Formalin as a Preservative for Analytical Purpose in Milk and Milk Products Kamal Gandhit, ‘Ashok D. Aghav, *Darshan Lal “Research Scholar, MTe “7 von of Research Karnal 182001, er, ac cht, Pinal Sota dE eb, Dil Day chen, ata Dey. Intute, Karnal 182001, Haryana ABSTRACT Formalin is the only legally permitted preservative for milk and milk products. Itis added to the milk and milk products at the level of 0.4 m! (eight drops) for 100 ml or 100 grams except for ice-cream, where 0.6 mlis used for 100 ml or 100 grams as per Food Safety and Standards Act (FSSA) (2006). It is reported to interfere with the estimation of various constituents in milk and milk products. Moreover, it has been seen that there occurs a long time lapse between the collection of the samples by Food Safety Officer (FSO) and its ‘subsequent analysis which may cause conviction of innocent traders. Thus, there is a need to look for a suitable substitute of formalin to preserve samples for such length of time so that they remain fit for analysis. INTRODUCTION Consumers expect/ demand nutritious, wholesome, pure and safe food. In recent years, consumers have placed increased emphasis on food safety. Goveriments worldwide have regulated foods with two general objectives. The first is to ensure the safety and wholesomenoss of the food supply. “The second is to prevent economic fraud or deception. These objectives encompass such concepts as safety, purity, wholesomeness and value. For this reason, the registering authority or any other officer authorised by it, may carry out periodic inspection of ‘any premisas in which manufacture or process, or business, in milk or any milk product is carried on, with a view to ‘ensuring compliance with the provisions of Food Safety and, Standards Act (2006) and Rules and Regulations (2011). While taking a sample, notice in writing is given regarding the intention of analysis to the person from whom the sample is taken. The sample is drawn in such a manner as to render the sample to be homogeneous and representative so that the analysis can furnish reliable data of the nature and content of the article. If milk or any milk product is kept, sold or stored for sale or for distribution in sealed containers having identical label deciaration, the contents of one. or ‘more of such containers as may be required to satisfy the quantity required for proper analysis, are treated to be a art of the sample Under Food Safety and Standards Act (2006), samples of food for the purpose of analysis must be taken in clean dry botties or jars or in other suitable containers which can be closed sufficiently tight to prevent leakage, evaporation or in the case of dry substance, entrance of moisture and must be carefully sealed. ‘The preservative used in the case of samples of any milk including toned, separated and skimmed milk, standardized milk, chhanna, cream, Ice-candy, dahi, khoa or khoa based and Paneer based sweets, such as Kalakand and Burfi, can be the liquid commonly known as “formalin® that is to say, a liquid containing about 40 per cent of formaldehyde in aqueous solution in the proportion of 0.1ml (two drops) for 25mi or 25g. In case of samples of ice cream and mixed ice-cream, the amount of preservative used is 0.6m! for, 100m! of 100g (FSSAI act, 2006). TABLE: Quantity of Sample of Food to be sent to the Public Analyst '/ Director for Analysis [Article of food “Approximate quantity 20 be supplied lk 00m ‘Storied MIR/OHT Ml 250m MaaiiDani 200g Yoghurt Swestened Dani 3009) [chnanal Paneer! KnoyaSivikhand 2609) (Gheeseicheese spread 2009) [Evaporated Mik/ Condensed Milk 2009 ioe Crearn’SoftyiKutvice Candy Ice Laly 3009 alk powder/Skimmed Mik Powder 250g fant Food) Weaning Food. 5009 [Mail Food Malied Mik Food ‘300g Butter ButorOWGhoa/Margarine” "2006 |Croarvt 2509) ‘Whenever any preservative is added to a sample, the nature and quantity of the preservative added Is to be clearly noted on the label to be affixed to the container. The quantity Cf sample of food to be sent to the public analyst / Director, for analysis shall be as per specifications as shown in Table 1: So, in general it can be said that the food samples collected by the food inspectors for analysis are required to be kept for a long time. The time varies from 3 10 6 months ‘and sometimes longer. The food samples, especially some prepared foods when it reaches the laboratory for analysis, are found to be decomposed by bacteria or mold. Such samples are declared unfit for analysis and rejected. So, for the proper maintenance of the perishable food samples, use of a suitable preservative is @ must. 44 BEVERAGE & FOOD WORLD - Vol. 41 - No. 6 - JUNE 2014‘preservative’ for milk may be defined as any chemical ‘compound and/ of process which, when applied to milk, retards alterations caused by growth of