gm}... 53¢ 55” Sum- . .C} fi.‘ .82. .c '0'“ "t 0 Q an". 99:! ”our: 2 t g hit .0“. . IV...)— D- {L i! .. 1:." mm v 3 Q Q a: a- 3*. i 9.. 2f". .nm.” 1...... E. A? Mmffiu 3135 3:53:33??? . 3* ' a? . .._ . . EZOW‘E‘éEN“; EB Tfa g; '2 -; k, h 1 ‘AWQ ' $3 is f .a H 5‘“ ;‘.:§m€:£‘s i 1 ‘3‘ t? ‘13; C 335153 25‘ 2’ ‘33. 3%» Ci .3 h i . . .30 .8 ~ ‘q’. T. . . ‘1 y... t L 21'. r 5 .t I; I...“ E EWEC‘? {33? as A V3.3" V l L a ', ‘N y. «8 H:=2::__,:___:____:_ mmm m 92 ll; IIIIIIIIIIII IIIIIIIIIIIIIIIIIII ‘1 23 00678 6952 ABSTRACT THE EFFECT OF VARIOUS ADDITIVES ON COLOR DEGRADATION AND BROWNING OF INDIVIDUALLY QUICK FROZEN RED TART CHERRIES. by Shimon Mayak Red tart cherries (Prunus cerasus L. var. Montmorency) frozen as individually quick frozen (IQF) fruit underwent red color loss and oxidation or browning during 8 months of frozen storage. Pretreating the cherries by dipping into 500 or 1000 ppm solution of 802 for one minute before freezing decreased the amount of red color loss and browning with less color loss occuring at the higher concentration. Color loss and browning in cherries pretreated with 0.l% citric acid was similar to that obtained for the control while those treated with 0.l% ascorbic acid - 0.1% citric acid showed greater color loss and browning. Cherries frozen in 60% sugar sirup showed little or no color loss or browning during storage. Under accelerated oxidative defrosting conditions, red color loss and browning occured in all treatments. The rate and amount of color loss and browning was lowest in the cherries treated with 1000 ppm SO and greatest in the ascor- 2 bio-citric treated fruit. Color loss and browning occurred at an accelerated rate in the sirup packed fruit and after 60 minutes was greater than that of the control samples. Individually quick frozen cherries packed in 30 lbs. friction top cans retained more red color and showed less browning than those packed in 1 lb. polyethylene bags. Shimon Kayak No 802 could be detected in 802 treated fruit either by the Monier-Williams distillation method or by treatment with cold NaOH prior to distillation. Thirty-two to forty-four ppm of 80 were retained by 2 either the control or 802 treated fruit held in 802 solutions containing 80, 160 and 300 ppm for 16 hours at 32°F before distillation. The addition of 25 ppm 802 prior to defrosting under highly oxidative conditions prevented red color degradation in the control samples but did not completely prevent oxida- tion of phenolic compounds or browning. Twelve ppm 802 added prior to defrosting prevented color destruction and browning in the cherries pretreated with 1000 ppm 80 while 25 ppm added 2 Just before thawing resulted in the bleaching of fruit. THE EFFECT OF VARIOUS ADDITIVES ON COLOR DEGRADATION AND BROWNING OF INDIVIDUALLY QUICK FROZEN RED TART CHERRIES BY Shimon Mayak A THESIS Submitted to Michigan State University in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Department of Food Science 1965 ACKNOWLEDGMENTS The author wishes to express his sincere appreciation to Dr. Clifford L. Bedford for his valuable guidance and helpful suggestions throughout the course of this research and during the preparation of this manuscript. He is deeply indebted to Dr. Perciles Markakis for his helpful discussion and criticism, and to Dr. Paul K. Kindel for his critical evaluation of this manuscript. The author is deeply grateful to his wife, Batia for her continuous understanding and encouragement throughout my graduate program. 11 TABLE ACKNOHLEDGTENTS . . . . LIST OF TABLES . . . . IST 0? FISJRES . . . . INTRODUCTION. . . . . . LITERATURE REVIEV . . . METHODS AND NATERIALS . RESULTS AND DISCUSSION SUMMARY . . . . . . . . LITERATURE CITED. . . . APPENDIX. . . . . . . . OF CONTENTS ill Table Stability of red color values in alcoholic solution . . . . . . . . . . . . . . . . . . . Stability of red color values at pH 3.4 and pH 5.0. . . . . . . . . . . . . . . . . . . . Color loss under accelerated oxidative defrost- ing conditions for the various treatments and storage time . . . . . . . . . . . . . . . . . Effect of various treatments on the oxidation of phenolic compounds during frozen storage, and subsequent accelerated oxidative defrost- ing conditions. . . . . . . . . . . . . . . . Comparative discoloration rates in red color absorbancy at pH 3.4 and as difference between pH 3.4 and pH 5.0. . . . . . . . . . . . . . . Percent color 1088 based on absorbancy values determined at pH 3.h and by difference between pH 3.0 and pH 5.0. . . . . . . . . . . . . . . Effect of various treatments and different packages on the oxidation of phenolic compounds. . . . . . . . . . . . . . . . . . . Milligram SO recovered from frozen cherries 2 stored with known amount of 302 . . . . . . . iv ILL 21 DJ \ LIST OF TABLES (continued) Table 9 10 Page Effect of added 802 on color loss under accelerated oxidative defrosting conditions . . 35 Effect of added 302 on phenolic compound oxida- tion under accelerated oxidative defrosting conditions. . . . . . . . . . . . . . . . . .1. 