Effect of Carbon Dioxide and Sodium Benzoate on Vitamin C Content of Orange Juice AGNES
FAYAIORGAN,
I. LANGSTOK, AXD k W . 4 FIELD Universitv of California, Berkeley, Calif. CATHARIKE
T w o samples each qf California calencia and upon the iitamin C of orange juice. Pusieurized navel orange juices kept in frozen storage for 8 orange juice when bottled with carbonated water a1 to 18 months show antiscorbutic value f u l l y equiva- 2.5 kg. per sq. cm. pressure retains the full ititamin lent to that of freshly extracted orange juice. The C; isalue qf the fruit juice which it contains, 6 or 7 minimum amount required for .full protection qf cc. being approximately equiculent to I cc. qf freshly the sfandard guinea pig f o r 60 d a y is 1.5 or 2 cc. extracted orange juice. ,4similar preparation condaily. -Yo difference in vitamin C d u e between taining no excess carbon dioxide has lost a considvalencia and naz!el oranges is observed. -4 sweetened erable portion of its citamin activity. The same commercial orange juice prepared i n two forms (wilh difference in lesser degree is found in t u o correand without 0.1 per cent sodium benzoate) is tested sponding undiluted orange juices prepared by pasf o r antiscorbutic activity and found to be alike in teurization with and without carbon dioxide. T h e fhe two cases. This orangeade appears to h w e at relation qf fhis profectice action of the carbon dioxide least two-thirds the citamin C d u e of ahole fresh to the diminution i n amount of air retained i n the orange juice, a n y diminution being due no doubf juices is obcious and is further evidence of the wellio the dilution with sugar. Sodium benzoate in the known sensiiivity of the antiscorbutic vitamin to deamounts used appears to haite no deleferious efect sfrucfion by oxidation.
T
HE merit of fresh orange juice as a source of the antiscorbutic vitamin C has remained unquestioned since
that amount was required. Orange juice canned by heating to 100" C. for 5 minutes showed no loss when used immedithe work published in 1920 by Delf ( 2 ) and others in ately. -4test of the same product 4 years later showed a which by macroscopic methods of diagnosis 1.5 cc. were small loss of original value. The result when sulfite preservafound to be a minimum protective dose for guinea pigs. The tives were added to lemon juice was a loss of vitamin C question was a t once asked, however, whether the various amounting to five-sixths of the original value in 4.5 months. TKOarticles ( I , 9) have recently appeared in which the modern methods of concentration and preservation by which fruit juices are made available in the markets may not destroy vitamin C content of frozen orange juice has been reported in whole or in part this valuable property of most citrus fruits. as undiminished after 5 and 10 months of storage. Kelson Recent developments in the production of f i ozen orange juice and Mottern (9) further found that exclusion or expulsion of for retail distribution make this question even more pointed ail from the juice previous to rapid freezing had little effect than formerly. Numerous orange juice preparations now on upon vitamin preservation. Juice from Florida valencia the market, concentrated, diluted, sweetened, canned, bottled, oranges frozen under head space of oxygen was apparently carbonated, pasteurized, and preserved in a variety of ways, as rich in antiscorbutic value a6 that which had been extracted lay claim to health value presumably chiefly because of their and frozen under head qpace of nitrogen. Conn and Johnantiscorbutic property. It has seemed worth while t o test son (1) tested commercially prepared frozen juices made by comparative methods a few of the products made by these chiefly from Florida and Porto Rican valencia oranges by daily feeding to guinea pigs at 1.5- and 3-02. levels. Whole means. Givens and RScClugage (4) in 1919 demonstrated the prac- juice and depulped or rentrifuged and filtered juice were ticability of preparing highly concentrated orange juice in fed separately and were found t o be equivalent to each other solid form. Evaporation a t low temperature for a short and to fre3hly extracted orange juice as sources of vitamin C. The processes u'sed in the handling of the orange juices here period was shown to have little effect on the antiscorbutic properties, and the extracted residue retained considerable reported are: potency after 3 months of storage. A year later Harden and 1. Freezing and storage in the frozen condition. Robison (5) obtained tests proving dried orange juice to be 2. Addition of not more than 0.1 per cent sodium benzoate active after 2 years of storage. Hess and Unger (6) found to canned or bottled juice. 