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Age-Thickening and Gelation of Sterilized Evaporated Milk 1

V. R. H A R W A L K A R , D. C. BECKETT, R. C. McKELLAR, D. B. EMMONS Food Research Institute Research Branch, Agriculture Canada Ottawa, Ontario, Canada K1A 0C6 G. E. D O Y L E Perfection Foods Ltd. Charlottetown, Prince Edward Island, Canada C1A 7M8

ABSTRACT

INTRODUCTION

Evaporated milk containing 26% total solids (15 batches) and evaporated skim milk containing 18% total solids (1 batch) were held at 4C before canning and sterilization. Sterilized samples were stored at 28C and examined periodically for 1 yr for changes in relative viscosity, gelation, granulation, and fat separation. Samples sterilized without cold storage remained liquid for 12 too. As time of cold storage of concentrates before steriiization increased, this tendency to thicken or gel during storage increased. Ten of 15 batches of evaporated milk cold stored for 3 days before sterilization had gelled after 12 mo along with 6 that had been held for 2 days. Samples stored cold for 3 days that did not gel after 12 mo had thickened more than controls. Samples that gelled contained no microorganisms capable of growth at 30C and showed no appreciable change in proteolysis or pH. Heating samples at 55 to 60C for 30 rain after 3 da~cs cold storage,and then cooling to 20-C (so-called anti-coolaging treatment") before canning and sterilization did not prevent gelation or increase in viscosity during storage. Oxidizing conditions such as aeration and peroxide treatments accelerated whereas reducing conditions such as antioxidant treatments tended to delay but not prevent age-thickening and gelation.

Evaporated milk prepared by conventional sterilizing methods usually remains free from age-thickening or gelation after long storage at ambient temperatures (over 12 too). However, sporadic appearance of gelation in some evaporated milk samples has caused serious problems for some manufacturers. The cause of this defect is not apparent and has not been related to any of the customary processing variables such as time and temperature of forewarming, homogenizing pressure, added orthophosphates, etc. However, most batches that showed age-thickening or gelation in a particular dairy plant in Canada were prepared from concentrates held in cold storage before canning and sterilization. Similar observations were made by Dutch workers (3), who observed gelation in samples of evaporated milk and demonstrated a relationship between gelation and cold storage of concentrates before sterilization. They demonstrated this relationship only in evaporated milks conforming to European standards (fat/ SNF (solids-not-fat) of 9/22 or 10/23). In their experience, evaporated milk made to North American standards (fat/SNF of 8/18) did not show this defect. In view of the similarity of the problem described by the Dutch workers and the agethickening described, our first objective was to see if the phenomenon described by the Dutch workers could be observed in evaporated milk made to North American standards. The second objective was to find suitable explanation for the age-thickening or gelation in evaporated milk.
MATERIALS AND METHODS

Received March 4, 1982. :Contribution No. 494 from the Food Research Institute. 1983 J Dairy Sci. 66:735-742

Evaporated milk was prepared in an evaporated milk plant by a conventional method. 735

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HARWALKAR ET AL. ward r o t a t i o n and an average d e t e r m i n e d after the two top and b o t t o m readings were omitted. T h e average arbitrarily was taken as relative viscosity of the sample; a relative viscosity o f 10 is 166 centipoise. Granulation was observed in thin films of sample drawn with a wire loop (2 cm i.d.). Fat separation and gelation were assessed by visual observation. Standard plate c o u n t (SPC) was as described in Standard Metbods for tbe Examination of Dairy Products (6) with the e x c e p t i o n that dilutions were with .1% peptone to facilitate recovery of organisms (7). Incubation was for 48 h at 32C. Degree of proteolysis was measured as the increase in trichloroacetic acid-soluble free amino groups (5).
RESULTS A N D DISCUSSION Viscosity and Gelation