35 LIST OF FIGURES Figure 1 Absorption curves of red color values during storage, pH 3.4. . . . . . . . . . . . . . . . Comparative discoloration rates of red color, absorbancy at pH 3.4 and as difference between pH 3.4 and pH 5.0. . . . . . . . . . . . . . . Effect of various treatment on the rate of red color destruction during accelerated oxidative defrosting conditions. . . . . . . . . . . . . Effect of container on red color degradation in control cherries, under accelerated oxidation conditions. (Absorbancy at pH 3.4 and as difference between pH 3.4 and pH 5.0.) . . . . vi 18 26 INTRODUCTION The red color of red tart cherries (Prunus cerasusJ L. var. Montmorency) constitutes an important quality factor ‘readily evaluated by the consumer, and therefore has a signi- ficant effect upon the value of the product. The loss of red color and subsequent oxidation or brown- ing of the fruit tissue is related to the extent of bruising during harvest and the temperature at which the fruit is held. Lowering the temperature decreases the rate of color loss, but even in frozen storage color loss will continue 1 throughout the storage period. The loss of red color can be minimized by decreasing the exposzre of the fruit to air. In the past few years there has been an interest in freezing red tart cherries as individually quick frozen fruit similar to the well established process for vegetables such as peas, lima beans, snap beans and cut corn. It is felt that this system of freezing would give a product that would have the advantages of being free flowing, reduce shipping and storage costs and permit multiple uses of the fruit by the consumer. There is no date available on the effect of freezing and frozen storage on the quality of individually quick frozen cherries. {his study was undertaken to determine the effect of pretreafing the pitted fruit with sulfur dioxide, ascorbic acid and citric acid on retention of red color and quality of 1 2 individually quick frozen cherries. LITERATURE REVIEW Roger (1940) reported that individually frozen cherries do not retain their original bright red color as well as cherries packed and frozen in 30-1bs. cans. Lee (1949) found that, when the fruit was packed in small containers and covered with 50-60% sugar sirup, the rate of freezing had little or no effect on the quality of the fruit after six months storage at 00F. Guadagni (1963) reported that rapid freezing of 30-lbs. cans of cherries (5+1), reduced the browning of the surface layers and the transfer of the red color from the skins to 'the expressed surrounding Juice. ANTHOCYANINS The anthocyanin pigments of the red tart cherries have been identified by Li and Wagenknecht (1956) as cyanidin- 3-rhamnoglucoside (antirrihinin); and cyanidin-3-diglucoside -(mecocyanin). These pigments are phenolic compounds and are readily oxidized or reduced with the loss of red color. Sondheimer and Kertesz (1952) reported the oxidation of antho- cyanin by hydrogen peroxide to a colorless product in straw- berry Juice when H202 was added. Although the formation of hydrogen peroxide has not been demonstrated, they suggested that it was formed during the oxidation of ascorbic acid catalyzed by Cu or Fe. u Meschter (1953) reported that Fe and Cu increase ascorbic acid oxidation and that the oxidation products accelerate anthocyanin destruction. A rather different mechanism was reported by Huang (1955), in which crude enzyme extracts from ASpergilli hydrolyzed blackberry anthocyanins to anthocyanidins and sugar followed by spontaneous transformation of the aglycone to a colorless derivatives. Wagenknecht et a1. (1960) isolated crude anthocyanase from red tart cherries. It is believed to participate in the early stages of scald (red color loss) in red sour cherries through destruction of anthocyanin pigments. The enzyme required the presence of oxygen for its activity and was acti- vated by the presence of catechol. Scheiner (1960) reported that the anthocyanin decoloriz- ing activity of cherry crude homogenates is almost doubled by the addition of catechol and that purified enzyme prepara- tions are almost completely inactive unless catechol or some other o-dihydroxyphenol compound was present in the reaction mixture. Catechol oxidase activity and anthocyanin decoloriz- ing activity were found in all the fractions tested. He sug- gested that the anthocyanins were non-enzymatically oxidized by the quinones produced in the enzymatic oxidation of dihydroxy- phenol compounds. Peng and Harkakis (1963) using mushroom tyrosinase found that in the absence of catechol the reaction was very slow, but in the presence of catechol the rate of mecocyanin decolori- ation increased rapidly. Grommeok and Markakis (1964) showed that horseredish peroxidase oxidized cherry anthocyanins to a colorless form, and the maximum rate occured in the presence of H202 at -4 to 10"3 M. concentration of 10 Although phenolases and peroxidases were not purified from cherries, their activity has been reported in cherries, (Bedford, 1963). PHENOLIC COMPOUNDS The.mechanism of the oxidation of phenolic and poly- phenolic compounds and subsequent polymerization with the formation of brown pigments has been summerized by Mason (1959). Briefly, when catechol is oxidized in the presence of poly- phenoloxidase, the first step in this oxidation produces o-benzoquinone, which may polymerize to form simple melanins. Guadagni (1949), using the method described by Rosenblatt and Peluso (1941) for total phenolic compounds on a water extract of frozen peaches, pointed out that it is reasonable to assume that the loss in phenolic compounds as measured herein is a fairly good estimate of the degree of enzymatic browning. He found that 70-80% of the total phenol compounds present in a water extract were converted to dark brown sub- stances._ Hillis and Swain (1959) extracted phenolic compounds from Victoria plum tree with absolute methanol and 50% methanol, and found that the absolute methanol fraction contains simple 6 phenols like chlorogenic acid, and also mono or oligomeric leuco-anthocyanins. The aqueous mathanol contained higher polymeric forms. However, the higher polymeric forms were not necessarily insoluble in absolute methanol, but bound via hydrogen bonds to cell walls or protein in the plant material, and were only released when partial rehydration with an aqu- eous solvent mixture broke these bonds (Goldstein and Swain, 1953). It might suggest that extracting with water, although less selective, would provide a simple extraction procedure for studying the change in phenolic compounds due to enzyma- tic browning reaction. SULFUR DIOXIDE Sulfur dioxide and the salts of sulfurous acid have been found to be very effective in the retardation of enzymatic and non-enzymatic browning as well as preventing undesirable color and flavor changes in fruits (Joslyn, 1954) and(Ponting, 1960). 