3. Carbonation-that is, treatment with carbon dioxide gas that aging in a refrigerator for some months caused a loss of under pressure previous to bottling and pasteurization. vitamin C in fresh orange juice. The later findings of Delf (3) summarized several imporTESTS ON FROZEN STORED ORANGEJUICE tant storage considerations. The acidity of fruit juices was Four samples of frozen juice were tested, two from valengiven as an influence promoting stability of the antiscorbutic property. No deterioration occurred in frozen orange juice cia and two from navel oranges, all grown in California. after 17 months of storage, a t which time 1.5 cc. fully pro- Sample -4was valencia juice which had been held in the frozen tected guinea pigs for 90 days. After 5 gears, however. twice condition for nearly a year before it was sent here and was 1174
ND i E N G I N EE R I N G C H E M I ST K Y I SDUSTR IAL J
October, 1933
about 18 months old when these tests were completed. Sample V, also made from valencia oranges, was freshly extracted frozen juice which had been held in storage from 2 to 8 months before the tests were finished. Sample B (from navel oranges, freshly extracted and held in frozen storage about 6 to 8 months during the tests) and sample C (also from navel oranges) were similarly held for about 3 months. A considerable proportion of pulp is retained in all these juices, whether designated as frozen whole juice, orangeade, or a diluted beverage. The chief interest in the testing of the frozen juices lies in the establishment of the stability of these products during t h e several months of frozen storage necessary for the completion of the biological tests. California tree-ripened frult was used for the preparatlon of the frozen juice. The oranges mere soaked, scrubbed by rotating brushes, cut by rotary knives, and either machine- or handreamed. This pulpy juice TYas then screened, removing threefourths of the pulp. The juice was frozen to a slush mthin an hour of reaming. The slushed juice was packed in paraffined paper cartons and hardened in a cold room at -12' C. -ill metal parts in contact with the juice were made of monel metal and stainless steel. The frozen juice was shipped to Berkeley, Calif., packed in solid carbon dioxide and kept during the test period at a temperature of -17" C. The basal diet used in the tests here reported nas a modification of that employed by Sherman, LaMer, and Campbell (IO) supplying all known factors in amounts necessary for normal growth and development, excepting vitamin C. 811 the orange juice doses were fed daily by pipet to the animals so that no uncertainty could ensue as to the ingestion of the full amounts allotted. As is indicated elsewhere (a), the 60-day experimental period has been found to yield as consistent and reliable data as the period of 90 days previously employed, and was therefore used in this experiment. As shown in Table I all four of these frozen juices retained t h e full antiscorbutic value of freshly expressed orange juice, designated sample VIII. A daily dose of 2 cc. appears to afford practically complete protection in all cases and in nearly the same degree as t h a t shown by the fresh juice. The probable correctness of 1.5 cc. as the minimum protective daily dose of fresh undiluted orange juice is confirmed. KO change in vitamin content appears to have taken place during t h e 18-month frozen storage period of sample A, again confirming t h e findings of Delf ( 3 ) . Part of sample A, which had been melted, packed in 8-ounce cans, and refrozen, was found t o yield the same protective value as the remainder of that sample. Certainly there can be no objection to the use of fiozen orange juice on the score of its vitamin content as compared with the fresh juice when the storage period does not exceed 18 months. It is probable that deterioration after t h a t time, if it occurs, is slight and gradual. KO tests have been made in this laboratory on juices held longer than 18 months. TABLE I. COMPARIsON AND
O F L4STISCORBGTIC VALUE FROZEN ORANGE JUICE
O F FRESH
G.rnr
STORAQESAM-
JTJICE PERIODPLE Months 8 V
Frozen valencia
1.5 2 3
3 8 10
DAY^
Grams Grams Grams Grams 319 490 464 2.4 328 468 451 2.0 326 489 466 2.3
SCoREb
2 2
0
A
2 3 4
6 6 8
346 357 333
520 549 576
607 518 574
2.7 2.7 4.0
1 2 0
3
C
1 2 3
9 7 13
315 323 318
396 434 496
319 405 473
0.0 1.4 2.7
0-1 0
2 3 4
4 5
349 322 322
499 544 477
486 524 471
2.3 3 3 2.5
1 1 0
ExVI11 1 13 366 462 423 1.4 pressed 2 12 350 496 486 2.2 daily 3 16 342 499 499 2.6 For 60 days. On a scale of increasing severity of scurvy symptoms, 0 to 24.
