00 000.0 000.0 000.0 000 03.0 30.0 000.0 3.0 000.0 .800 000.0 000.0 20.0 00.0. 00.0 03.0 80.0 000.0 09.0 0.00.0 000.0 000.0 000.0 30.0 004 0.000 000.0 00.0.0 2.0.0 0.3.0 nnmd 03.0 m00.0 $0.0 000.0 .34 000 30.0 «and 03.0 03.0 ano ommé 30.0 and 000.0 030.0 000.0 EN 30.0 000.0 H00 03.0 03.0 000.0 03.0 03.0 000.0 30.0 30.0 00.. 02.0 03.0 0.3.0 000.0 05.0 03.0 03.0 0000 0.04.0 0000 08.0 new mum. o «3. o 03. o 03. 0 m3. 0 0.3. o 43.0 8:6 03.0 «5.0 Smé 08 Mod“ 001 e.“ .0.“ en ; em 00W .0 mom .0 .00 mm 0363.30.28 32 3000.0 E toomhdggdaggggaggsg Hug I.oo- <9 0.60 OPTICAL C DENSITIES 0.2m- _ o\o 1 1 _1 .L 220 no 300 WAVE LENGTH IN MILLIMICRONS Figure 1. Absorption Spectra of 0.0001 M Periodic Acid and 0.0001 M Iodic Acid. 29 ‘QEEEEEI orgpoasible errors. From the measured optical densities of the original 1 z lO’hJM periodic and iodic acids the optical density of etch mixture at each wave length was calculated. These calculated optical densities were campared with the experimental optical densities and the deviation noted. This deviation was labelled plan if the observed optical density was higccr than the calculated and minus if it was lower than the calculated optical density. 1 suxmary of these deviations is given in Table III. TABLE III SUMMARI OF READING ERRGRS Vm- Concentrations of Periodic Acid x 10’5 Sum of the length 9 8 T 6 F 11 3 {TI-T- ' Absolute 1M w - m- c 200 +12 0 +6 -2 +13 +12 ~12 -3 ~17 77 205 +9 *6 4:5 4.8 +10 +11. ab 0 --11 1147 210 +7 +10 +13 #36 +10 +18 +1 +5 - 5 105 215 +7 +1 413 +9 + 3 41).; +2 +1; -6 65 220 +8 +6 +11 +9 +h +7 +2 +6 -5 58 222.5 +1. +6 +7 +6 +5 +3 0 +1. 4. 39 225 «+20 +6 +13 «+18 +16 +8 +11 +7 -1 105 230 +8 +11 *7 +8 +7 +5 45 +5 --3 S9 235 +8 +8 +3 +3 +5 +1. +5 +1. 0 to 21.0 +8 45 *8 +6 +6 +6 +3 +3 ~1 1.6 250 +5 eh +5 +3 +5 +1. +1. +2 -1 33 31 Examination of the sums of the absolute errors indicates that tho two lmst sums are at 250 mp. and 222.5 ml. The former wavelength In! the disadvantage of extremely small increments between the various dilutions. The latter has much larger increments and therefore a greater degree of accuracy. This illustrates the principle that measurements made at absorption peaks usually produce the greatest accuracy. From the optical densities of the original 3. x 10"h periodic lucid and 100110 acid concentrations, values were calculated for each of the mixtures. mesa concentrations are canpared with the experimentally determined concentration: calculated from the optical densities of the mixtures in Table IV. TABLE IV ERRORS IN CALCULATING COMMAHONS OI PERIODIC 103:0 Wncentrat ions amoentratim m omted Per Tint W in Moles/Liter V from Optical Densities Error _ 9.00 x 10-5 9.05 x 10-5 +0.55 8.00 x 10'5 8.06 x 10.5 +0.75 7.00 x 10.5 7.08 x 3.0“5 +1.11; 6.00 x 10’S 6.07 x 10"5 +1.17 5.00 x .10"5 5.06 x 10‘5 +1.20 h.00 x 10"5 3.92 x 10‘5 -2.0 3.00 x 10‘5 3.00 x 10'5 0.0 2.00 x 10.5 2.014 x 10'"5 +2.0 1.00 x 10‘5 0.95 1: 10's 6.0 r7 32 Itwfllbcueenfrmhbloflthatforfllexoeptthevarylow cmumotperiodio acid the unmodi- goodtomermofplun ”ma. » mun or the Man method of determination to LE 1 QO’h H coding 2mm solution. A 1.75 x 10"h lulu caution of Wm periodato was made b clinching 3.990 g. (11.5 minimal“) 0: Medic acid in 15 ll. of “to: in I 100-1111. wlmtrio flak, adding I 79.36 ll. of 0.2309 l M Iolntion and diluting to the indicated "1... he uniting ”hum m 0.175 K with "upset to rack (or mung. n11: palatial: m then diluted to 3 molar“, of 1.?5:J.O"h. nmfmmtmmmmmmdmm of “in: to obtain a lolufion and even than it did not cuplotely . dissolve. met-arm, the relatively Balms periodic 0.014 It: used. A 1.75 x 3.0"h H Dilution of Indian iodate 0:103)!“ nude h din-01m Codi“ yiodtto in later. WI of than M W16!!! m we and their m den-tun MW at 222.5 m and 20° c Instr-numb 33 ILBLE‘V 0mm mm or 300mm PERIODHE-SODIUM 10mm mm m 222.5 up my 20° 0. concentration of Ratio by Hole Percent ' ' v v 701m of of Solute Min Sodium Periodate which is ggtical Demitigg Peri Ioda to Iodate Sodium x 10 x 1 Solutioge Periodate Obeerved Calcglaied 1.75 0.00 ' 100 1.73 1.10 0.35 , in]. 80 1.02 1.10 1.05 0.70 3:2 60 > 1.113 1.13 0.70 1.05 2:3 10 0.830 0.833 0.35 1.140 1.1. 20 0.536 0.538 0.00 f . 1.75 . ‘ 0 0.21.2 v..— f w w w.— Efreot of sodium 0122303500 ca 20222 tion of Begodio acid. 'lbe ultra-#1016 absorption curve for a 0.0002 M solution or periodic acid was cmtmcted by measuring the optical density at various malengtha. 1'0 determine the Warsaw, it my, true aodiu bicarbonate, 1.000 g. 0! Iodine bicarbonate was dissolved in 1:85 :01. 0: the above 0.002Haeletimandthaabam'ptiummdetomnedagain. m unite are chain in time 2. It in apparent that the ultra-violet sawed otpariodio acidanaluiacannotbemd onreactionnixturea htfarad with Iodiu Incubate. the carbonate group appeal-a to dim the Win peak toward tho darker wavelengths. It Ia {and a email. maaim that a periodic acid aolatim, ‘Ihichvaah-eated withanequalmberei‘moleaefaodimm'drmde OPTICAL DENSITY r—ox 2.o_ o\° 1 l I L 1 200 220 240' 2—00; WAVELENGTH m MILLIMIORONS E“igllr‘e 2. Absorption Spectra of' 0.0002 M Perioflic Acid With and '.'.'"Lthout Sodium Encarta-late. 3 .‘J 35 wethaamahau'ptimaaaaolationoftheamnolarivuthmt .0 ”din Ivdroude. w. rear periodate determination during the prom“ of a reaction, aliquot emplee were removed at intervala, diluted to 0.0001 H (hued on the annual concentration of periodic acid in the reaction mixture) and the optical demity was read at the mature indicated in the appropriate table . Oxidation of Q-Gmcoae Raactioa or 0.025H1eriodio acid with 0. 45 H Ewan. Equalvoluaaotao.0§0nperiodio acid ooletieuand (10.00901! gig-Macintionmplacediazoom. erlemeyerflaahandatthe talent of initiation of the Men, he aolatiom were rapidly poured mthmthau Bupleanratabenoataarapidhaa We and altemteh‘ toe- periodio acid We and for romalde- Ivan-3311!”- mwmmmemperrmdwmlmu. aliquot cal“ 01 the "mm and diluting to 250 all. 1110 Optical osmium thenaeaaared at 25° 0. and 222.511}. wmmamreaadeh'themethodotfipeokaod tori“ (11). Wt” aliqaotawereruovedlroathe reaction data-MaddedtoalO-al.volmetricnaakcontainioglal.ot o.hleedieeu1nte. Meandflntedtolo.0a1. Otthiareaelting «anion, 1.00 .1. can mam added to a SO-nl. clan-“append We flask. rethiauaaddedoJ m1. 0:105 ohmotropic acid 36 andSml. oflhflenlforic acid. memixtm‘cwaeheatedinboiling eater for 30 minutes. It was then cooled, diluted almost to volume, cooled again, diluted to value and then aerated for 30 minted. he optical density was measured at 570 up and compared with a previously contracted calibration curve prepared from lotionsl Baron of Standards g-glucoee according to directione awn by Speck and Met. Subsequent chronotropic acid analyses for formaldelvde were made in the em m, differing only in the degree of dilution and the quantity of eodiun eulfite need. Then two factore were detemined w the original concentration of the reaction mixture. the reaction Wmmellydilntedtoeuchanexteutaetoehteinbctnen 0.03 and 0.10 millimolee of formaldehyde per milliliter of diluted eolution. me quantity of sodium enlflte wee edJueted in each case to give a eligxt exceee beyond that required to reduce all of the original periodic acid in the aliquot to iodide. In all caeee, a one fillilitcr aliquot of the diluted echticn wee need for the eeheeqnent cola development with chruotropic acid. Table VI ohm the reunite of the reaction mixture analyue and Figaro 3 illustrates the propeee o: the reaction gephicelly. Another reaction we run in the em way as the preceding, except that periodic acid analyeee emplea tore reed immediately inetead of being alloaed to etand for a time. the units of thie analyeie are roundinreblevnendinrignreh. Acomperieonofthetwog'aphe ”idea definite indication of a continuation or the reaction after dilation. 37 TABLE VI more 1010 0mm m momma 300000110 0! m 20102100 01- 0.025 n 201110010 1010 100 0.0015 M 04100032 11- 25" 0. Original Quantities of Reactontu Periodic Acid, 5 .00 11111130ch Q-Glucoee, 0.900 Millimlee w—__~ v— w—t Fmeldegde Produced e In 163 of Periodic Acid Consumed 0 O 68 in mime” Density of Periodic in Hinutee Formaldehyde , Acid W M h'odncedfl 1.5 0.792 1.ho 2.2 0.081 1.25 0.698 1.95 6.7 0.106 7.0 0.670 2.12 8.3 0.110 10.0 0.635 2.32 13.5 0.120 20.2 0.555 2.76 21.7 0.153 31.0 0.500 3.06 33.0 0.160 10.0 0.1475 3.23 1.2.6 0.161 55.2 0.h3h 3.50 57.5 0.173 87.0 0.399 3.71 90.0 0.193 119.0 0.391. 