802 is also used to bring about the irreversible bleaching of cherries (Marashino) (Sullis, 1931) or the reversible bleaching of anthocyanin pigments (Jurd, 1964). The chemistry of 302, sulfurous acid and its salts have been reviewed by Joslyn and Braverman, (1954) . Although it has been used for a long time, the mechanism involved in retardation of discoloration was not well established. Hamburger and Joslyn (1941) reported that 502 prevented darkening by being readily oxidized. Joslyn and Hohl (1948) suggested that it act as an antioxidant, utilizing the oxygen 7 available. Johnson and Johnson (1952) reported that it decreased the enzyme activity and this was supported by Ponting (1960) who reported that polyphenolase activity was completely inhibited in the presence of 10 ppm 802 in buffered solution containing catechol. However, if the enzymatic oxi- dation was allowed to proceed until some quinone is formed, then the addition of 5 ppm 802 will not alter the enzyme acti- vity because the 802 reacts with the quinone. Demair et a1. (1960) reported that enzyme inactivation is reversible and is not related to 802 oxidation. Sastry et a1. (1961) found com- plete peroxidase inhibition in the presence of 25 mg% 302 at pH 4.0. Goodman and Markakis (1963) reported that if the enzyme and 802 were preincubated, lower levels of 302 had to be added to the Juice in order to obtain inhibition similar to that of no preincubation- Embs and Markakis (1965) reported that the immediate 802 inhibition of browning is due to the formation of a quinone 802 addition compounds and preventing polymerization of malanin pigment. No oxidation of 802 was found. 502 DETERMINATION Sulfites in fruit and vegetable tissue will exist as solution of sulfurous acid in water, as the free bisulfite ion, as the free sulfite ion, and as more or less tightly bound $02 in the form of the hydroxysulfonate (Joslyn, 1954). Ponting (1960) pointed.out the possibility that only free 302 8 inhibits the enzyme, although almost always the total 802 is measured in fruit (the Monier-Williams distillation method). It has been consistently observed that the quantity of 802 in the fruit will decrease during storage (Burton et a1., 1963). Ponting (1945) reported that the fruit texture and low temperature, employed in freezing fruit prevents penetration of 502 much below the surface, thus large portions of the enzyme remain potentially active, and upon thawing, the enzyme willoxidize the 802. There is a possibility of sulfur dioxide loss other than by oxidation to sulfate. Stadman (1948) reported loss of 80 from dried apricots in absence of 2 oxygen. Burton et a1. (1963) concluded, that p -sulphonalde- hyde formed in the reaction of 802 and unsaturated aldehydes, ,,o NaH303 I; OH R-CHzCH-C —————> R-CH=CH-c’ T’ ‘\H Efast \SOBNa haHDOB (slow) I I ,0 ) /OH R-CH-CHZ-C\ H 633.0 enm,6 mmn.o 0mm.o mum.o mno.a maw.o mm. M ansqo mem.: amn.6 336.6 6s6.6 mso.a 666.6 6a- . ama.6 mm6.6 .mm.6 mn6.6 mam.6 ~66.H «H6.6 6s, mme.6 asm.e Lem.6 656.6 66n.6 66H.H NH6.6 awn mne.6 oom.6 mun.6 666.6 Hmm.6 N66.H NH6.6 66m A 666.6 666.6 .nm.6 a66.6 Hmm.6 666.a 666.6 was a .6.“ m6. .6.m m6. .6.m ad. .6.m m6. .6.“ ed. .6.m m6. .6.m.md. .6.m m6. meanness sonoAm M\ma mam< om nu ow m: on ma Adds. 6666a nowaxlnspa. mafia .o.m ma cam :.m mm as mofiadb uoaoo con «0 hpaaandpm .N 6.666 15 Comparisons were made on representative samples of all treat- ments at the end of 8 months storage period between cherries packed in 30 lbs. friction t0p cans and 1 1b. polyethylene bags. Absorbancy values were determined at one pH (3.4) and as a difference between two pH's (pH 3.4 - pH 5.0), according to the procedure recommended by Sondheimer and Kertesz (1948). The rates of anthocyanin degradation and oxidation of phenolic compounds under accelerated oxidative defrosting conditions were determined using the procedure described by Guadagni et al. (1949), with the exception that the tempera- ture was not raised to 70°F. Representative samples were also blended with 12 and 25 ppm SO to determined the effect of 2 added 802 on the rate of oxidation. Total Phenolic Compounds The total phenolic compounds were determined on the alco- holic extract using the method described by Rosenblatt and Peluso (1941). Transmittancy was measured at 660 mu using the Evelyn colorimeter. The absorbancy curve of the blue color produced, using the Bausch and Lomb 505 Spectrophoto- meter showed no peak in the range of 640 mu to 825 mu. Sulfur Dioxide Determination 302 content was determined by the Monier-Williams dis- tillation method (A.0.A.C. 1960). 