5 1 0
B Fresh
b
cc .
IN KT. PER SCERYY
18
Frozen navel
a
Av. WEIQHTOF FED 4x1ANIMALS DAILYMALS Initial Max. Final
MOUNT
6
6
1175
The experience of Koch and Koch (7) with loss of antiscorbutic potency by commercially expressed orange juice when stored frozen or a t refrigerator temperature is a t variance with results obtained here and with those of Delf ( S ) , Nelson and Rlottern (9), and Conn and Johnson ( I ) . Koch and Koch note also a curious preservative effect of the added citric acid and sugar in a somewhat diluted pulpy juice; this effect is lost on further dilution to 12.3 times the original volume.
TESTSOK ORAKGEPRODUCTS PRESERVED WITH SODIUM BEKZOATE The product studied in the attempt to discover the effect of sodium benzoate upon the vitamin was a swedened orange juice similar to a number of preparations which have been on the market for some time. The screened orange juice prepared as previously described was mixed in glass-lined tanks Tyith 40 per cent of its weight in cane sugar and 4 per cent of a concentrated lemon juice. To this mixture was added sodium benzoate in amount such that the finished sirup contained less than 0.1 per cent. Quart bottles were then filled with the sirup and heated in a continuous pasteurizer for 25 minutes a t a temperature of 63" C. in the bottles, then cooled a t once. The time elapsed betiyeen reaming and bottling was from 2 to 3 hours. All of these samples were kept a t 0" C. during the testing period. T.4BLE
P E R CENT s O D I U M BENZO.4TE A4STISCORBUTICP R O P E R T Y O F ORkKGE ,JUICE
11. EFFECT O F 0.1 THE
CPON
CHANQE I N DuR&AMOUNT Av. WEIQETOF BODYWT. TIOX OF FED ANIANIMALS PER FEEDSCURYY JUICE DAILY MALS Initial M a x . Final DAY INQ SCORE Cc Grams Grams Grams Grams Days None 12 339 358 227 -3 6 29 17 +2.2 498 469 58 2 3 17 338 Without so+1.2 450 420 dium ben- 4 11 348 60 1-2 +2.9 519 513 60 0 340 zoatea 5 14
.
485 +3.3 349 493 K i t h so3 14 60 2 460 +2.1 57 0-1 337 489 dium ben- 4 17 324 495 soateb 5 18 +2 8 58 0 490 a Valencia uhole juice, largely depulped, containing 40 per cent cane sugar, uithout sodium benzoate. b Same, a i t h 0 1 per cent sodium benzoate.