Briefly, raw milk was f o r e w a r m e d to 8 8 C and held in a " h o t well" for 25 min. A hot a q u e o u s slurry of stabilizing salt (disodium phosphate) and carrageenan were p u m p e d into the hot well. The a m o u n t of stabilizing salt and carrageenan added were usually 11.3 and .68 kg per batch of 3.17 x 104 kg raw milk. The milk then was c o n c e n t r a t e d in a double-effect evaporator, first effect at 68.3C with 57.6 kPa of vacuum and in the second effect at 43 to 45C with 95 kPa of vacuum. The concentrate then was h o m o g e n i z e d at 22.75 and 4.8 MPa at 43C, cooled to 6.7C, and p u m p e d to a holding tank for standardization to 8% fat and 26% total solids. A similar procedure was used for preparing evaporated skim milk. A portion of the c o n c e n t r a t e was held at 4C in large (100 liter) plastic containers for varying times. Some of this concentrate was treated specially as described later. Following this holding period and special t r e a t m e n t , the c o n c e n t r a t e was canned and sterilized at 119C for 10 min in an FMC continuous rotary sterilizer. Cans were cooled and stored undisturbed at ~ 2 8 C . The stored samples of evaporated milk were e x a m i n e d periodically for changes in relative viscosity, granulation, fat separation, and gelation. Viscosity o f the samples was measured at 2 1 C by a Brookfield viscometer m o u n t e d o , a Helipath stand with the spindle " A " at 12 rpm. Readings were taken after every down-

Data in Table 1 show the effect of cold storage of mitk c o n c e n t r a t e before sterilization on the relative viscosity during storage of samples of c o n c e n t r a t e d milk f r o m 15 batches. The upper p o r t i o n of Table 1 shows the mean relative viscosity of ungelled samples; gelled samples (number in brackets) were excluded. Because the high viscosities of gelled samples are not included, some t r e a t m e n t s with low

TABLE 1. Effect of cold storage of milk concentrates before sterilization upon changes in relative viscosity during storage. Treatment (days of cold storage at 4C) 0 1 2 3 0 1 2 3 Relative viscositya after storage (too) (Mean of 15 batches) 0 6.5 6.2 7.4 9.7 8.0 7.9 9.3 9.8 3 8.0 9.6 12.9 21.8 7.4 9.0 10.8 12.2 6 15.5 23.0 33.0 (2) 50.0(6) 10.9 12.0 23.0 34.6 9 21.9 22.1 (2) 48.0 (4) 50.0 (9) 11.8 17.6 38.0 48.1 12 25.9 32.0 (2) 37.0 (6) 49.0 (10) 12.4 21.0 33.4 48.6

Sample All 15 batches of evaporated milk

Ungelled evaporated milkb

aRelative viscosity data are means of the ungelled samples and the number of gelled samples is indicated in brackets. bData are for the five batches that did not gel at 12 mo in the 3-day cold-storage treatment. Journal of Dairy Science Vol. 66, No. 4, :1.983

GELATION OF EVAPORATEDMILK gelation (e.g., 2-day cold storage after 9 mo) show viscosities as high as another treatment with frequent gelation (e.g., 3-day cold storage after 12 too). A better comparison is on the five samples for which no gelation occurred during 12 mo. These are in the bottom of Table 1. Viscosity of evaporated milk increased during storage. In the 15 samples receiving no cold storage treatment, relative viscosity increased from 6.5 to 25.9. In the five samples that did not gel, relative viscosity of samples with zero cold storage increased from 8.0 to 12.4 and that of samples with 3 days of cold storage increased from 9.8 to 48.6. It is normal for evaporated milk to show a decrease in viscosity after manufacture, after which it increases. This was marginally apparent in Table 1 in the five samples that did not gel; relative viscosity at 3 mo was 7.4 compared to 8.0 at 0 and 10.9 at 6 mo. Viscosity had decreased and then increased sufficiently in the 15 samples that the mean relative viscosity at 3 mo was 8.0, compared with 6.5 at 0 time and 15.5 at 6 mo. Cold storage of concentrate before sterilization markedly influenced the relative viscosity, during storage. The 15 lots sterilized without cold storage remained liquid for over 12 mo of storage. Viscosity of samples stored cold before sterilization showed greater increased in relative viscosity and gelation during subsequent storage, the increase being proportional to the period of cold storage. At 12 mo, the mean relative viscosity with 0-clay cold storage treatment was 25.9. However, two of the 15 samples cold-stored for 1 day, 6 of the 15 samples coldstored for 2 days, and 10 of the 15 samples cold stored for 3 days had gelled. At shorter