50 gm of frozen cherries were transferred to the distillation flask, followed by 300 ml. distilled water and 20 ml. concentrated HCl and distilled into 16 3% H202 long enough to allow 30 minutes boiling time. The possible presence of "bound" sulfur dioxide was determined by incubating 50 gm of cherries with 100 ml (0.25 N) NaOH for 30 minutes, acidifying and distilling for 60 minutes. (Dis- tillation times up to 120 minutes did not increase the yield of 802 in either procedures.) Representative 50 gm samples of the control and cherries pretreated with 1000 ppm were held in 100 ml of 80 solution 2 of different concentrations for 16 hours at 32°F. Then the 302 content was determined by distillation to obtained data on the amount of 502 that could be recovered. Blank determin- ation were made on each of the 802 solutions. RESULTS AND DISCUSSION Color changes during frozen storage The most readily observable visible changes in pitted red cherries frozen without any pretreatment or packing medium were the loss of red color and browning of the flesh. Color loss occurs throughout the storage period in the treatments (figure 1) and (data appendix table 3) The cherries pretreated with KWO ppm 802 showed the least red color loss, followed by those treated with 500 ppm 302. The citric acid treatment was similar to the control, ascorbic acid - citric acid treated samples had greater color loss than those of the control. Sondheimer and Kertesz (1952) suggested the formation of H202 during ascorbic acid oxidation would rapidly oxidize anthocyanin pigments to a colorless product. Since the cherries were not protected from exposure to oxygen, rapid oxidation of ascrobic acid might be expected. Unfortunately during the storage period a breakdown of the freezer occured, the freezer temperature reached 400F before repairs were made and the temperature again reduced to ~50F. Although such storage conditions are unusual, it has been pointed out by Hunter (1953) that the temperature of frozen fruit during tranSportation, storage and distribu- tion might reach as high as 40°F. During the period the cherries were defrosted and refro- 1? 18 newsman an» no szomxmcan on 056 wcapmoahmn t. A6666 6.. nzHa mm .666 6666 6. 66m 6am 66a 66H 6NH 66 66 .66 6 l _ _ 4 _ J 3 g? a . .I +. I/ II. D + flu 1 4 s . / a, +: 1%., 6// ao—o’ 4 :: .22 . o /.. . ../ A_. a.. ’.+ QSHHm ’MQ HmwSm C / , macs oaupao I macs canaocm<.6 cl .4 Houpcool... +1 O macs casuaoID 0: N66 :66 666 .< 4.. mom Edd 83 ad on m.o (\I o H .:.m ma .mmmAOpm msaHSp modam> Hoaoo can go m0>h50 moduahomnm .H chamam w.H setxreuo uszox; mB/gISV 1 \1 ) zen, the red color loss in the 802 treated fruit was much less than in the control or other treatments. Peng and Markakis (1963) reported that the breakdown of the anthocyanin pigments was mainly by reaction with quinones, the oxidation products of phenolic compounds. The increase in enzymatic activity during the rise of temperature would result in increased formation of quinones, which would be available to react with the anthocyanins. Embs and Markakis (1965) indica- ted that 802 reacts with quinones to form addition compounds. This would reduce the amount of quinones free to react with the anthocyanin pigments in the 802 treated fruit, and there- fore reduce the amount of red color destruction. The relative rates of red color loss during accelerated oxidative defrost- ing conditions after various periods of storage are given in table 3. The relative rates depend upon and vary with the red color levels (substrate concentration). The control and the ascorbic - citric acid pretreated cherries showed the highest color loss rate during accelerated defrosting conditions after 30 days of frozen storage with about 38 and 46% loss of red color after 30 minutes of blend- ing. (table 3). The lowest red color loss rate occured in the fruit pretreated witthOO ppm 80 The relative red color 2. loss rates decreased in the control, ascorbic - citric and citric acid treated fruit during storage. The rates increased slightly in the SO treated cherries with blending time which 2 is in agreement with the results reported by Goodman and 20 Harkakis (1965), who observed increasing rates of phenolic compounds oxidation with time at low concentrations of 802. Quinone formation during storage in excess to those bound with SO2 will result in increasing discoloration rates. .muHUCmHQ maaplo um Hoaoo Hmapaca on pmpdamh mm popmHSOHmo wQHUCmHD mcauSU mmoa .Loom.av msdnmchm Houmm mcaasmno Umppaa no 65Hm> Hoaoo HQWpHC .moahhmso £06069 86H km on Umpmaca mm 66pmaso mo mmoa m 6666650660 as mam as g 66.66 66.66 6666I, 66»6. .w6umw. 66666 6 66.66 666.6 66.66 666.6 66.66 666.6 66.66 666.6 66.66 666.6 66 66.66 666.6 66.66 666.6 66.66 666.6 66.66 666.6 66.66 666.6 6H 66.66 666.6 66.66 666.6 66.66 666.6 66.66 666.H 66.66 666.6 6 666 III w: 6.. HEDHP 66.66. .66.6H pbww. 6:6666 6666 a 66.66 666.6 66.66 666.6 66.66 666.6 66 66.66 666.6 66.66 666.6 66.66 666.6 66 66.66 666.6 66.66 666.6 66.66 666.6 6 666 msabmep 66.6 66.66 66.6 66.66 66.6 666666 6666 a 66.66 666.6 66.66 666.6 66.66 666.6 66.66 666.6 66.66 666.6 66 1 .6 66.66 666.6 66.66 666.6 66.66 666.6 66.66 666.6 66.66 666.6 66 66.66 666.6 66.66 666.6 66.66 666.6 66.66 666.6 66.66 666.6 6 66 ESEBTCEHSS 66.6w 6mM66 66.6 66661 66.66 6666 a ... 66.66 666.6 66.66 666.6 66.6 666.6 66.66 666.6 66.66 666.6 66 66.66 666.6 66.66 666.6 66.6 666.6 66.66 666.6 66.66 666.6 66 66.66 666.6 66 6.6.H 66.6 666.6 66.6 666.- - 666.6 6 66 - A.GHS. 6666 a 6666 6666 a 6666 6666 a 6M6< 6666 m 66H6< ..6666 a #6666 me.» .6666. 6666 666666 6666 oawpaouoannoowa and 6666 66 and 666 66 6666266 meaeeoam mafia mummapcmap msoaam> on» How wQOapapsoo wcmumcaumn mbauwpdwo dcpmncamooc 666w: mmoa aoaou .osap mwdaoum 6cm .6 66666 n r. , ’1' I. 5.