3.71: 122.0 0.220 181.0 0.1400 3.70 180.0 0.270 310.0 0.31.9 3.98 315.0 0.360 w.— Y" editor the periodio acid emples were diluted they were allowed to stand for m- 30 mixmtee to 2% hours before they were read, on the aeemption that me extreme dilution had effective]: deceleretcd the reaction. the anus were then read on the spectrophotometer at the first convenient time. It no later found that there was etill a reasonany rapid reaction taking place in the diluted re- action nixturo. Therefore, the tine interval ie actually longer than the one indicated in the table. men eolsr quantities are calculated for the entire reaction mixture. con .2 r; A ll 0.? pm mmoors. ) : mnog.c cad «Ho: owflOfipen m mum 'C In? Kfi me32_2 . 0 MC coauowom ..P\ REE Om 0 .ml. 0.39% _ zo.»ez=mz L p of of q . m o $|\0\q .. d _ p ~-4 P 4 _ whdoo .mum mun—01.1.4.2 mo>IMOJxw0442m00 wuaoz_J.__2 mmJOZsz The original 0.250 M periodic acid which had stood as long as the reaction mixture was diluted to 0.0001 M concentration and measured. It showed a decrease in periodic acid concentration which corresponded ts 0.28 mini-ole and mich partially accounts for the 0.149 nillimoles excess of periodic acid consumed, inasmuch as 11.5 millimoles represents the theoretical ccnsmuption of periodic acid. 'me theoretical pro- duction of fomaldelvde was 0.9 nillinoles. Therefore, the theoretical quantity of Imaldehyde was not produced, althougx a sligrt mercen- ssnpticn' of periodic acid had ensued. Reaction of 0.0875 )1 sodim periodate and 0.0125 M D-glucose. A reaction was run in which the same concentrations or sodium periodate and Q-glncose were used as in the unhuffered reactions run is Hughes and seven (10), that is, 0.0875 M sodium periodate and 0.0125 n W. Equal quantities 01' the two solutions with twice the above molarities were quickly mixed and aliquot smples withdrawn at inter- vale for periodate end romaldehyde analyses as before. The results cttherescticnsreahalniu‘rableVIIIandinFimresSandé. In interesting observatia was made during the course of this re- acticn. than sodium periods.“ was used in the oxidation of Q—glncose end the aliquots were diluted for optical density measurements, the diluted couples of reaction mixture reacted faster then the undiluted hitters. mat is, diluted suples which had stood for acne time had a lover optical dcuity than suples which were freshly taken and diluted. With the periodic acid reaction, Just the opposite was true. he examples given in Il'ahle II will illustrate this tendency. he TABLEVIII 222100221: 0mm m) FORHAIMHIDE Penman 3! me morgm or 0.087539001m12m10mnm0.0125112-0100052u20 c Orianal Quantities of Reactantsx Sodima periodate, 8.75 Millimles 1101:0000, 1.25 11111111010: r , _r L I an:_ e I m E; Wmec gficaclnmmiifioles fiePEt—efimw .dBME; 0?- in Seconds Densiw of Periodic in Seconds Fomaldelvde M Acid Unnamed! Produced! 82 1.50 1.30 151. 0.156 21.0 1.38 a. ‘ 2.00 327 0.156 1.91 1.28 2.65 560 0.176 673 1.23 2.95 7111 0.176 8M1 1.18 3.25 912 0.192 1,071. 1.16 3.2.0 1,152 0.190 .. .... - 1,332 0.192 1,630 1.09 3.80 1,703 0.192 2,072 1.07 3.90 2,112. 0.206 3.538 1.01 11.25 3.6011 0.200 1,367 1.00 11.30 11.150 0.203 6.533 0.957 11.55 6,707 0.218 9.326 0.922 11.75 9.1.05 0.222. 13.273 0.903 11.85 13.578 0.218 15,216 0.883 1.95 15.310 0.200 22,133 0.858 5.10 22,237 0.236 26,003 0.81.5 _ 5.15 26,085 0.256 28,580 0.832 0 5.25 28,701 0.280 32.173 0.820 5.30 32.570 0.296 35,010 0.816 5.2.0 35.187 0.312 71.355 0.155 5.70 70.097 0.500 2121.1: VIII (00172120213) .___AA. _1 _._d._ .-..-—.g Periodic Acid Consumed Formald e Produced Wale 00210.1 111131010100 * Wrens erv no ea c Densiw of Periodic Fomaldehyde w w 5010 0000793002 , Produced.» 32 hrs. 17 m. 0.716 5.95 32 hrs. 19 min. 0.662 16 hrs. 30 min. 0.675 6.15 11 hrs. 32 72111. 0.800 56 hrs. 17 min. 0.703 6.00 56 hrs. 18 min. 0.912 67 hrs. 38 min. 0.691 6.10 67 hrs. 35 min. 0.982 79 hrs. 25 min. 0.665 6.25 79 hrs. 30 Iain. 1.012 92 hrs. ' 5 11111. 0.662 6.25 91 hrs. 37 min. 1.07 128 hrs. In min. 0.665 6.25 128 hrs. 10 ain. 1.11 139 hrs. 10 21111. 0.651 6.30 139 hrs. 10 min. 1.11; 189 hrs. 10 10111. 0.653 6.30 189 hrs. 1.16 11 days 1.16 Ilsnk 1.71 mess solar quantities are calculated for the entire reaction We LM .0 com um onoonamum z mmflo.o was mpmoowamm :nfivom z mmao.o mo Cofivowem .m ea:Mfih nv.u my. x” m G 7_nvuu w m ON 0. w. c. N. o. o w e N q _ _ _ a 0 l q q _ l5 7 ||||0 v! o 91 d o C c 10 1 zo_kosooxm mo>1w04<2¢0m Au.N.l. 71.9.nv c.0101 Lee I L 0.0T . _ L- - _- , _ _i-.:- _ _ 1:. g _ gm l w.rd.unv_m_w.u m_Q.>.lm_o.4me‘~unvu mmanvS._Jnd_$_ www.4nvs..4.u.s_ < r d V 1} UN!“ \Uuz .0 com um oneoSHolm. z mmfiod 5A.: mpmvowhmm 1.5.300 E mwmqo mo soapowmm .0 mafia 1.5 m><0 __ O. m m N .0 m .V M N _ _ _ _ _ _ _ _ _ s . O .1. N.O l¢0 ON]. O.ml .100 O .1 1 o ORVI m 0.0.! % O. .l ZO.PODQOmn_ MQ>INQJ:.uo..<2 mo... mmsoz.44.z mmnoz_4iz ILBLE IX 116 COHRIBISOI 0P REACTION RATES BEFORE.NKD.AFTER.DILUTION Sodium PeriodateWReaction ample taken at 6533 9000. 0 hr. (380 Tfible VIII) and ibich stood diluted for the 0.957 Wed We Sulples taken after 6533 seconds 9 the specified tine and freshly diluted. ”1‘ wt“ ‘t 32 M's 17 “line 0 hrs and ibidh ltOOd diluted for “I. Mined times 0.716 Suple taken after 32 hrs. 17 min. 0 the specified tine ad freshly dilated. gericdic Acid Reacting 0 hr. ample taken at 1% min. (See Table VI) and which stood dilnted for the 0.792 specified time. 8-991» 2.101111200013011. ethespecified tineand freshJJ'dilatcd. W Cmtical Densitieew 3A hr. 0.880 0 .992 31 min. 0.762 0.698 0101.10. 8.0100010002557010. mduhiohstooddilltod M the specified time. 0.11311 ample tekonafterS’Snin. e the specified time and freshly diluted [4% hrs. 2% hrs. 2‘]. hrs. 0.785 0.750 0.680 0.858 0.806 0.755 0.670 0.000 18 min. 0.1127 0.399 h? Oxidation of Q-Arabinoee me reaction was conducted at the following concentrations: 0.0200 molar in periodic acid and 0.00145 molar in Q-arabinose. 1'0 start the reaction the two solutions, each twice the above concen- tration, were quickly mixed in equal volumes. Samples were removed as before for periodic acid and fmoldehyde analyses. .. Periodic scid enables: Ono-milliliter aliquots were taken and dilated to 200 m1. weaves analyses: Two-milliliter aliquots were moved and addodtooncmillilitcrofO$Hsodiueantemdthodwleto 10.00 :1. 0! this solution, 1.00-m1. aliquots were taken for the Mopic soid deternimtion as given previously. mommuorthe mineamgiveninhblexminrignre 7. 118 IABLE I ”3101110 ACID 0013mm ADD ME HMO!!!) I! THE OTIG 0? 0.0200 1! PMCDIO ACID AID 0.00115 )1 13-131mm A! 0. Originel Quantities of Reeotentst Periodic acid, 11.0 Minimise g-Arebinose, 0.9 Hillimolos Periodic Acid Oousuned Poneld¥e Produced 11 en 0 es in Seconds Density of Periodic ‘ in Seconds Pomeldelvde Acid Consumed. Prodoxggd‘l 66 0.680 1.67 109 0.056 170 0.587 2.08 226 0.056 318 0.538 2.29 386 0.0614 1:73 0.1185 2.53 560 0.052 653 0.h61 2.66 712 0.072 816 0.151 2.71 882 0.076 1,220 0.113 2.80 1,286 0.080 1,8m 0.1125 2.811 1,872 0.092 2550 0.1116 2.88 2,631 0.116 3,550 0.109 2.92 3,650 0.11.6 5,200 0.395 2.99 5.300 0.182 7,100 0.373 3.10 7,500 0.220 8,200 0.370 3.11 8,300 0.236 10 ,107 0 .357 3.18 10,200 0.252 15,655 0.328 3.31 15.765 0.3814 19.1155 0.309 3.39 19,850 0.1156 22,700 0.29? 3.111. 22,800 04:92 25,850 0.287 3.118 26,000 0.512 39.000 0.278 3.53 29.150 0-572 30,850 0.271 3.56 30,932 0,591; 10,000(20 1111.10.22? 3.11: 72,000 0.828 14 am 00215 3083 h d”. 008,10 31ml: 1.01 'nzese mole: qnentitiee ere celmleted for the entire reaction We 119 .0 0mm aw mnognw: 930.0 05 «40¢ oavoahom 2 080.0 mo coapowom é. ohdwfim 000. x movzoomm ON 6. 6. e. N. o. o m c N e.__,___n__q__ie___d_ $.01. 0.3: .v.