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storage times, viscosity and frequency of gelation increased. Even after 3 mo storage there was a regular increase in viscosity as cold storage treatment increased from 0 to 3 days. In the five samples that remained ungelled after 12 too, the mean relative viscosity had increased from 12.4 to 48.6 as the time of cold storage before sterilization increased from 0 to 3 days. There were marked differences among lots in the way in which cold storage period affected viscosity and gelation during subsequent storage of the evaporated milk. It was typical that viscosity increased with time of storage and with time of cold storage before sterilization. Thus, susceptible lots gelled quickly with 3 days of cold storage but less quickly with 1 day cold storage. Resistant samples did not gel but showed typical increases of viscosity. Age-thickening also was accelerated when evaporated skim milk was cold-stored before sterilization (Table 2). At 6 too, the sample cold-stored for 3 days showed a substantial increase of viscosity and presumably would have gelled sooner than others. Evaporated skim milks were more susceptible to gelation; they gelled between 6 and 9 mo of storage.
Granulation

Granulation appeared during storage as samples thickened. These samples usually did not show any granulation in the freshly sterilized samples. The appearance of granulation usually preceded thickening or gelation during storage. The frequency of appearance of granulation during storage increased with the time of cold storage of the unsterilized concentrate (Table 3). There were some samples in which granula-

TABLE 2. Effect of cold storage of skim milk concentrate before sterilization upon changes in relative viscosity during storage. Days of cold storage at 4 C 0 1 2 3 Relative viscosity after storage (rno) 6 9 3.9 4.0 7.8 31.2 Gelled Gelled Gelled Gelled

0 2.5 2.4 3.0 2.9

3 4.0 3.7 3.9 3.8

12 Gelled Gelled Gelled Gelled

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HARWALKAR ET AL. stable proteases during cold storage, 4) coldinduced alteration of casein micelles, and oxidation/reduction reactions.
Quality of Raw Milk

tion was observed in freshly sterilized samples. These samples showed increased tendency to thicken or gel during storage. However, lack of sufficient number of such samples precludes a conclusive observation.
Fat Separation

There was little or no visual fat separation in any of the untreated control or treated samples. Therefore, thickening or gelation of evaporated milk was not a result of any unusual fat separation. However, samples of evaporated milk that received cold storage treatment showed after 6-mo storage more thickening at the top of the can than at the bottom whereas gelation of evaporated skim milk was accompanied by excessive synerisis, and the gelled portion settled to the bottom of the can. Thus, greater thickening at the top of the can is associated with fat or fat protein complexes. We confirm the observation of Dutch workers that gelation of evaporated milk is caused by cold storage of concentrate before sterilization. In contrast to their observations, we demonstrated this effect in evaporated milk made to North American standards (lower total solids than European evaporated milk). No suitable explanation is offered for this phenomenon of age thickening or gelation of evaporated milk. An explanation of the unusual gelation phenomenon was sought by examination of the possible involvement of 1) microbial quality of raw milk, 2) seasonal effects, 3) microbial growth and production and heat

The microbial quality of the raw milk used for preparing the evaporated milk did not seem related to age thickening or gelation. The raw milk used for the 15 batches varied from 70 103 to 385 103 cfu/ml (Table 4). Regardless of the initial count of the raw milk, evaporated milk thickened or gelled only when the unsterilized concentrates were subjected to cold storage. Corresponding batches of evaporated milk without cold storage remained liquid. Furthermore, some of the batches with higher initial count of raw milk merely had thickened whereas others with lower initial counts gelled within 12 mo.
Seasonal Effect

In commercial practice the problem of age thickening and gelation tends to be seasonal, more frequent in the early spring. This study tended to confirm this. Of the six batches processed between February 22nd and March 10th, 1980, only one batch that was cold-stored for 3 days had gelled by 12 mo. Of the nine batches processed between March 14th and May 12th, by 12 mo all batches with 3-day cold storage and seven with 2-days cold storage had gelled. By 6 mo five of the nine batches with 2days cold storage had gelled. These nine batches

TABLE 3. Effect of cold storage by unsterilized concentrate upon granulation and gelation during storage of sterilized evaporated milk. Days of cold storage at 4 C 0 1 2 3 0 1 2 3 Frequency of granulation and gelation a during storage (mo) 3 6 9 0 2 4 9 0 0 2 3 0 4 7 (2) 8 (6) 0 0 3 4 0 2 (2) 8 (4) 5 (9) 0 1 4 4

Sample All 15 batches of evaporated milk

0 0 0 0 5 0 0 0 1

12 3 5 (2) 6 (6) 4 (10) 1 2 4 4

Five batches of evaporated milk that did not gel at 12 mo

aThe additional number of batches that had gelled are shown in brackets. Journal of Dairy Science Vol. 66, No. 4, 1983