- The oxidation of the phenolic compounds was not determined for all storage periods, but the results obtained for two of them 34 and 240 days, (table 4) showed the same trend as red color destruction. Table 4: Effect of various treatments on the oxidation of phenolic compounds during frozen storage, and subse- quent accelerated oxidative defrosting conditions. Storage time (dayS) 34 240 Blending time (min.) 0 15 30 0 15 30 Treatments A660mu/gm frozen cherries Control 4.416 4.256 3.880 3.660 3.348 3.312 so2 500 ppm 4.672 4.576 4.224 4.032 3.720 3.672 302 1000 ppm 5.326 4.960 4.956 4.260 4.080 3.960 ' Ascorbic-citric 4.366 3.952 3.520 3.648 3.432 3.024 ‘ Citric 3.904 3.632 3.312 3.216 3.036 3.060 The values are averages of 4 replicates Sondheimer and Kertesz (1948) stated that measuring absorbancy at one pH as a measure of red pigment content is not very accurate, particularly at an advance degree of oxidation, because of the increasing proportion of brown pigments which contribute to the absorbancy. He suggested that the difference between the absorbancy values measured at two pH levels is truer representation of anthocyanins content. 23 Figure 2 shows the relationships between absorbancy values obtained at pH 3.4 and those obtained as a difference between pH 3.4 and pH 5.0. Although the absorbancy difference value indicated higher % loss of red pigment as compared to absor- bancy measured at one pH level, the rates of destruction do not differ appreciably. These results indicated that absorbancy determination at one pH were suitable for measuring rate of red color destruc- tion. The percent loss of red color and the percent loss dur- ing blending calculated on the basis of the two procedures are given in table 5, for the control and HKD ppm 302 treated fruit. The red color losses based on the absorbancy values measured at pH 3.4 were 72 and 46 percent reSpectively for the control and 1000 ppm SO pretreated fruit after 150 minutes of blend- 2 ing and those based on the pH difference measurement were 87 and 56 percent. The rate of red color loss in the various treatments after 240 days of frozen storage under accelerated oxidative defrost- ing conditions is shown in figure 3 (data appendix table 4). The cherries pretreated with 1000 ppm 302 showed the least red color loss follow by 500 ppm 802, citric acid and control, sirup and ascorbic acid-citric acid. The sugar sirup packed cherries while retaining the highest red color throughout storage period, under highly oxi- dative conditions showed very rapid red color destruction. 24 Aoon.av wsammmuu Honda msam> depHCa on vmpwama mm avocadoamo .wmoa & 5.:HEV mafia msfichHm omH ONH om cm on ma 0 « _ _ _ _ _ HOHDCOU ill||\\\LQ\\\\\\\\\ <\I\I\\b Ao.n ma .. :8 m3 .o.n mg was s.m ma noozpmn cocohmmuav mm cam d.m mg on mocwnaomnw .Hoaoo can no moody scandaoaoomac obapmawasoo “N opswam o: co om OOH g/ 8801 .0 C/ 0.... 0.0- .mmpmodaaoh 0 mo mommampm can mosam> .0005.0v 00000000 00000 0:00» 0000000 on 0000000 00 0000000000 .0000 000.00 0:00:002 mo mE0puo pm mSpr Hm0u0c0 on 0000000 00 0090050000 .wsaquHn wQHHSU 0000 0 00* R *# .moHHaoso £00009 no swam you as man 90 mp025 humanaomn< * 00.00 00.00 000.0 00.00 00.00 000.0 000 00.00 05.00 000.0 50.00 55.00 000.0 00.00 00.50 000.0 50.00 00.05 050.0 000 00.00 00.00 500.0 00.00 00.00 000.0 00.00 00.00 000.0 05.00 00.50 000.0 000 00.00 50.00 050.0 50.00 50.00 000.0 50.00 00.00 000.0 05.00 00.00 000.0 00 05.00 00.00 000.0 00.00 00.50 050.0 05.00 05.05 000.0 00.00 00.00 005.0 00 00.0 05.00 000.0 00.0 00.00 000.0 00.00 00.05 050.0 00.00 00.00 000.0 00 00.0 00.00 000.0 00.0 00.00 050.0 00.00 00.00 000.0 50.0 . .00 000.0 00 00.0 00.00 000.0 . 00.00 000.0 00.0 55.00 050.0 00.0 .00 000.0 0 mCH mC0 w:« a: A.:08v m0cm0n -00000 muco0n -00000 0500 :0050 u00nsr c0050 wC0070 mn0 000 & mmoa mHm¢ mmoa w mmOH R mHm¢ mmoa m mmoa m mHm< mmoa w mmOH m nan< lunoam tit #* t 00.0 00-0.0 000 00.0 000 00.0 00 u 0.0 000 00.0 000 002000900Q mononmmmao E00 0000 000 0000000 .o.m mg and 0.m mm newspon cocoamupdd mm was 0.m mm as mocwnpowpm Hoaoo Umh :0 momma scandaoaoom00 o>0uwnmmaoo «m candy 26 A.C0Ev 050p 0:00Cm0m 00 00 00 0000 000000 I 0000 00nhoom<|o _ 0 _ O 0.00.000 00.00%... Hopunool+ 00cm 00np0onD 000 000 000 14 +0 NOn 0.85 000.0100 ( .0200900200 weaumonwov o>0umdwxo nopMHmHooom wc0H:0 :00posapmou .HOHOO UQH .HO QDGH OED CO Ugmfipwmhp mSOde> .HO povhhfl "m 0H5000 0...” N30 W'°€ Hd 6mg? gtgv 27 The percent loss of red color for the various treatments calculated on the basis of the two procedures is given in table 6. The difference in percent red color loss between the two methods for the control, SO2 and citric acid pretreated cherries are relatively constant for each of the blending periods and are about 17, 10, 3 and 11% higher reSpectively for the losses determined by the difference method. The dif- ferences in the losses determined by the methods increased With blending time for the ascorbic acid - citric acid pre- treated fruit which might be due to increased formation of brown pigments as pointed out by Hodge (1953). No explana- tion could be given to the increasing difference between the two red color measuring methods as time of blending of the sirup packed fruit increased. The percent loss of anthocyanins after 2&0 days of frozen storage was 6, 32, 50, 52, 61 and 62 respectively for the sirup packed, 1000 ppm 802, 500 ppm 802, citric acid, ascorbic acid - citric and control fruit. After 60 minutes of blending the losses were 86, #3, 58, 67, and 98 percent reSpectively. _§§ect of containers The cherries stored in the 30 lbs. friction tOp cans showed less red color loss and browning than those stored in 1 lb. polyethylene bags for all treatments (figure a), and (data appendix table 4). In addition the rate of red color loss under accelerated conditions of oxidation was lower for .mmpmoaaamh 3 no mowmhmbm ohm modam> use .Ammm.ov madmoonm 909mm msam> Hmapdsa op dopmaon mm UmpdaSOHmo .mmoa & ** .Aoom.av madumohm Hmpum ozam> Hmapdsa ou Umpmach mm dopmadoamo .mmOH R * mm.m 0H.0m mm.mm oa.m0 mm.0: nm.:: mm.om NN.0 aspam n0msm ace 00.sm 30.0e mm.0m 