The sirup was supplied by the manufacturer in two forms, one sample having no added benzoate and the other containing not more than 0.1 per cent sodium benzoate, all other manipulation being the same. Both types were used in dosages of 3, 4, and 5 cc. daily for the usual period of 60 days. 3 s shown in Table 11. 3 cc. of either form-that is, with or without sodium benzoate-provided practically as complete protection from scurvy as was afforded by 2 cc. of fresh or frozen whole orange juice. This decrease of the original antiscorbutic value is probably due t o the dilution of the juice by cane sugar added in the packing process. The frozen valencia juice (sample V) and frozen navel juice (sample C) reported in Table I were furnished as representing the unfrozen juice from which the diluted products were made. There was available therefore a reliable measure of the amount of loss of antiscorbutic value through dilution by addition of sugar. It is obvious that sodium benzoate in the amount used in these products has no effect upon the vitamin C of the orange juice. Williams and Corran (11) using one animal on each of four doses, consisting of 1 to 2.5 cc. of lemon juice preserved with 0.05 per cent sodium benzoate, found only negative results and concluded that the benzoate was neither a good preservative of the vitamin nor of the gross palatability of the juice. However, their sample was fermented, had been kept some time, and was of unknown original vitamin content. It is a fact not always borne in mind by those who use or prescribe orange juice for its antiscorbutic value, particularly for infants, that the sweetened orange juice products or
INDUSTRIAL AND ENGINEERING CHEMISTRY
1176
VOl. 25, No. 10
orangeades, of which the one here tested is an excellent example, are already somewhat diluted as t o juice content even though concentrated as to sugar content by addition of cane sugar. These beverages are not vitamin concentrates, even though their sugar contents make i t desirable to dilute them before use. It is reassuring, however, that they may contain the full antiscorbutic potency of their orange juice content, in this case about two-thirds of the product.
the carbon dioxide treatment is the only factor of difference between the two samples, this must be considered t h e protecting substance in the better preservation of the vitamin evinced by sample I. Apparently, complete retention of a n antiscorbutic content equivalent t o that of the original whole juice, designated sample V, is shown by sample I, since, although containing a maximum of 18 per cent of orange juice, it evidenced the antiscorbutic effect of a t least 18 per cent of t h e vitamin C of the whole juice (sample V) from which EFFECTOF CARBONDIOXIDEIN DILUTEDORANGEJUICE i t was made. On the other hand, sample I1 which was no A study of the effect of carbon dioxide upon the preserva- more diluted than sample I had less than 10 per cent of the tion of vitamin C was undertaken by testing five samples of original vitamin content of t h e whole juice, sample V. Sample VI was equivalent in antiscorbutic value to V, thus orange juice products prepared as follows: indicating complete preservation of the vitamin when the I. Slightly sweetened valencia orange juice, diluted with juice was pasteurized and carbonated. But sample VI1 apwater containing 0.1 per cent dissolved carbon dioxide. To pre- peared to be somewhat less potent than V or VI, indicating a pare this, the sirup, used in making the sample described in the previous section, without sodium benzoate, was diluted with possible loss of antiscorbutic activity due to pasteurization filtered carbonated water at a pressure of 35 pounds per square and storage without the benefit of carbonation. Doses inch (2.5 kg. per sq. cm.) in the proportion of 2.5 ounces of sirup ( 2 cc.) of VI, which provide practically complete protection, made up to 12 ounces in black bottles. The product was then in VI1 are obviously insufficient for similar protection. pasteurized for 25 minutes a t 63" C. and cooled. It is not surprising that carbon dioxide treatment should 11. Juice exactly similar to sample I, diluted with water conhave this protective effect if the sensitiveness of vitamin C taining no added carbon dioxide. V. Frozen, untreated, whole valencia orange juice, similar to oxidation is recalled. Smaller pressures of air or oxygen to that from which samples I and I1 were made and the same as can be exerted by juices charged with carbon dioxide, and sample V of Table I. VI. Frozen orange juice, similar to sample V, packed in slower rates of oxidation may result, This is in line with the bottles, but pasteurized for 25 minutes a t 63" C. and treated report of Zilva (12) in which lemon juice heated for 2 hours with carbon dioxide at a pressure of 2.5 kg. (undiluted sample I). in an atmosphere of carbon dioxide was found to have reVII. Frozen orange juice pasteurized as sample VI but with- tained its full vitamin C value. out carbon dioxide treatment (undiluted sample 11). Samples V, VI, and VI1 thus represent 100 per cent orange juice, and samples I and I1 represent dilutions containing 14 to 18 per cent of the same orange juice. All samples were kept in frozen storage throughout the testing period of 8 months. TABLE111. ANTISCORBUTIC PROPERTY OF ORANQE JUICE PREPARATIONS AS AFFECTED BY CARBON DIOXIDE AMOUNT
Av. WEIGHTOF FED ANIANIMALS SAMPLED.AILYM A L S Initial N a x . Final CC Grams Grams Grams I 2-6 19 375 332 256 7 6 341 461 407 8 8 333 480 457 9 7 468 453 325
.