«1 ~61 zo. . .. m m _ F — p — . _ _ — b p — _ _ _ _ p 0.260;: . motimgezgd mmnozfinzz mmnDZSiz 50 Reactions of Periodic Acid with Various Gaupo‘mds thich are Not Sugars oleld e. i check on the purity of the canpcnnd w periodate oxidatim ad fmaldehyde analysis, using the method originated by Spock and Forist (11) showed it to be about 90% pure. Periodic ecid in 0.0050 H concentration was allowed to react with Wdetqde in 0.00145 K concentration. This was done 1v rapidly urine equal volume of 0.0100 H periodic ecid (2.0 nillinoles) end 0.0090 1! glycoleldehyde (1.8 nillinoles) eohrtions. Analyses were made 17 the sue nethode need in the preceding reaction. the first periodate euple removed (after 121 seconds) indicated e periodic ecid consumption of 1.81 nillimoles end the first Imelda- hyde melysis (189 seconds) indicated 1.58 millimoles of fomaldehyde. he funldeiwde production did not increase with ties to any sppreci- sble extent. his production of fmaldelvde no 881 of theory, Ihichcalpsredwith apnrity ofehcrt90$indicatesereeotion practicelly complete in 189 seconds or less. Even if this compound were sveilahleinepnre fonitwosldbequite difficult tomeesure with respect to reaction rates. Its rate of oxidation must be extremely repid. More . A reaction between glycerol and periodic acid gen results very mm to the glycclaldelvde. 'lhe solstice st the beginning of the rustic: wee 0.555 M with respect to periodic acid endO.250Hwithreepecttc glycerol. me reactionwasmnat 0° 0 and the periodete wee titrated riths tended sodiu thioqufate solution. 51 Moldehrde suples were run, as previously described, h the chrono- trcpie said method. me results of the reaction ere given in ”I. 11. TABLE II PERIODIC.ACID censuses eon 102811023102 PRODUCED a! one nencrxoa or 0.5551mooxcscmno0.250xmmno°c. Original Qoutities of heutente: Periodic mini, 16. 67 wholes 0]:cerol,7 7.50 Millinoles m 1+ W e :2: Periodic Acid Consumed Fomsldegrde Produced fine Hilliaoles M cent 0 ‘ MiLLiJeoTes Per cent Intervals Periodic of Intervals Periodic of in Seconds Acid Geno mom-etc» in Seconds Acid Con- necrot- snmedfi - ieel A 1 gamed: icel 278 11.6 97.8 101 111.1 93.7 1,288 111.9 99.11 1,186 114.8 98.1; 2,101 15.1 101.0 2,301; 1h.8 98.1. 130 further incense wee observed beyond these who AMWA. nose noler onentities are calculated for the entire reaction mm s mycerol also shows a very rapid reaction with periodic acid, too repid fm' kinetic determination by the usual titration methods . Another trul using a solution 0.0100 11 with respect to periodic said and 0.00175 14 with respect to glycerol, but otherwise under the sac conditions, gave the results shown in Table III. 52 mm: mm 1010 mum m mm PRODUCED 8! m mom: or 0.0100 x PERIODIC 10m m 0.0th M QIICEROL 17 0° 0. Original Quentities of Reectmtsu Periodic acid, 2.00 Millimoles Glycerol, 0.900 Millinoles I r. 1.111. ‘V .__w._. Periodic Acid Consumed Fomsldglgde Produced 0 Mil es r ce Intervals Periodic” no? Intervals Periodic of in Seconds Acid mooret- in Seconds Acid necrot- Consumed! icel fl Consumede icel_, 187 1.08 60.2 96 1.15 63.7 996 1.69 98.0 8M4 1.6!. 91.0 1,775 1.75 97.2 1,661 1.76 97.8 2,522 1.78 98.8 2,609 1.76 97.8 3,517 1.77 ' 98.5 3,381 1.76 97.8 7,708 1.78 99.0 7,522 1.76 97.8 22,037 1.78 99.0 21,757 1.71. 96.6 ’31:” noler quantities ere calculated for the entire reection nixture. ' Itisddifionlttoeeehw the Imeldehydeprodlcedmdths psedodioecidcmedcsndifieretthssaetinsintervel. Meta-e theseditterencesnsthedsstoenelflicelerms. Adcpli- estsrmottheseeereestieosersshilu-reselts. Meeppeerstoheeslusrreectionintheseredihteseletion es caper-ed with the sore consentrsted solution. WMQE’L mthsehsnssthetpsricdiceoddliflxttore eeuplexsithsaneoqolndsdxiohitooeldnotclem, enuhsref 53 likely cmds hering 1.3mm groups were cuhined with periodic uidineolutimmwe reactionmirbureexnincd inthe spectrophotuster for “gas in absorption, either or the periodic eeideheorpticnerferthe eppeerencs ofnewhends. i technioel credo of trinetlvlene glycol wee used end so: propylene or ethlene glycols were destroyed by trement with sodiu periodete salution. ‘l'he excess periodate wee reduced by eodiun lultite end the resulting solution wee pessed throng: e Dower 50-121 (100-200 mesh) entice enchenge column which wee oherged with hydroniun ions. The solution wee next peesed than e Down: a 10 (50-100 mesh) eniee exchange colusn charged with carbonate ions. no solution was peseed through repeetedlyentil it "longer gees eteet for iodide ion end until it wee neetrel. the weter end other low boiling imities were distilled or: st 117-120 a... pressure end finelly the methylene man was distilled over st 115~119° c. Fifty milliliters of s 0.2 M solution or triesthglme glycol wee nixed with e einilu- velus of 0.! I! periodic ecid solution. duple- ef the resulting solution, 0.1 I! with respect to eech cuponent were stained for possible evidence of e complex es were dilutions of the originel mixture to concentrstions of 0.01 H, 0.001 I! end 0.0001 n. i foresldehyde deteninstion wee pertomed on the reaction mixture. About 0.1 nillinole of foneldatwde wee found to be present. his evidently originsted from some Md 1,2-zlycol. n1. optics). density of the 0.0001 1‘! solution st 222.5 mu indicates the diseppeerenoe oi‘ 0.6 nillinoles of periodic said. this would leewe e net or 0.5 nillinclee of periodic sci-d uneoommtei for. this is each s smell frection of the total that inseam: no no absorption winch could be due to I couple: was found it m be acme-.1 that Mostlylm 313061 does not fem e coupled: with periodic acid and the disappearme of thil mimic ecid cmld hem been due to en immrity in the glycol. Amtmglamstom. this cmonnd Hes exeninei in the some way as the Mnefln'lene glycol. After otmding for four hours the Mixture still had its original content of periodic said end no umseel eheorp- tion peeks were found in the mac 200-350 mp. ”mil Acetometntg. this else we no evidence of my decrease inperioietewhenchcckedinthe mew. ice-3.11 acetone. lain equal qusntitiee of 0.2 H sooty]. acetate endOJHpex-iodio eeideolntiunwerenixed, theeeskingthe result- ‘mmoannmmputtomhomponmt. Attorellmdn‘the nixtmteetmdtwohmetmtmperstm-e, thmdilntingone lilliliter of the mixture to one liter and checking its ebeorption in' helmet itwesfound thstehmtonehslrottheperiodie e013 Ind been reruced. Upon measuring the eheorpticn cone of e corne- pacing 0.0001 F! ecetyl ecetcne solution, it was found that in the motion mixture the ecetyl acetone absorption hed eleo decreased. loom ecetono hes no absorption peel: ct shout 2'73 :91. the rotation mmellowedtoetondfcretoteloffihm stehich tine the periodic said had quantitatively disspmmi. In feet, the Witch Use slid'ntly less than M would he expected from the remitting iodic acid. ittheend offlhourleboutoneheli‘cfthe scetyl acetone sheorption, es mounted at 273 no, he: .Iisappe 211883. the msulte of the resction ere mom in Piano 5. mmforo, there definitely some to be e reaction between 9.09an scotone and periodic said. The entire reaction was repeated using I! 0.2 H sodium pcrioriete solution metre?! or periodic acid. A sheilsr Fraction emefp‘h that it was ouch slow-r, was observed. 258 results or this reactim 216:; be seen in Figure 9. After sequins 2151 been more: for optical density meow“ st the max of 31m :27 harm, the reaction mixture on mama. to steal in the dark at roan tewareture. After 270 cm s smell any Moll-yellow precipitate was observed in the fleck. 1t wee filtered out, fleshed with water ens! dried. It had the do: of ioiofm, hit in melting point wee rum to be 113° c an very can: thus .mpmvmg em moicion of its being iolofons (ELF. 11:?” C). i mastoid test indicated the press-shoe of e hslogen. The empouni may have been twin-ionic acetic said (ELF. 150° C) (72), elthmzh st the time this possibility wee not meted an". therefore the coupoond was not identified. in stmnpt wen we to determine the stoichiametry of the reaction between acetyl sector-m end periodic acid by the titration of samples free e reaction nix’mre 1/60 2.: with respect to noctyl acetone and 5/68 M with moroct to periodic soil. Smoke of the reaction mixture were added to s solution containing excess pctncninn iodide, sodium “outcasts end e measured qumtity of standard coiium menite solution. 