0a.mm mo.m0 0m.mm am.H0 00.m: Ama.o u ma.ov.0dom oaapdo I cannoom< mm.e0 mm.:m m0.mm mm.m¢ Hm.mn oe.m: 3H.mm mm.aa Ra.o enom 0anao MW em.m: 00.nm 00.0m mm.mm sm.:m NH.Hm Hm.mm H:.mm sac oooa mom 0m.mm s:.u3 om.am em.ms 3H.mm H:.N: 00.0m H:.o: sag con mom Hu.me as.mm 00.05 mm.mm m2.n0 mm.on om.H0 mm.ms Houpcoo mmoa R mmOH R moOH w mmoa R mmoH m mmoH m mmoa m mmoa & uncapmoha at t ** t “0.0 mg Ao.m ma Ao.m ma Ao.m ma -s.m may As.m mac -:.m may A:.m may -:.m mac A¢.m may -:.m may A3.m may commaommHo cosmhmmmdo mosmaommdo mosmaomuam A.:Hav 00 on ma 0 05H» weaesoam .o.m ma cam :.m ma soozpon mononommdo an use :.m mg no omsaspmpmc mmsHm> mocoQHOmnm no woman mmoa Hoaoo pcmonom «0 0Hn09 ASIS/gm frozen cherries 29 Figure u: Effect of container on red color degradation in control cherries, under accelerated oxidation conditions. (Absorbancy at pH 3.4 and as difference between pH 3.4 and pH 5.0) 1.0 +— o on I p? 3.4 006'— Oeuv— . .\ \ >pH 5.0) C) 0.0 I 1 l l 0 15 30 60 Blending Time (min.) 30 the fruit stored in the 30 lbs cans. These results are in agreement with those of Guadagni gt al. (1957), who reported much less red color loss of red tart cherries in hermetically sealed containers as compared to metal and paperboard con- tainers. Although the friction top cans were not hermetically sealed, the amount of oxygen available would be less per unit of fruit than in the polyethylene freezer bags. It was also found that the bags were not completely vapor proof and there- fore would permit a continual supply of air to the product due to breathing action during storage. This is also amply demonstrated by the fact that fruit packed in sirup in herma- tically sealed containers showed very little loss of red color. The increased rate of the red color destruction in the fruit packed in bags could be due to a greater accumulation of quinonees during frozen storage. Scheiner (1960) and Peng and Markakis (1963) have prOposed that the main destruction of anthocyanin is by quinones and that the amount of quinone accumulation is dependent upon oxygen availability. The oxidation of the phenolic compounds in the various treatments showed higher degree of oxidation (lower values) in the'cherries packed in 1 1b. polyethylene bags as compared to those packed in 30 lbs. friction t0p cans. (table 7). .momeaHmon : no mowmumbm ohm modamb one moahpmso nmuonm Ew\ mmmp onHmSpomHog .Da H omo.0 «00.: 00m.n emxomc aspam 300.0 mm0.m 000.0 0000 ownpao soa9900m< 000.m 0mo.m 0am.m enom cappao 000.0 000.0 000.: add 0000 mom 000.0 000.0 000.0 000 000.000 mam.m mem.m 000.m Hoppeoo mucoapmmhe A.:Hav on ma 0 0aapfwcdee00m mado mpH om .wUQdOQSoo oHHosmsa mo sodomoaxo 0:» no mmwmxoma pcopmpmac cam mpCoEpmmhp msoanmb no poowmm an canoe J O Sulfur dioxide Determination Since 802 proved to be effective in decreasing the loss of red color and browning of the fruit, it seemed desireable to determine the residual amount present in the fruit. No objectionable flavor could be detected by sensory evaluation and there was no visual bleaching of the anthocyanins. No 802 could be recovered from any of the 802 treated fruit either by direct acid hydrolysis (Monier-Williams dis- tillation method), or by the combination of saponification with sodium hydroxide and acid hydrolysis. Distillation of known concentration of SO solutions gave recoveries of 98%. 2 These results indicated that either the 802 had been oxidized as prOposed by Ponting (1960), or it had combined in such a way that it was not released during distillation. In eXperiments to study the unrecoverable 502 cherries of both control samples and those treated with 1000 ppm 802 prior to freezing were stored in 802 solutions for 16 hours at 32°F. The results obtained showed that from 1.6 to 2.2 mg of 502 (30 to #4 ppm) retained by 50 gm of frozen cherries and not released during distillation (table 8). 33 Table 8: Milligram SO recovered from frozen cherries stored with known amount of 302. Control mg 502 added mg 802 recovered Difference 8.00 (0.00)1 6.08 (0.32) 1.92 15.68 (0.60) 13.40 (1.28) 2.24 30.72 (1.28) 29.12 (1.60) 1.60 802 pretreated cherries 7.68 (0.00) 6.08 (0.32) 1.60 16.32 (0.00) 14.40 (0.32) 1.92 1The range of concentrations. The values are averages of 5 replicates. Burton and McWeeny (1963) suggested that 802 reacts with unsaturated aldehydes forming p-sulfonaldehydes, which will not release SO2 upon acid hydrolysis. This appears to be a likely mechanism leading to a reduction in the amount of measurable sulfite in stored food stuff. Effect of added SOQ on color loss and oxidation under accelera- ted conditions. Since the cherries pretreated with 802 prior to freezing lost some of their red coloz and browned under accelerated conditions, it was of interest to determine the amount of 802 required to prevent discoloration during blending. _ Control as well as fruit pretreated with 1000 ppm 302 were used. SufficientCL3% NaH803 solution was added to obtain 12 and 25 ppm of 802 in the mixture. Twenty five ppm 802 31+ prevented red color destruction during 60 minutes of blend- ing in the control samples, but did not prevent the oxida- tion of the phenolic compounds, although the rate was lower than without addition of 302. Higher total phenolic compounds values were obtained upon addition of 302 (tables 9 and 10). No explanation can be given for the increased phenolic compound value. The absorbancy values obtained using a pure 25 ppm 80 solution (blank), indicate that neither the increase 2 nor the reduction in phenolic compounds value during blend- ing could be accounted for by the reaction between the Folin- Denis reagent and 802. One might speculate, the addition compounds of quinones and 302, hydroxysulfonates, might react with the Folin-Denis reagent. However this is questionable since similar addition of 802 to the cherries pretreated with 1000 ppm 302 did not give an increase in phenolic compounds value. Twelve ppm 802 added during blending almost completely prevented red color destruction and phenolic compounds oxida- tion in the 1000 ppm 802 pretreated cherries; 25 ppm 802 bleached the cherries. Both 12 and 25 ppm 502 prevented the oxidation of the phenolic compounds during blending. These results indicate that it might be more desireable to use slightly higher concentrations of 802 in the pretreatment of the cherries prior to freezing. 