GELATION OF EVAPORATED MILK with 0- or 1-day cold storage showed more age thickening than the corresponding samples from the earlier six batches. Thus, seasonal factors, although influencing the susceptibility of the evaporated milk to age thickening, also accentuated the effect of cold storage upon gelation.
Microbial Growth

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During cold storage of the concentrate, psychrotrophs may have grown and produced heat-stable proteases, which acted slowly during storage and caused gelation as in some ultrahigh temperature (UHT) sterilized milk products (1, 4). This possibility seems unlikely for the following reasons, a) The increase in standard plate count (SPC) was usually slight after 3 days of cold storage of the concentrates (<100,000 cfu/ml), b) There was no apparent relationship between increase in SPC and age thickening or getation (Table 4). There were a few samples in which the SPC increased to over a million, yet the viscosity change was less than in samples with much lower increases in SPC. c) The pH at the end of 12 mo in thickened or gelled samples was similar to that in those which did not thicken (Table 4). d) The age-thickened or gelled samples showed no microorganisms capable of growth, e) There

was no difference of proteolysis in gelled and ungelled samples as measured by TCA-soluble free amino groups. For example, the mean of amino groups in eight lots of gelled or thickened samples, cold-stored for days before sterilizing, was 1.31/gmoles/ml; the mean in the eight comparable liquid lots of samples coldstored for 0 days was 1.40 #moles/ml. They had been stored an additional 2 yr at 20C after removed from t h e ' 2 8 C storage. Comparable amino groups in fresh and stored UHT milk were .7 and 1.6/amoles/ml (unpublished data); gelation in the latter was believed to be due to heat-stable proteases. Means for evaporated milk are double those of fresh UHT milk, as would be expected from the higher concentration of milk solids.
Cold-Induced Changes in Casein Micelles

Some of the changes in casein micelles resulting from cold storage are reversed by heating (8). For example, cold storage of milk delays rennet coagulation, but heating the milk at 55 to 60C (anti-cool-aging treatment) restores rennet coagulation time. Would a similar heat treatment of cold-stored evaporated milk affect the observed age-thickening or gelation? Data in Table 5 show that such heat treatment was ineffective in preventing gela-

TABLE 4. Relation between quality of raw milk, bacterial growth during 3-day cold storage of unsterilized concentrate, and viscosity change and pH of the sterilized concentrate at 12-mo storage. Standard plate count (cfu/ml) Batch no. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Concentrate Raw milk 135,000 250,000 280,000 360,000 280,000 340,000 360,000 282,000 160,000 385,000 290,000 268,000 135,000 235,000 70,000 0 day 132,000 47,000 135,000 21,000 5,000 15,000 8,000 65,000 105,000 85,000 16,000 1,000 16,000 97,000 37,000 3 day 1,560,000 39,000 411,000 6,000 7,000 47,000 63,000 210,000 210,000 210,000 130,000 37,000 53,000 TNTC 59,000 Relative viscosity of 3-day sample (after months) 0 12 9.2 6.5 31.9 14.4 13.0 5.7 15.3 5.5 6.0 5.1 5.5 6.2 5.6 9.5 5.9 32.0 21.0 Gelled 41.0 75.0 74.0 Gelled Gelled Gelled Gelled Gelled Gelled Gelled Gelled Gelled pH after 12 mo 6.05 ... 6.04 6.06 6.05 6.02 6.02 6.08 6.06 6.08 6.07 6.06 6.05 ... 6.08

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HARWALKAR ET AL. first applying vacuum (38.3 kPa for 20 min) followed by bleeding in nitrogen and bubbling for 20 rain. Incorporation of antioxidants was achieved by stirring in .02% antioxidant in milk concentrates at 55 to 60C. The two antioxidants were butylated hydroxytoluene (BHT) and propyl gallate. The above samples treated to accentuate oxidation or reduction were transferred manually to individual cans, sealed, and sterilized as described. Changes in viscosity and gelation during storage of these samples are in Table 6. Samples 1 and 2 are controls with holding of concentrates for 0 and 3 days at 4 0 Samples 3 and 4 show the effect of aeration. Aeration of the concentrate without cold storage showed slight thickening (compare samples 3 and 1). The aerated sample held 3 days showed no more thickening than the unaerated control (compare samples 4 and 2). Deaeration of the samples held 3 days did not affect viscosity changes (compare samples 7 and 2). Treatment with hydrogen peroxide of samples held both 0 and 3-day caused marked thickening or gelation compared to their controls (samples 5 and 6 vs. 1 and 2). Holding for 3 days before sterilizing markedly accentuated the effect of

tion. Therefore, physicochemical changes of the micelles leading to storage gelation must be of a different nature from changes affecting rennet coagulation time.