56 .cadmsona m on 0:0 umpafifin “asepmua thmo< z H.o and “Ho: owvoapom z H.o mo ohspxfis coapoamm man no «upoonm :oapmnomn< .m wh=Mfim m20¢o_2_44.z 2.:pczm4m><3 r. .2 Hooo.o ocoowod Hhvwo< An“ was: mm .893 9.3pr 838mm 5 mason m nmumm mpszwz :oapommm ANV : Hooo.o vac: caveapwm Adv I . OIO\O lo — >».mzmo 4h_mzuo 30:3 63 1319 second trial gave the following results: Time Interval Holes of Periodete for Mole of Dimedone 1 day 1.436 2 days 1:31 6 days 5.00 The sixth day the reaction was brown and contained iodine. This r"— iodine was filtered off and the remaining iodatcs and periodates were reduced by bubbling sulfur dioxide through the solution. Sodium bicarbonate was used to partially neutralize the acids in the reaction mixture in order to obtain s more cmnplete reduction. The solution , ,j was then concentrated in a vocum and extracted with small portions of other. no other was evaporated and a mall quantity of crystals appeared. These were recrystallised twice frm three drops of distilled otter esoh time. The melting point of the crystals was found to be 101-102“ 0. This corresponds reasonably well with the melting point for 3 ,Bdmethyl glutario acid shioh is given by Heilbron (73) as 103-401." 0. In a subsequent preparative attaupt, similar crystals were obtained free a solution 0.025 I! in dimedone and 0.117 M in periodic acid and which was allowed to react 95 hours before filtering off the iodine and extracting with other. In this case the iodine was accompanied w a terry organic substance thich lodged in the Mel. his time 0.730 g. of crude crystals were obtained frw 600 ml. of reaction mix-t tore, which mama]: had contained 2.100 g. of dimedone. After these were recrystallised frm water the melting point was found to he 99-100“ 6. 6h m acetyl mtcnc. A solution which was 0.2 H with respect to periodic acid and 0.0h H with respect to methyl acctyl acetme was allowed to react at roan tenet-store and the periodic acid consuption was determined it sodium thiosulfate titration. The periodic acid ccnsmnption was found to be as follows: Holes of Periodate Per Hole of {‘3 Time Interval Metlyrj. Acetyl Acetone LE 6 17*} hours 1.5 I 1 1L5 B/h hours 1.6 6 days 2.06 «1' It is evident that this reaction neither proceeds as fact nor consume as much periodatc as the reaction with unsubstituted acetyl acetone. Iodine crystals were noticed in the reaction mixture after 25 due. Phenyl acetyl acetophenone (C(éHSCHzOOCHszsHS). :33 augment, absorption spectrum of this cmpcund was determined as a 0.00001 )1 solution in methyl alcohol as a solvent. See Finn ll. Absorption of this diketone was too mat in the region of 222.5 an to pemit use of the spectrophotanetrio method of determination of periodats. he relative insolubility of phswl acetyl acetophenone makes a quanti- tative reaction in aqueous solution very difficult. To produce a reaction, 5 :1. of 0.2 M phenyl acetyl acetophcnone in methyl alcohol solution was mixed with 5 ml. of OJ; M periodic acid. Upon contact with the water the phenol acetwl acetophenonc hmodiatcly began to precipitate. then the flask was shaken, large bubbles of gas ‘ cane frm the solution. me flask was put on the shaker for 1’; hours .é'sd .eaocogoopood. H503 @805 z muoH x H Mo rafipooqm. nogaomne :3“ egg mzomo_2_.._.:2 2.... ozmqu><3 0mm own Don L. owN CON J /.\/O _ _ _ _ _ _ _ >._._mzwo 440:1...0 67 acre WV to react than in the portion having a loser concen- trstiocotpes-iodio acid. To overcome this nixingerror aPyrexnixing tabesasssdeinshichthemmeonneotedWagx-ound glass Joint, were we at an angle of about 120° to each other. It was then possible to cool the solutions separately in each arm and at the moment at sizing quickly turn the tube so that the two solutions would both m... be rapid}: contained. w quick]: turning it back and forth several f tiles, sizing was done very efficiently. :lhe second error appearing in reactions utilising less than theo- retiesl “to of periodic acid, vas cased 1w unreacted sheet-aldehyde. -. J fieretheresas I! appreciable ascent of moot-aldehyde renainingnn- eons-ed, a hrosn oolor appeared in the chrmotropio “1a detemination of toualdelvde. Ionally a reddishwiolet color is produced in this detaaiaation. the uhstanoe producing the bran color also exhibited fluorescence. msphmenonwasrepcrtedlvmsrntmandsmck (fit) as a product of the reaction of sulfuric acid sith glyceraldéhyde. n had also mean been reported by Eegrile (7S). 1's means this error, the nethod developed w Rash (76) for the W of terasldehyde sas checked in the presence of glycer- Wuflddmwfmdtohemefmmmew mesa W. the method was standardised Iv means or resumed WWuflusm crammqumtityottmaldec- m, assoc-Iced osthyhaohdyen (77). The standardisationonrre was. caste-acted tr. dz point detersinations, which produceda atraidxt line. 68 me procedure used to detersine formaldehyde by the Nash method no as follows: Aliquot ”spice of the reaction mixture containing no excess periodate were dilated until the solution contained frcn one to four sicrogrns of formaldehyde per milliliter. Five milliliters of this dilltesolutionsereaixedsitthl. ofthe reagentsnddlmdto stand for 5 hours at roost tmerstnre. me optical density was then seassx'ed at hl2apagainst ablankaadeupatthe ascetineasthe mile mi made by taking S :1. of the reagent and 5 ll. of water. , m] .i'.-’ Chm'uf I: he observed optical density was coupes-ed with the standard curve and g i the concentration of formaldehyde was calculated. I- H In order that the reactions or glyceraldehyde might be better understood, its depolymerisation reacfion was smdied. he crystalline gyms-looms mm as a diner. mm (78) lists the molecular seigit of glyceraldehyde as 16!; one hour after dissolving the crystals, aleQafteronedqandasthe oalculatednolecularseidrt 0:90!” themai'tertwodqs atremtmperature, bathe failedte give the concentration of the solstice. Diueriaation, such as this, could have an appreciable effect on the reaction with periodic acid. Meters, the ecqaeriuental molecular weight and the equililnviun eon- m were found at 0° 0 by the method or change of freezing point. Much as glyceraldel‘vde is very difficult to dissolve it sea not found practical to follov the course of depolynu'iaation with a rate study. 1 with” of 0.2550 g. Qénghceraldekwde in 15.00 ll. of carbon dioxide-bee om distilled water was placed so. the shaker 69 at roan temperature for about one hour until all the crystals were dissolved and then cooled to 0° C. This was then a 0.3 nolal solution. lhe freeaing point was measured with a Becknan thermometer and compared with the freesinz point of the water. In two trials the equilibrium constant for the depolymerisation reaction was calculated at 1.03 and 0.99 respectivelar and the calculated pm. percentage of diner at equilibrim was 30.9% and 31.6$ respectively. F- In the second case equilibrium was reached in 2 hours and 2S nimxtes F after mixing, but in the first case a somewhat longer time was required. F B: the use of this equilibrium comtant a 0.1 H ghoeraldehyde F solution at equilibrium at 0° c would have approndnate]: 16% of the solute in the form of the diner. Three series of glyceraldehyde oxidations with an excess of periodic acid or sodim periodate, were run at 0° 0. Table II shows the effect of a change of concentration on the reaction. In this case the ratio of periodic acid to glyceraldehyde was kept constant at 10 a9 , that is , 10$ of the periodic acid was in excess above that which was theoretically required for oxidation of the glyceraldchyds. It is evident from the table that an increase in concentration produces a decrease in rate during the first stage of the reaction and an increase during the last stage. Table III shows the effect of concentration on the reaction of sodim periodate on glycol-aldehyde at 0° 0, still maintaining the same ratio of periodate concentration to glyceraldehyde concentration. Again an increase in concmtration produced a decrease in the rate of lumen EFFECT OF CONCENTRATION ON THE REACTION 01" PERIODIC LCD) m WE 11' 0° 0 a. ll_,l ,l. a===:§ 22.22 Millimoles 0.555 M Periodic Acid __ 10 I-Iillimolos 0.250 M Glycergdemm fierimiio Acid filimtptionfl Formallieh do Seduction ‘ Mtervfl Milii- Per cent ime Interv er cent in Seconds Wmoleee necrotical# in Seconds noleen Theoretical 220 1h.37 11.8 103 6.68 68.8 1,378 15.22 76.1 1,271 7.30 73.0 2,525 15.10 77.1 , 2.1.3? 7.1.1. not. 8,092 16.70 83.5 7,976 7.70 77.0 15,052 11.1.? 81.1; 114,937 8.22 82.2 21,910 18.hh 92.2 21,780 8.71; 87.1. 