35 Table 9: Effect of added 502 on color loss under accelerated oxidative defrosting conditions. Sample Control 802 1000 ppm 302 added in (ppm) 0 25 0 12 25 Blending time (min.) A515 mu/gn frozen cherries 1 0 1.128 1.123 1.411 1.382 1.166 15 1.070 1.114 1.334 1.382 1.152 30 0.974 1.128 1.248 1.363 1.195 60 0.725 1.114 1.138 1.320 1.186 The values are averages of 4 replicates Table 10: Effect of added SO on phenolic compound oxida- tion under accelergted oxidative defrosting con- ditions. Sample Control 802 1000 ppm H20 802 added in (ppm) 0 25 0 12 25 25 Blending time (min.) A660 mu/gm frozen cherries 0 3.6961 4.104 3.816 3.744 3.780 0.079 (0.053)2 15 3.480' 4.008 3.684 3.756 3.768 0.053 (0.053) 30 3.048 3.864 3.504 3.684 3.816 0.000 60 2.338 3.492 3.252 3.660 3.756 0.000 2The range of values The values are averages of 4 replicates SUMMARY 1 Tne effect of various chemical additives as to their efficiency in preventing red color degradation and phenolic compounds oxidation in red tart cherries frozen by indivi- dually quick freezing procedure was studied. Color loss and phenolic compounds oxidation occurred throughout the 8 months storage period in all the treatments. The cherries pretreated with 1000 ppm 302 showed the least color loss, followed by the 500 ppm SO pretreated cherries. 2 The citric acid treatment gave results similar to the control, ascorbic acid - citric acid pretreated cherries had greater red color loss than those of control. Cherries frozen in 600 sugar sirup showed little or no red color loss during frozen storage. The oxidation of phenolic compounds followed the same trend. Under accelerated oxidative defrosting conditions red color loss and oxidation or browning occurred in cherries in all treatments and the rate of change followed the same order as found for the treatments during frozen storage. Cherries frozen in 606 sugar sirup showed a very rapid rate of red color destruction and oxidation of phenolic compounds with increasing blending time. The various pretreated cherries packed in 30 lbs. fric- tion top cans retained much better red color than those packed in 1 1b. polyethylene bags. In addition the rate of antho- cyanin pigments destruction under accelerated oxidative 36 37 defrosting conditions was greater in the cherries packed in 1 lb. polyethylene bags. Pretreatment of the pitted fruit with 802 in the concen- tration used prove to be very effective in the retardation of red color loss and browning. No 502 could be recovered in any of the SO2 treated fruit by direct acid hydrolysis (Moneir- Williams distillation method), or combination of saponifica- tion with sodium hydroxide and hydrolysis with acid. Experiments with addition of 802 just before defrosting to further improve red color retention under accelerated oxi- dation conditions showed that a concentration of 25 ppm 302 prevented red color destruction during 60 minutes of blending in the control samples, but did not prevent phenolic compounds oxidation, although the rate was half that without 302. Twelve ppm 50 practically prevented red color destruction and pheno- 2 lic compounds oxidation in the cherries pretreated with 1000 ppm SO prior to freezing. The addition of 25 ppm 302 bleached 2 the fruit. In conclusion 302 found to be effective in decreasing red color degradation and browning during frozen storage and subsequent defrosting. The concentration used did not impart an undesireable flavor or cause color bleaching. The results indicated that a slightly higher 802 concentration may be used for pretreating the fruit prior to freezing for better color retention. LITEHAT $2 C1T£D Baruch, P., and T. Swain. 1953. The effect of l-ascorbic acid on' the in-vitro activity of polyphenoloxidase from potato. Bioch. J. 55:392. I Bedford, C. L. 1965. Unpublished data and private communi- dation. Department of Food Science. Michigan State University, E. Lansing, Michigan. Bullis, D. E. and E. H. Wiegand. 1931- Bleaching and dyeing Royal Ann cherries for Maraschino or fruit salad use. 9335; State Coll. Agr. Expt. Sta. Bull. 275:3-29. Burton, H. S., and D. J. McWeeny. 1963. Non enzymic browning: The role of unsaturatéd carbonyl compounds as intermediates and of 802 as an inhibotor browning. J. of the Sci. of Food and Agr. 14:911. Diemeir, w. et a1. 1960. Influence of sulfurous acid and l-ascorbic acid in wine making. II. Inactivation of the polyphenoloxidase. Chem. Abst. (German) 55:5859 d. DuBois, C. w. 1949. Ascorbic acid and color in food products. Food Tech. 3:119. Goldstein. J. L., and T. Swain, 1963. Change in Tannins in ripening fruits. Phytochemistry. 2:371. Goodman, L. P. and P. Markakis.1965. Sulfur dioxide inhibition of anthocyanin degradation by phenolase. J. of Food Science 22:135-137. 2“) J7 i) t Grommeck, d. and P. Karkakis. 1964. The effe - cf peroxidase on anthocyanin pigments. J. Food Science 22:53-57. Guadagni, D. G., D. G. Sober, and J. S. Wilbur. 1949. Enzymatic oxidation of phenolic compounds in frozen peaches. Food_lech. 3:359-364. Guadagni, D. G., C. C. Nimmo, and E. F. Jansen. 