CHANQE I N DURABODY TION OF WT. PER FEED-SCURVY DAY I N Q SCORE Grams Dags -1.5 49 11 $0.9 55 4 +2.1 60 3 $2.1 60 2
ACKKOWLEDGMEKT Samples V and C were provided by the Mission Dry Corporation, formerly the California Crushed Fruit Corporation of Los Angeles, to whom t h e authors are also indebted for the orangeade preparation, furnished with and without sodium benzoate (Table 11) and for the carbonated and other juices (Table 111). The hlission Dry Corporation also prepared samples I, 11, V, VI, and VI1 for these tests. Sample I in the series of orange products here described is identical with their orange juice beverage called "hlission Orange Dry." LITERATURE CITED
I1
3-6 10 12 13
10 3 6 2
333 345 338 363
371 382 435 464
200 254 390 401
-3.9 -2.4 $0.9 +0.6
34 42 59 59
I6 11 6 9
(1) Conn, L. W., and Johnson, A. H., ISD. ESG. CHEM.,25, 2 1 8 (1933). (2) Dell, E. hl., Biochem. J . , 14, 211 (1920). (3) Ibid., 19, 1 4 1 (1925).
V
1.5 2 3
3 8 10
319 328 326
490 468 489
464 451 466
$2.4 $2.0 $2.3
60 60 60
2 2
1.5 2 3
4 9 7
353 331 343
482 466 514
482 464 497
$2.1 +2.2 +2.5
61 60 60
1 1 0
2 3 4
7 11 3
321 332 354
378 484 510
318 475 462
-0.1 +2.4 +1.8
55 60 60
7 1 3
(4) Givens, M.H., and McClugage, H. B., Am. J . Diseases ChiG dren, 18, 30 (1919). (5) Harden, A., and Robison, R., Biochem. J., 14, 1 7 1 (1920). (6) Hess, A. F., and Unger, L. J., J . Bid. Chem., 35, 479 (1918). and Koch, F.C., IND.ENG.CHEM.,24, 351 (1932). (7) Koch, E. &'I., (8) Morgan, A. F., Field, A, and Nichols, P. F., J. Agr. Research, 42, 35 (1931). (9) Nelson, E. R'I., and Mottern, H. H., ISD.ESQ. CHEX.,25, 216
VI VI1
0
As may be seen in Table 111,sample I provided nearly complete protection from scurvy when 8- or 9-cc. daily doses were given, and, when 6- or 7-cc. doses were used, as good protection as was afforded b y 12 or 13 cc. of sample 11. Since
(1933). (10) Sherman, H. C., La Mer, V. K., and Campbell, H., J . Am. Chem. Soc., 44, 165 (1922). (11) Williams, J., and Corran, J. W., Biochem. J., 24, 3 7 (1930). (12) Zilva, 9. S., I b i d . , 16, 4 2 (1922). RECEIVED May 2, 1932; resubmitted April 4, 1933.
There are a few towns, however, ivhere liquid chlorine WATERTREATMENT IN QUEBECASD MARITIME P R O ~ I X C E provinces. S. Public water supplies in the Provinces of Quebec, New Brunswick, is used. In the Province of New Brunswick, Fredericton is the Nova Scotia, and Prince Edward Island are soft and quite pure, only city w t h a filtration plant. It uses about 70,000 pounds and there are comparatively feT7 filtration plants other than sand of alum annually. The water is also chlorinated at all times; filters. In the large majority of cases the water is brought from about 6400 pounds of chlorinated lime are used each year. The nearby rivers or mountain lakes and stored in reservoirs. Some filters used in the Province of Quebec for the treatment of water smaller towns have artesian wells. The Public Works Depart- supplies are of the mechanical gravity type requiring the use of ments of Nova Scotia and Prince Edward Island state that there alum and chlorine. Some water supplies, including that of are no municipal water-filtration systems in operation in their Montreal, are treated by chlorine only.