25.202 18.247 92.3 25,080 9.02 90.2 22% hrs. 19.86 99.3 22% hrs. 9.56 95.6 4.... “none molar quantities are calculated m the entire reaction mixture. { . ._.__._«-. _- . w . . o 1 fl . . . . , M ., .l . . . . . . . . . a . i A. r i , r . . . . . n w . . . . n . . . a . .i . 4 . r . , . I v. I r . _ . ._ I . 4 , - I l . w . . , n l... - .\ r _ _ .. . a . a h . . ‘ . .. .2 _ . a . .s O O O _ , . . A, . . _ . . F. n O a e u a . . _ . M _ _ . . l ; n. . . _.. . . a I _ _ ” “---*-vi—W~~ "—7 » . _. 0015' Wall!!! 01 ‘ ‘v I a l 1 a I a was III EFFECT (F CGCENTRATION ON THE REACTIG 8F SODIUM MONTE AND OLIC‘EIRAIDPHIDE 11' O C lg gillimoles 0.100gh Sodium Periodate . Nflllinoles 0.0 g Hiogzgeraldegzde ""“'Pér1odatoigggigggglon , __f“' ornaldgggdefiProduction terval - er cent a erv N’ - r cent in Seconds moles! theoretical in Seconds moles! meoretLeL 159 7.06 78.5 93 3.88 71.h 97h 7.30 79-9 900 3.71 53-5 1,870 7.33 81.5 1,796 3.80 8h.s 2,777 7.25 80.6 2,708 3.90 86.7 8.770 7.35' 81.6 8,685 3.82 88.9 10.597 7.55 83.9 10,890 3.91 86.9 21,065 6.83 75.9 20,911 3.98 88.5 22,788 7.h6 82.9 27.200 7-52 83.6 29.795 3.98 87.6 ‘ihese solar quantities are calculated for the entire reaction.sixtare. . .-.—.—--...— "g... .. . 72 thereactionatthsmmlstaaeandelsointheaoreadvanoedstage. Intact, withnm-ecmsntrrtsdsoluticnsandsodinpsriodateitis difficult to approach theoretical yields. Table IIII illustrates the effect of extra acid (perchlorio acid) or base (sodius hydroxide) oaths reactionuixtu'e 0.100Hwith respect toporiodicacidand0.0hSHwithrespecttoghcec-eldelvde. Itm be seen‘ircnthedata given that anincrease inaciditr decreasedthe rateatthabaanimcfthereaotionsndincroaaadittomdtheafi. file addition of an equivalent number of moles of sodium hydroxide, thatischanangpericdicacidtoscdimperiodate, doesnotsffect the reaction greatly at the initial stage, but does decrease the rate atthelutstagesuntilitisdifficulttoachimatheoretical yield. Inallofthe preceding series involving Wands, the periedatewasdeteminedtvthesodimthiosulfatenethcdmdthe fornaldelvde w the chmtropic acid method. Aeorieswasruninwhich less thanthe theoretical wanton! periodatewasused. Inthisseriesthefonaldehydewasdetenined hills Rash methodusingacetyl acetone, and the formic acidwas deteminedhyfltrationofthe reactionuixturewithadiulhydmxide. his ionic acid deteminaticn is possible in the case more no periodieecidrenainsattheendofthereacticnandthsrensimng iodisacidisdetiutehmvalent. Aliquotsofthereactionnizture Imtitratedfortotalaciditymdthefomioafldmdstmedh «haunting the known equivalents of iodic acid. Tochckthevalidib'eftitrttiufosucacidiathem cficdisasid,steulu'dselatimofeachwereaadeandtitratedia MEXIII EFFECT 01‘ 10mm on 138 mom! or 0.100 M PERIODIC ACID AND 0.01.51! GLICERALUEHIDE 11 0° 0, 10 1111111401123 PERIODIC mm, 8.5 MELLIM0183 GLYCERALDEHYDE 11 09 c Reaction Mixture Contains Perchloric Acid 0.27714 firfodlc 1618 ficmhmad maid dfPi-oduced’“ We: cent HE mama h- Per cent fl'nmee eolar qmtitiee ere oelonhted for the entire meotion mixture. in Seconds moles! Theoretical in Seconds moles! Theoretifi 188 5.29 58.8 100 2.68 59.1 1,188 5.98 66.5 1,070 3.13 69.5 2,171. 6.211 69.11 2,090 3.39 75.3 3,721 6.76 711.0 3,655 3.59 79.8 7,130 7.86 82.9 7,083 3.68 81.8 13,261 7.99 88.7 13,161. 8.11 91.3 20,690 8.31 92.1. 20,600 11.02 89.1. 17 hrs 8.91 98.9 17 hrs 11.52 100.1. ___.-‘ 711 the presence of each other. Mixtures consisting of 10.00 ml. or standard focmio acid and 20.00 ml. of standard iodic acid were mixed and titrated. Two such mixtures were titrated and the volumes of standard sodium hydroxide required to titrate the formic acid in the presence of iodic acid were calculated. The fomic acid in the mix-— ture required 3.96 and 3.91 ml. of base respectively as compared with 3.93 ml. and 3.91. ml. for 10.00 :81. of formic acid solution alone. Therefore, the method appears to be valid for the determination of formic acid. These reactions involving less than theoretical amounts 0: periodate were run at 25° 0. The periodate and the glyceraldehyde solutions were mixed rapidly and allowed to rennin in an saber flask at the specified temperature for two hours before the analyses were made. kl attempt was made to determine glyoxal in a reaction mixture by precipitation with sanicarbazide hydrochloride, but no precipitate of the sparingly soluble disenicarbazone was obtained, even with a deficiency of periodate. Therefore, any glyoxal produced in the reaction must be of such a small quantity so that its eemicerbasone is within its solubility limit in the reaction mixture. Table XIV gives the results of the series of reactions involving less than theoretical mounts of periodic acid. In this table the oriflnal glycereldehyde and periodic acid were detemined kw weight and the formaldehyde and formic acid W the methods previously described. All other quantities were calculated by methods which will be discussed later. 00.0 «2.0 000.0 00...” 009.0 000.0 0.2 0000.0 do 0.0 000.0 000.0 000.0 000.0 .80 0.0... mood do «do 00.0 mood 00.0 00.0 «do 0.0a ”0.0 do 000.0 8.0 no.0 0.0.0 3.0 30.0 o.m No.0 do. ~0d0 00.0 ado 00.0 «m0 ”0.0 04 do do «0d0 00.0 fid 00.0 $0 00.0 04 do do 0030.800 e.“ 33 30330.0 no 330350000 one £390.50 one: 0.“ eubfioaoobnu Mo 8305:0800 nouns 5" union 0.0050305 .0000 800000002 0008.330: 0.2 00.0 0.00.0 3.0. 000.0 40.0 0.0 «.0 do 0.0.0 0.0.0 «0.0 3.0 00.0 00.0 04 d0 d0 mga figsmgmm SEE mmm.o 03.0 H06 mad 035 ~90 0..” H.o H5 $0.0 2.0 H05 mo..o mmH.O Mega o.m 03.0 ~.c . .664» Bi 9.31% EBEHHO 9.5 0.00900th 005980 1.010300 0 0000.0 003080 080080 0000020 00300 .0305 304 00.3.0 as. .3320 055 00000 E00 Junta 0.53.5 $.00...me 330 5053.0 304 0003.00 no 3oz .30 Seem 030% on .980 and cannon—”Ego HG mEQHmhoEdmmmaho mag 5mg 75 . .0020. 103.500.“ 05 5 003.3050 you 030000." 0.3 30.3: no 300 on». .0090 4.40.3003 0.00 0020—. t 0000.0 mood . 000.0 000.0 000.0 «0.0 0.0 00.0 00.0 000.0 8.0 no.0 0.0.0 3.0 30.0 o.m «0.0 do: 00.0 000.0 0.8.0 000.0 000.0 000.0 0.0.0 «0.0 «.0 03.0.» 5 00000018008 no «8090030800 05 03. 055-000 9003 .0 300 3000.02.” No 0035900000 05 noun: 5 00.300 0000 00.0 00.0 00.0 3.0 000.0 0.0 000... 50.0 do Sic 8.0 no.0 ~06 53.0 “3.0 o.m No.0 H5:. 0080 a 000.0 00.0 00.0 00.0 000.0 000.0 0.0 do . «0.0 do 0.0.0.» 3 00 05 03.02 5 830988030 003.053 no 80.80 0.0.0 00.0 0.00.0 00.0 00.0 3.0 o.m 00.0 00.0 000.0 8.0 00.0 00.0 03.0 000.0 0.0 «0.0 do. 0.0.0 00.0 mo.0 00.0 3.0 00.0 0.0 00.0 0.0 #:«gaiggufigggégaamfigg gagfigsonfioguoguoofizgflaflg 76 In order to better understand the glyceraldehyde oxidation a series of reactions was performed on glyoxal, one or the intermediates in glyceraldehydo oxidation. 1 5% mental solution was distilled, the distillate thermally mixed and used as the basis for a series of reactions in a solution 0.0145 R with respect to periodic acid. One solution was made 0.3 I! with perchloric acid, a second was used as it was and the third had an equivalent amount of sodium hydroxide added to it to produca'monoeodimn periodate. no progress of the reaction was followed 13 sodium thioaulfate titration: for periodate oonomned. This change of pH produced no significant differences in the rates of maction. MIC! Q-Glucose It is apparent fro- the results of the caddeticn of glucose that the production ct-femaldehyde does not keep pace with the consumption a: periodsts. m: rm was also reported is speak and roe-1n (11), the obtained practically theoretical consumption of periodste while the {maldelvde production In still less than tve-thirds er the thcoretical Vllue. Ed». and Emil (10) noted that dun the periodsto was determined with ”din thicnlfste there m s realm): clue correlation betaeen the periodste consulted and the tennis acid produced, htuhenthe periodstemdatorlinedlvnemefarsenitethepericdete consumption far exceeds the tonic said production. Indies Ind Revel]. attempted to explain this w see-in tilt the vaults did not reduce the caplex which was toned bottom glucose and periodate and that therefore the arsenite method mlysed only the em periodtte. If a rovereible “amen ensued between on Periodatc + glucose m complex periodatc ad the glucose to tone the cmplex (Whether a not this ample: is an intermediate hetleen the