1957. Time- Temperature tolerance of frozen foods. XI. Retail packs of frozen red sour pitted cherries. Food Tech. 12:36-40. Guadagni, D. G., J. Harris, and K. M. Eremia. 1963. Factors affecting quality of pies prepared from frozen bulk-pack red sour pitted cherries. Food Tech. 11:103-106. Hamburger, J. J. and M. A. Joslyn. 1941. Auto-oxidation of filtered citrus Juice. Food Research. 6:599. Hillis, w. E. and T. Swain. 1959. The phenolic constituents of Prunus domestica. J. of the Sci. of Food and Agr. 10:135. Huang, H. T. 1955. Decolorization of anthocyanins by fungal enzymes. J. Agr. Food Chem. 31:141-6. Ingraham, L. L. 1956. Effect of ascorbic acid on polyphenolo- xidase. J. Am. Chem. Soc. 18:5095. Johnson, G. and D. K. Johnson. 1952. Natural flavor retained in new frozen uncooked apple pulp. Food Tech. 6:242-5. Joslyn, M. A. and L. A. Hohl. 1948. The commercial freezing of fruit products. Uni. California Agr. Expt. Sta. Bull. 203:24. 43 Joslyn, M. A. and J. B. S. Braverman. 1954. The chemistry and technology of the pretreatment and preservation of fruit and vegetable products with 302 sulfites. Advance 1n Food Research. 5:97. Jurd, L. 1964. Reactions involved in sulfite bleaching of anthocyanins. J. Food Science 22:16. Krueger, H. C. 1950. The effect of ascorbic acid on the enzymatic oxidation of monohydric and o-dihydric pehnols. J. Am. Chem. Soc. 11:5582. LaBelle, a. L., J. c. Moyer, w. B. Robinson and D. B. Hand. 1953. Causes of scald in red tart cherries. Food Tech. 11:94- 98. Lee, F. A., w. A. Gortner, and J. Whitcombe. 1949. Effect of freezing rate on fruit. Food Tech. 3:164. Li, K. C. and A. C. Wagenknecht. 1956. The anthocyanin pig- ments of sour cherries. J. Am Chem. Soc. 18:979. Luther, H. G. and G. O. Cragwall. 1946. Ascorbic acid - citric acid prevent browning of cut fruits. Food Ind. 183690. Mason, H. S. 1957. Structure of melanins. P1gment cell biology by Gordon. Meschter, E. E. 1953. Effects of carbohydrates and other factors on strawberry products. J.4ggr. and Food Chem. 1:574. Peng, C. Y. and P. Markakis. 1963. Effect of phenolase on anthocyanins. Nature. 129:597-598. 41 Ponting, J. D. and G. Johnson. 1945. Determination of 302 in fruits. Ind. and Eng: Chem. analytical Ed. 11:682. Ponting, J. D. 1960. The control of enzymatic browning of fruits. Food Enzymes by H. w. Schultz. The Avi Publish- ing Company, Inc. pp. 105-123. Robinson, E. S. and J. M. Nelson. 1944. The tyrosine - tyrosinase reaction and aerobic plant reSpiration. Archives of Biochem.‘4:111. Rogers, A. J. 1940. How to quick freeze cherries success- fully. Refrigerating_§ng. 44:215. Rosenblat, m. and J. Peluso. 1941. Determination of Tannin: by Photocolorimeter. J. Assoc. official Agr. Chemists. g4: 170. Sastry, L. V., B. S. Bhatia and G. Lal. 1961. Studies on aspects of Custrad apple peroxidase. J. Food Sci. 16: 244-7. Sondheimer. E. and Z. I. Kertesz. 1948. Anthocyanin pigments. Anal. Chem. 14:245-8. . . 1952. The kinetics of the oxidation of strawberry anthocyanins by H202. F221 Research. 11:288. Schiner, D. M. 1960. The enzymic decolorization of antho- cyanin pigments. Cornell Uni., Ph.D. Thesis. Stadtman, E. R. 1948. Non-enzymatic browning in fruit pro- ducts. Advance in Food Research. 1:325. Starachan, C. C. and A. w. Moyls. 1949. Ascorbic, citric and dihydroxy maleic acids as antioxidants in frozen pack fruit. Food Tech. 3:327. 42 Tauber, H., I. S. Kleiner and D. Mishkind. 1935. Ascorbic acid (vitamin C) oxidase, J. Biol. Chem. 110:211. Wagenknecht, A. C., D. M. Scheiner and J. P. Van Buren. 1960. Anthocyanase activity and its possible relation to scald in sour cherries. Food Tech. 14:47-9. Weissberger, A. and J. E. LuValle. 1944. Oxidation process XVII. The autoxidation of ascorbic acid in the presence of Cu. J. Am. Chem. Soc. 66:700. Whittenberger, R. T. and C. H. Hillis. 1956. Bruising causes cherry discoloration. Canner and Freezer. 123:14. APPENDIX a) -4 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 mem.a .cas 000 000.0 000.0 000.0 00m.a 000.0 mao.H N00.0 00m.a 000.0 moo.a 000.0 000.0 000.0 000.0 mmm.o mmm.a .200 000 000.0 000.0 000.0 000.0 000.0 000.0 «00.0 00m.a 000.0 moo.H 000.0 000.0 000.0 mao.H N00.0 mmm.a .caa 000 000.0 m00.H 0m0.0 000.0 000.0 000.0 0m0.0 000.0 000.0 NNO.H mm0.0 me.H 000.0 Nmo.H 000.0 00m.a .2a0 000 sm\mam< am\man¢ em\mam¢ smxmama 10.0 ma . s.m may 0.m mg 10.0 me . 0.m mcv 0.m m0 moflmhmhmaa moflmhmhhdo 00000 Honooam .208 em .:He 0 0800 00000 medecoam oads .00:0000 400 00 00006000 00000 0.0 we 0:0 0.n m0 2003009 cosmpompav mm one :.m ma pd mocmnnowpm HoHoo cop no hpaadnmpm “a 00000 nu .00000020 000000 000w 000 as man 00 hoca9nom9<* 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 m00.0 000.0 000.0 00m.o, 000.0 000.0 000.0 .000 00 000.0 0m0.o 000.0 000.0 000.0 000.0 m00.o 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 .000 nu 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 .000 00 000.0 000.0 000.0 n00.o 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 .000 00 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 .000 on 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 000.0 .000 m0, 0w\n0n0 0w\00n< 00\m0n< 00\n000 0w\m0m< em\m0m< ew\m0m< 0m\m0n< 00000 00.0 000 000m 000 00.0 000r00nm 000,00.n 000 00.m 000 0o.m 000 «0.0 00V 000000 00 . 00 n0 0 00000 0000 000 00000 wC0020Hm 080B .o.m mm 000 0.m ma 000059 00000001830000 00 00000000 00000 m0dam> 00000 00A 00 h00a09w0m «N 00909 .000000HQ00 0 0o m0w000>0 000 005000 0:8 .m00m0000 00000 00000000 000000 00 05H0> 0oaoo 0000020 00 0000H00 00 0000adoamo mmoa & *# 00000030 000000 000w 000 0000050900 :8 man 00 0000900090 .2. 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