Oxidation-Reduction

Cold storage of milk concentrate may" increase dissolved oxygen, which in turn may influence gelation by oxidation reactions. This possibility was tested by treating milk concentrates to accentuate or to reduce oxidizing conditions. The unsterilized concentrates were treated as follows. To accentuate oxidizing conditions: a) concentrate, before or after cold storage, was aerated by bubbling air through it for 5 rain. The air flow was controlled to minimize excessive foaming, b) Hydrogen peroxide (H202) (.1%) was incorporated into the concentrate before sterilization. The incorporation of H202 in milk concentrates caused destabilization by normal sterilizing treatment (119C for 10 min). The sterilizing temperature, therefore, was reduced to 115C. This temperature in the presence of H202 was adequate to sterilize the sample. No such lowering of sterilizing temperature was necessary for skim milk concentrates with added H 2 0 ,.. These samples were stable under normal sterilizing conditions. The reason for this difference of stability of milk and skim milk concentrates in the presence of H202 is not known. Reducing conditions were introduced by deaeration and incorporation of antioxidants. Deaeration of concentrates was achieved by

the H202.
The presence of antioxidants appeared to delay age-thickening (samples 8 to 11). In sampies 8 and 10 the antioxidants were added to the 3-day-old concentrate. In samples 9 and 11 the antioxidant was added to the concentrate on 0 day and canned. The canned concentrates, then were held for 3 days at 4C before sterilization. There was little difference between sam-

TABLE 5, Effect of anticool-aging treatment a of cold-stored milk concentrates before sterilization upon age thickening, Days of cold storage at 4 C 0 3 3 Relative viscosity b after storage (mo) 0 6.7 8.8 13.8 3 8.5 14.9 27.8 6 11.4 38.6 54.2 (3) 9 13.1 50.5 (3) Gelled (5) 12 13.0 53.0 (3) Gelled (5)

Treatment ... .,. Anticool aging

Replicates 5 5 5

aCold-stored samples were heated at 55 to 60C for 30 min and were cooled to -20C before canning and sterilization. bThe viscosity data are means of the ungelled samples, and the number of gelled samples is indicated in brackets. Journal of Dairy Science Vol. 66, No. 4, 1983

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TABLE 6. Effect of oxidizing and reducing treatments of cold-stored milk concentrates before sterilization upon age thickening. Days of storage No. 1 2 3 4 5 6 7 8 9 10 11 at 4C 0 3 0 3 0 3 3 3 0/3 3 0/3 Treatment a ... Acetate Acetate H202 H202 Deaerate BHT c BHT d Propyl c gallate Propyl d gallate 0 6.3 9.5 11.4 9.8 5.4 6.1 11.4 7.8 9.3 8.8 9.9 3 8.2 14.8 16.1 15.3 7.1 48.6 16.1 14.0 13.1 14.6 14.7

Relative viscosityb after storage (mo) 6 10.5 38.6(1) 15.2 34.9 21.0 Gelled (4) 46.3 37.5 (1) 26.8 (1) 38.5 (1) 31.5 (1) 9 12.4 50.0(2) 33.0 39.0 48.0 Gelled (4) 49.5 39.0 (2) 34.0 (1) 29.0 (2) 18.0 (2) 12 9.8 49.0(2) 33.5 47.0 (2) 36.0 (1) Gelled (4) 56.5 (2) 45.0 (2) 38.0 (2) 43.0 (2) 33.0 (2)

aAll treatments had four replicates except 5, which had only three replicates. bThe relative viscosity data are means of ungelled samples, and the number of gelled samples is indicated in brackets.
C

Antmx~dant was incorporated after the 3-day cold storage.

dAntioxidant was incorporated before the 3-day cold storage.