reactants and the products or tether it is unproductive ouplexmakes no difference inthis cum), it would be expected that the couple: would disappear within the 15 minute Iaitdng period when exposed to excess sraenite. Il‘he procedure in this method was to add sodim bdcsrbonete, an excess of 78 standard arssnitc solution and than after 15 minutes to back-titrats the anus of arsenite. ihe arsenite should then 17 reacting with the tree periodate have reversed the reaction, causing the disappearance of the emplax. If the cmplsx is an intermediate in tho reaction and it its disproportionation is vow rapid, as there is good reason to believe, the complex would disappear even more rapidly. mentors, this explanation of the discrepancy does not seen very convincing. his annual: with respect to the arsenite detersination or psriodatc an be explained by the greatly increased rate of psriodats oxidation in sodium bicarbonate buffer as reported by speak and lorist (11). During the brief interval after adding the sodiu hicarhuats and before addins the arsenits the reaction is accelerated to the extent that considerably more periodate has been consaned than was real}: the case at the time of removal of the sslple, uhsreas the formaldehyde or (oxalic acid deteninations which do not involve sodium bicarbonate additions do not show this acceleration. this explanation was advanced hr Van Slyke, Hiller and Machdyen (23) for the oxidation of W lysine. . I! the open chain aldehyde form of glucose were the only form attacked and seaming a random attack upon the bonds, a mole of romaldelvue oagzt to be obtained for each rive moles of glucose. his is not found experimentally by any method or analysis. more an be preferential attacks on particular bonds. within an open chain molecule, hut there is essentially no information on this at present. However, it is not very likely that such preferential attacks would 79 delq the theoretical production of fonuldehyde as 1mg as has been observed, especially in the preseme of excess periodats. It becaae apparent early in the investigation that the periodate attack on the cyclic heniacetal Iona or glncose would yield the {crawl glycereldelnrde as shown in equation (7) and that um new be a relatively stable substance. H- a 2 ROUGH HCDOH 0 11-0-03 4 a i .- 30-0-11 0 3 10h ? n a 0 fl 1 I t ( ) H-c-on c-o o 2 2 Home 7 0 0 I 0 IO n-c n-c-o—c-n """" 34-03 ----"--> 0 11-0-01! 11-0-03 n-c-m I o a HCBO H a a Mdence for this ester has been ably presented In Barker and Smith (62) and by Schopf and Wild (6h). Hydrolysis of the ester would then slowly take place, thus really becoming the rate-determining step. his mechanism fits the experimental facts better than on which has previously been considered. One fact which equation (7) does not explain is the almost imediate appearance of a noteworthy quantity of femaldehyde. his quantity is larger than would be enplained on the basis of the proportion of glucose present in the acyclic form at a given instant and which would then be oxidized directly in that ion. In Minion of this formaldehyde production, it is possible for the equilibrium between the cyclic and acyclic form to continue even 80 after an attack by periodate on the cyclic tom, providing the (ll-C1 bond has not been cleaved. lhis m be illustrated by equation (8). H-G—OH E-G-Q H I \ I I H-C-OH n-o-o 0-0 I i I m ’ 8 30-0-11 o-o ( ) . 4—...) ............> ' H-O-OH l-o-o H I I H--(l--O-J H-C-O-- e I u I ‘ H-c-on H-C-OH 8-0-0 I I I ' H l! B-O-OH -l- I 30003 11-0-03 1 periodate attack on file products formed in equation (8) would then yield the one imadiate quantities or fonaldehyde and formic acid as if the acyclic {on had been tttacked directly. bcperinsntal evidence has been found for this ”chm-Ia. Upon emulation of Figures 3-6 it will be ebeerVed that the initial portionorthereactienis representedby aregioninehioh the slope of the linu ”profiling the rates of pmwete emulation and fonaldehyde production is comparatively great. This changes in the course of the reaction to eventually produce an almost straight line with little slope. me steep initial portion evidently represents the attack on the open chain aldehyde fem or glucose as well as the A stage in equation (7). 81 Ry examination of a typical case such as the one represented w Imam-ad Figurehitwillbenoted thatthe fonaaldetvdeproduotion serve becmes practically a straight line after about 900-1000 seconds. Omsultation of the table indicates 0.185 nillinoles of formaldehyde produced in 972 seconds, which represents the em number of nillinoles of glucose oxidised. Assuming this to be oxidized in the open chain form, this would represent a pericda'tc constnnption of 0.925 sillinoles, seeming a ratio of five to one. Having originally started out with 0.900 nillinoles of Q-glucoee and having oxidised 0.185 nillinoles in the open chain form, there would remain 0.715 nillimoles to be oxidized as the cyclic form. Using the ratio of 3 millimoles of periodate per nillinole of cyclic Q-glucose in the rapid 1 stage of the equation, this would represent the consumption of 2.11:5 millinoles of periodate, er a total consumption of 3.07 nillinolee of periodate for both the open chain fan and stage A of the cyclic form. Table VII indicates that this quantity of periodate was consumed at approadmately the end of 5000 seconds. consulting Figure h it an be seen that frat slidltly bqend this point the periodate constmption cave homes nearly a straigzt line. the slidzt difference batman the calculated point of 3.07 nillimoles and the indefinite point where the change were is very likely due to the slow hydrolysis of the form]. ester which releases more glyceraldehyde to consume periodate. A similar calculation based on the data in Table VIII and Flame 5 indicates a total consumption of 14.05 millimoles of periodic acid for the oxidation of the open chain and the initial stage of the reaction with the cyclic compound. 82 The datainl‘ableVIandthe accmpnvingl‘icheillhudJ: lend itself to this type of analysis because the delq in reading the periodste samples has used the mearame of the periodic acid consumption curve . Q-irabinoee Ihe reaction of Q-arahinoee with periodic acid may be considered on a basis very similar to that of goalucoee, fencing however, a fowl ester or glyccllaldehvde inctaui ct glyceroldehyde. H- 2 HCOOH 30.3.3 . a (9) n—c-cn 0 31° . n-o-og ga-o nag-:11] n~:c-o-c—n rm 9 H- c-ou 3.9;.) m3“ 8-9 . H. a m H e HOOOH the rmaldehyde production curve is very nearly streidlt frm the heximing. berated-e, take the value of 0.056 nillimclee (See Table I) for the first formaldehyde reading, Which is esunned to arise from an oxidation of open chain aldehyde molecules. 'Ihia value would then menu. a corresponding periodate consumption of 0.221; millinolee and “1d suggest that 0.8M; nillimolee of the arabinose would be oxidised in the cyclic ton. his letter reaction would require 2.532 aim-oles of periedate to produce the ester stage or a total pericdete 83 consmption of 2.756 nillimoles. mic is very close to the definite break in the periodate eonsmnpticn curve. (See Figure 7‘). Therefore, it appears very probable that the oxidation of Q-arabinose proceeds by the mechanics indicated, although definite proof would depend on the isolation and characterisation of the formyl ester 0! 317001110 aldehyde. 1 , BeDiketones Oourtoie and Joseph (25) were apparently the first to report the reaction of periodate Iith a 1, 3-diketone in a statement that periodate oxidisee dinedcne (Ll-diasthfl cyclohexan-B,S-dione) (No data on periodate eon-nod or the products formed were given). Very recently Home. md Bobbitt (he) have reported observations we. clearly show that cyclic 1,3-diketcnee in gemral undergo a specific oxidation with periodate , but that this phenomenon does not extend to the acyclic 1,3-diketonee. Although not as elaborate as that of Wolfran and Bobbitt the present investigation has also elucidated the course of the dimedone reaction. Moreover, it has shown, contrary to Holfrcu's report, that acyclic 1,3-diketones (acetyl acetone, 3-nethyl—2,h- pentadione and l,h-dipheny1-1, 3-butadione) undergo oxidation. In general, the reaction of 1,3-diketones proceeds without fomation of tennis acid or formaldehyde. Carbon dioxide is formed and mono- ccr dicarboawlio acids (depending on the structure of the ketone) are the products. lhese facts have led to proposal of the following mechanism for the reaction. 8h 3 0 B O 0 0 0 a a a 9 a a a - (10) R-G-c-C-R' e 1301, R-c-c-c-a' + I03 a -------> I H or the hydrated H 101‘ 3,4106 H I H 0 O 0 O O O a a s e I c (1.1) R‘C‘C'O'R' pd RPG‘G'O'R' I H H H O 0 O O O a a a a e (12) a-c-c-eat n-c-c-on + mm c 2 103' " 21°14. ..__........; 00 as (13) a—c—c—os e 10,: 0 EUR scam oszco3 e 103' is an alternate mechanism for equation (12) the folloeing possibilities m be suggested: H H O O O O O O I I I I I C (11:) R-c-c-c-a 0 IO), , n-c-c.c-av e :03 + non OOO . 00 sea as (15) n-c-c-c-a' + xch" «r ms , a-c-c-os + mm + 103’ ‘lhe latter scheme is that proposed w 'dclfrm and Bobbitt. They suggest oxidation of the enediclone form to the trione and subsequent oxidation of the latter substance, because they observed that oxidation 85 o: mama-.mmmm occurred rapidly with oo-uption or three moles of periodate per mole and also that 1,2,3-cyclchexanetrione rapidlycmed 231a orperiodatepernole. Apperentlythe hate acidisthst‘inalilrtauedisteinthe reactionsincephthslonioaoid thedasitsanflinederivativerruuMdatiend 1,3-1ndendime. meexactnatueotthemflnacidsobtainedmmom 1,3-diketones as not detox-ind in the resent mom no that the queetionoi’vhethuthossseoheni—se-hraoe the oxidatioootsll 1,3—dikotooes main to he settled. Volta-an and Bobbitt included the effect of pl! in their investi- gation, but it mould he pointed out that they conducted their investiutiou oaths effect explwneans “White”. thesehflushsveatendenwtoohsoue the effectorflhthe effect of the phosxhate ion on the reaction. A unnu- effect w occur in oxidation involving the bicarbonate ion. Speoksnd Met (11) found aesthinoreasedrotes inthepses— me e: sodium bicarbonate. Hanover, auctions of periodic acid ehen Wufinequivsleut-ountotsodiuhydroxidedonotsppesr to react in the sue way as in the presence of eodim bicarbonate. hex-immdnm'eZindicatesthatthereissmfiotthe periodste Wimtmi-dlmmlengths asatfefledhthe addition of sodium biccbonate. this niglt indicate a change in the p‘oriodate 1m, which an affect the'rate of the oxidation reaction. Equivalent mounts of sodium hydroxide produce no change in the absorption spectrum of periodic acid. This is a matter that nidmt well be investigated in the future. Wolfram and Bobbitt also detemined periodate w the sodiu bi- oarbonate-arsenite method, mich in the process or this investigation has been shown to give results that are not always dependable in the presence or excess reagent containing active tum-oxen. the iodotorn reaction mixture heme too new competitive with the analytical reaction. literature, the thiosalfate method or the absorption method have both been fwd superior to the areenite method for detaining periodate in reaction sum-es o‘ontaining 1,3-diketeaes. Qé—Glyceraldehyde General flea. mulch as ghost-aldehyde has been proved to he an intermediate modest in the periods“ oxidation of moss, its reaction with periodete is of great importance. It is also a very simple sugar. In spite of its apparent silplieitm its reaction with periodato involves competition between fear simltaneoes reactions. Moreover theweeenceofglneraldehydediaerwpossimomieatethe situation. . 'nze reactions taking place in the oxidation of the mm an be repreeeated iv the relieving schematic representation: 87 (19) ' ° ) 110003 e nmo C801! 032011 2 O (neon , P mo 3 I m Ph# j) 2 Hoocn 03° 1: 3 . O ECHO the symbols P1, P2, P3 and PM ilioh will be used tbs-“31m the succeeding discussion to represent the various molar quantities of periodic acid used in the four reactions. In addition r1, r2, r; and rh any be allowed to represent the rates of these respective reactions. Ems r12r3 represents the ratio of the rates of reaction 1 to that of reaction 3. In reaction mixtures containing an excess or periodic acid, an increase. in concentration, produces a decreased rate in the first stage of the reaction and a semewhat increased rate in the last stage. me one general trend is observed in an oxidation with sodiun periodste with the exception that in this case the decreased rate with increased cmcentraticn holds to the end or the reaction and it is difficult to get the reaction to run to completion under the conditions or those ameriments. in increase in acidity of the solution also decreases the rate of the iirst stage oi‘ the reaction, but increases the rate of the last stage. In an acid solution the theoretical endpoint is quite 88 readily attained, but in near]: neutral solution such as is the case with sodim periodate the reaction is not comlete. mesa may all be pH effects if los pH's cause a decrease in initial rate. Hm, this would hard]: explain the decrease in rate due to increased concentration when using sodium periodste, inmoh :- this solution is and: sunny acid. more 1., of course, a decrease in p11 as the reaction proceeds and liberates formic acid. If a lee pa decreases the reaction then it appears difficult to under- standshythislaststam ofthe ”diaperiodate reactionshonld decrease more than that of corresponding reactions involving periodic acid, which is considerably more acidic. more is the probability memmmeawmunt reactions 2 andhshichneypossibly be catalysed by acid. Harem, reaction 1: represents the oxidation of moral, shich gave no evidence of an appreciable difference due to acidity when it see osidised W itself. In the reactions involving excess glyceralde-n hyde (Table XIV) there appears to be a very slight accmlation of ghoul when using sodim periodate, but this difference is so mall as to be almost negligible as compared with the greater differences in completion of the reaction as indicated in Tables III and 1111. Development of relationships when Meraldehyde is present in m. When considering the quantities involved in the reaction con- taining excess glyceraldehyde, an interesting series of relationships is found. Knowing the original molar quantities of glyceraldehyde and periodic acid and the production of formaldehyde and formic acid 89 by analysis and knowing by periodate titrnticns that the periodate reaction is canplets under these conditions, it has been found possible to calculate all the other quantities involved. m reference to equation (19) and by expressing all products and reactants in molar quantities the following relaticmhips are obviously truer. Glyoxnl produced - p3 - Pb ' Formaldehyde produced -.P3 + P2 Fomic acidprodnced-PleP242Ph Periodic acid con-mod (total) . ’1 4» p, + P3 o Pl; Glyceraldehyde remaining . Original glyceroldetwde - P1 - P3 Glyceraldelvde consumed II ’1 ‘5 P3 Ghoul produced - Periodic acid (total) - formic acid Glyceraldelvde consumed I- Formaldehyde e glycolaldehyde Glycolaldekwde produced a P1 - P2 From the preceding relationships the following equations m be developed: GlycolaJdehyde produced - 2 Periodic acid (total) - 2 3030 - 30001! Glycolaldelvde produced In 2 Glycol - 2 am 0 cocoa Glyceraldehyde consumed . Periodic acid (total) - Hon «- glroxal With the above relationships it is possible to calculate the molar quantities of all the products in the reaction. Since in all of these relationships there'are four unknowns (P1, .P2, P3 and P11) and since there are only three independent equations (All other relationships are dependent upon the three.), the problem has no direct mathematical solution based on the data available. However, if the ratio of 90 glyceraldehyde to periodic acid is increased prematurely to infinity a limiting ratio or r1 to r3 end“ to be reached since the relative acute of periodic acid represented Iv P2 and ’1; will tom to been less and P1 and P3 will represent practically the entire amount of periodic acid. Their ratio will then represent the ratio of the rates of reaction 1 to reaction 3. be calm headed "mum Radio. or r1313" in Table XIV represents the mallest possible ratio of ,1 and P3 {that is, the ratio 1'1ch Inst be equal to or greater than this value) indie calonlatedbymansotmeqaation, Ihichis dmlcped as follow: (a) Lot 13 - 1. (b) Then the ratio e! 1.1ch may be called a. (c) Then glyoereldehyde oormed (fit) «- glycolsldohyde, 1: r2 is neglected. cral‘ d (a) rzwhueoma-wr W "~- (e) fit a detenined from equation (c) should approach an erald asymptote as the ratio of c nod; increases. (1‘) Solving (c) for x: colald do 1 ' fiafie commed ~W It is evident from the calm headed "Minimum Ratio of r1313 that r1 is more rapid than r3 and that when the mpetition for periodete ions becomes sufficiently great the relative rates become more praninent as shown by the above ratio and qualitatively w a comparison betueen the relative quantities of fomic acid and romaldelvde or hy glycol- aldelwde and glycxal. 