pies to w h i c h a n t i o x i d a n t was a d d e d either b e f o r e or after cold storage; the a d d i t i o n o f a n t i o x i d a n t b e f o r e cold storage s e e m e d to be slightly m o r e beneficial. Evaluation o f the effect o f a n t i o x i d a n t s is c o m p l i c a t e d b y the fact t h a t e v a p o r a t e d milks vary in their susceptibility to gelation. Even t h o u g h t h e viscosity o f the less susceptible b a t c h e s s h o w e d less viscosity increases u n d e r r e d u c i n g c o n d i t i o n s , the n u m b e r o f susceptible s a m p l e s t h a t had geIled was t h e same. T h e r e f o r e , definitive c o n c l u s i o n s regarding t h e positive e f f e c t o f reducing condi-

t i o n s u p o n age-gelation are n o t possible. T h e e f f e c t of anti-cool-aging, h y d r o g e n p e r o x i d e , and a n t i o x i d a n t t r e a t m e n t on evap o r a t e d skim milk is in Table 7. T h e e f f e c t o f these t r e a t m e n t s was similar to the e f f e c t o n e v a p o r a t e d milk. However, skim milk samples were m o r e s u s c e p t i b l e to gelation. A p p a r e n t l y , fat had a p r o t e c t i v e e f f e c t against age t h i c k e n ing or gelation, p r e s u m a b l y b y i n t e r a c t i o n w i t h casein micelles. The ability of h y d r o g e n p e r o x i d e to augm e n t the gelation process is a d e f i n i t e indica-

TABLE 7. Effect of anticool aging, hydrogen peroxide, and antioxidant treatments of skim milk concentrates cold-stored before sterilization upon age thickening. Days of cold storage at 4 C 0 3 0 3 3 3 Relative viscosity after storage (too) Treatment . .. .._ H202 H202 Propyl gallate Anticool aging 0 2.5 2.9 2.8 3.1 2.8 3.0 3 4.0 3.8 4.5 Gelled 4.0 3.8 6 3.9 31.2 Gelled Gelled 27.6 25.5 9 Gelled Gelled Gelled Gelled Gelled Gelled 12 Gelled Gelled Gelled Gelled Gelled Gelled

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HARWALKAR ET AL. for his help in preparing the samples and D. Sibbitt for analysis of the samples.
REFERENCES

tion that o x i d a t i o n reactions are i m p o r t a n t in gelation. I n c o r p o r a t i o n of hydrogen p e r o x i d e also hastened gelation of UHT-sterilized evaporated skim milk (2). The antioxidants were not effective against age thickening or gelation; only slight protect i o n was observed. The apparent inability of the antioxidants to prevent gelation could have resulted f r o m p o o r dispersal of antioxidants. Alternatively, changes other than o x i d a t i o n m i g h t be involved. Changes during cold storage of evaporated milk b e f o r e sterilization seem to predispose the micellar system of the concentrates to age thickening or gelation. Gelation would not be a serious problem if cold storage of the concentrate is avoided, but, with increasing v o l u m e of milk processed coupled with shorter w o r k weeks, storage of concentrates before sterilization may be unavoidable. In this situation a better definition and a clear understanding of the changes is essential to solution of the problem,

ACKNOWLEDGMENT

The authors would like to thank Barry Vessy

1 Harwarlkar, V. R. 1982. Age gelation of sterilized milks. In Developments of dairy chemistry, Part 1: Proteins. P. F. Fox, ed. Appl. Sci. Publ. Co., London, England (in press). 2 Harwalkar, V. R., and H. J. Freeman. 1978. Effect of added phosphate and storage on changes in ultra-high temperature short-time sterilized concentrated skimmilk. Netherlands Milk Dairy J. 32:204. 3 Heintzberger, H., J. Koops, and D. Westerbeek. 1972. Gelation of sterilized canned evaporated milk. Netherlands Milk Dairy J. 26:31. 4 Law, B. A. 1979. Revision on the progress of dairy science: Enzymes of psychrotrophic bacteria and their effects on milk and milk products. J. Dairy Res. 46:573. 5 McKellar, R. C. 1981. Development of off-flavors in ultra-high temperature and pasteurized milk as a function of proteolysis. J. Dairy Sci. 64:2138. 6 Standard Methods for the Examination of Dairy Products. 1978. 14th ed. E. M. Marth, ed. Am. Publ. Health Assoc., Washington, DC 20036. p. 416. 7 TeWhaiti, I. E., and T. F. Fryer. 1977. The enumeration of bacteria in refrigerated milk. New Zealand J. Dairy Sci. Teehnol. 12:51. 8 Webb, B. H., and A. H. Johnson. 1965. Fundamentals of dairy chemistry. Avi Publ. Co., Westport, CT.

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