91 Fifect of a chm. & considering in Table XIV the dif- temnoeswhichare cmsedbyachangeinpfi, itisfmdthatm addition of perchlario acid produces mall increases in the ratio o! [mic acid to fmalddvde, of glycclaldehyde to glyoxal and in the minimum ratio or rltrJ. niece would all indicate a relatively moreased rate for reaction 1 or a relatively decreased rate for reactim 3. It would, of course, be possible for the added acid to increase or to decrease the rates or both reactions. The results ob- tdned with excess periodate (see Table XIII) which indicate a general rate decrease for the first stage of the reaction then the pH has been decreased 17 the addition of perchlorio acid, would support the idea at a general decrease in the rates of reaction 1 and 3. If this is the case, there nest be a relatively pester decrease of r3 than at 1'1. Hanover, inTableXIVachangeoprdoesnotsoentoafiect the glycolaldehyde produced. Therefore, 1! r3 is decreased more thm r1 a greater quantity of periodic acid cagit to have an opportunity of reacting as P1, thus producing a larger quantity of ghoclalderq-de. Melon, the quantity of glycolaldehyde produced does not seen to increase. moraine the rate of reaction of glycolaldehyde (r2) whincnasedbymincreaseinacidanddecreaoedWanincrease inpfi. Illisdeoreaseinr3andismreaeeinr2 cooldatleaat agree Iith the results obtained in Table XIII. This suggested increase in reacticn rate for glycolaldehyde in acid solution Malt profitably be checked, providing glycolaldehyde of nfl’ioient purity were available. As has been mentioned, a small 92 quantity of the impure compound was prepared from tartaric acid, but two subsequent attempts at preparation produced no yield. With glycoleldehyde available, a comparison of reaction rates in reaction mixtures of various laws should then throw sane ligzt on the problm. Buffered solutions have not been used in any of these emerinents, because of the effect of the buffers on the reaction. 00W of reactions run at various pH's maintained by buffers are practically valueless. Such cmsrisons m show only the effects of the buffers used rather than effects or pH. mfect of a d‘EE in concengation without m of Md” me to periodatg ratio. no effect of changes in concentration on the reaction of periodic acid and glyceraldehyde may be due to acoaupanying pH changes, that is, a more omentrated reaction udxtnre wold necessarily have a lower pH. h consideration of the variations in a reaction when the concentrations are decreased without chasm the ratio of glyceraldehyde to periodate (see Table XIV) , it sill be seen that dilution canoes a general increase in the ratio 'rlirB. This is also borne out by the increased ratio of formic acid to foetaldehyde and of glycolaldelwde to glyoxal. Both of them latter ratios would be increased by a relative increase in rh, which would increase the yield of woolaldelwde and decrease the yield of glyoxal. It met be remembered that am conclusions drama from Table XIV can give only a caparison of relative rates and can not give absolute rates. 93 The data in Table II using an excess of periodic acid indicste a taster general reaction at meter dilution, especially with respect to {mldeme production, which could be interpreted as a. more rapid reaction or glycolaldehyde (r2) or an increase in r3. However, mariner) with the dsta in table 117 shove that at meter dilutions there is case inorossc in moolsldehyde. mi- ie hardly cmpatible with a rare rapid reaction or glycoleldehyde (r2) unless eccmpmied hemeHhat meterimreaseinrl. in increase in r3 Without a corresponding increase in rh voila non an acouulstion of glyoxal. This is in hsrmom with the evidence. his reasoning now presents two alternatives, which could take place upon dilution, either one or both or which may take place: 1. in increase in r1 with a smewhst mailer increase in re. 2. An increase in r3 without a corresponding increase in ’h‘ be second of these alternatives does not seem quite as logical es the first inemoh as the ratio r1113 appears to increase with dilution. on first alternative does not explain the small increase in sliml. Therefore, the met logical explanation would be e combi- nation of the two. his conclusion is also in ayeement with the increased momldehyde conmmption at greater dilutions. Effect of increeaing Mereldghlde concentration. By increasing the Wehyde concentration while maintaining periodic acid con- centreticn, the pH is kept nearly constant. It will be observed then that the fmfldehyde and glyoxal yields are decreased and formic acid end glycolaldehde yields are increased many fold. 1111:! shows that r1 9h is considerably water than r3 and that an increase in glyceraldehyde concentration manly maplifies the opportunity for reactions 1 and 3 to take place rather than 2 or 1;. The fact that part of the glycer- aldohydo is in the form of a diner (In a 0.1 M solution at 0° 0 and at equilibrimn about 16% is in the form of the diner.) would tend to counteract this effect, but‘apparently even this is not great anon?! to provide opportunity for reactions 2 and h in solutions containing him ratios of glyceraldchyde to periodic acid. Effect of increasing agriodic acid concentration. its series in which the concentration of glyceraldehyde is kept constant and the concentration of periodic acid is Varied emphasises the same general trend as in the preceding series. However, it also would be affected by the change of pH as the concentration of periodic acid varies. herein-e, conclusions based on this series are not as clear, althougi the general trend emphasises the effect of the increasing glyceralde- hydsc periodic acid ratio rather than it does the effect or 133. One fact which has been proven without doubt in these series or reactions is the much poster rate of attack ts periodate on the cerl-carbiaol bond (01-03) than on the glycol bond (Cg-C3). file ratio of these two rates can not be definitely detemined, ht that it is greater than fire can be stated with a considerable degree of certainty. 95 SUMMARY 1. A spectrophotometrio method for determining the periodate ion in reactions involving the Malsprade reaction has been developed. the method is based on Wat of the optical densities at 222.5 nun- microns. Simple sugars, tonaldehyde and iconic acid do not Were. Bicarbonates do interfere. 2. The aechanisn or periodate oxidation of Q—glnoose has been elucidated. A small but noteworthy ”action of the glucose is oxidised in the open chain torn ad the mainder in the cyclic fora, producing an ester, 240ml glyceraldehyde, which then slowly hydrolyzes. This hydrolysis stage is the rate-determining step in the reaction. g-uabinoee has boon tend to follow a similar nechmin. 3. Periodate oxidation has been found to take place with 1,3-dio ketones, both cyclic and acyclic. The following canponnds have been found to react! acetyl acetone, l,l-dimethylcyolohexan-3,S-diome, 3-nethyl-2,h-pcntsdione and l,h.diphexvl-l,3-butsdione. The reaction produces carbon dioxide with unsubstituted 1,3-diketones, ht no formaldehyde or formic acid. 1 neon-him for the reaction has been postulated. h. A method of calculating glycolaldehyde and ghoul produced and glyceraldehyde consoled has been worked at for reactions involving 96 success ghosraldéxyde and in which formic acid and formaldehyde have been determined by analysis. This method has also been applied to a determination of the relative rates of attack on the bonds of glycer- aldehyde 0 S. 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