microorganisms or ‘enables the physical properties, chemical composition and original nutritional value to remain unaffected by microbial spoilage. ‘The ideal moans of preserving milk samples for several days prior to compositional analysis is refrigeration as near the freezing point as possible without freezing the milk. This prevents growth of bacteria and all the chemical and physical ‘changes following such growth. Ice crystal formation usually Jeads to disruption of physical structures in the milk, making itdifficultto withdraw representative subsamples (aliquots). ‘The minimum requirement for a suitable chemical milk ‘sample preservative is that it must assure “testability of ‘sample’. This means that the milk sample maintains its ‘original composition from the time of milking to the time of analysis and that the substance does not affect the outcome of the test procedure. Other requirements are 1. Broad-spectrum Activity A critical necessity is the pernicious ability of the preservative to act against microorganisms in milk. The broader the spectrum, the better the utility. It should be cffective against all ypes of microorganisms occurring in milk. 2. Efficient Minimum Inhibitory Levels The preservative should be effective at low concentrations in milk to minimize sample dilution, minimize costs, and expedite handling procedures. High Water Solubility Because the average milk sample is about 87% water, itis imperative that the preservative have the capacity to function where the microorganisms are, i.e. in the ‘aqueous phase; high water solubility would also assure easy solution without excessive stirring, 4. Stability The preservative must be stable under most storage conditions. 5. Presence of Color Unlike cosmetic preservatives, a desirable quality of a milk preservative is that it imparts some color to the treated milk sample for indicator and safety purposes. 6. Compatibility A.good milk preservative should be just as effective in ‘pooled milk samples as in fresh samples from individual cows or other mammals and be suitable for high-fat as well as low-fat milk samples. 7. Shett-life Activity ‘An adequate milk preservative should be effective for a reasonably long period. 8. Toxicity and Disposability ‘The preservative should be non-allergenic and exhibit no demonstrable toxicity toward handlers or others coming in contact with it. Despite its necessary biocidal property it should not become an environmental hazard after disposal. 9. Economy ‘Cost should be minimal and the preservative should be readily available, 10, Dispensing Ability Dispensing in solid form, as in tablets, would be preferable to the liquid form, because of the inherent difficulty in handling liquids and the greater accuracy in dispensing solids. Formaldehyde (HCHO) Formaldehyde is commercially available as 37-40% ‘aqueous solution known as formalin. On storage, HCHO hhas the tendency to form polymers- paraformaldehyde (CH,O),, Mostly trimers known as meta-formaldehyde or tr-oxymethylene (CH,0), are formed. These trimers show ‘no reducing properties, since no free ~CHO is available. In ‘order to inhibit the polymerization, 10-15% methanol is ‘added to the commercially available formalin. Formalin being ‘a storage antiseptic and disinfectant, it is used in the preservation of various anatomical species and also food stuffs. Being carcinogenic in nature, its use for edible purposes has been prohibited, however, it has been permitted under FSSAI rules, 2011 for the preservation of milk and milk products meant for chemical analysis at a level of 0.1mi/ 25ml i.e. 0.4% (viv, vw) of milk or milk products. It acts through alkylation of amino-, carboxyl- or hydroxyl group, and probably damages nucieic acids. It kils all microorganisms, including spores. Use of formalin at such level has shown contradictory results regarding the preservabilty of samples, which could be due to various reasons, such as use of substandard formalin, storage period, storage temperature, nature of the sample preserved, etc. The preservative has also been shown to affect various physico-chemical properties as well as the fat, protein, lactose and total solids content during storage. This has created a problem and brought industry indirect confict wth the regulatory agencies. Many research workers have studied the compositional changes in milk ‘samples as affected by formalin preservation (Boghra and Borkhatriya, 2003). Effect of Formalin on Various Physico-chemical Properties of Milk 1) Effect on Freezing Point Venkateshwara Rao et al., (1950) studied the Freezing point depression in mikk samples preserved with different preservative, using Hortvet Cryoscope. Formalin at the level of 1.0% level was observed to increase the freezing Point depression of milk considerably. The freezing point depression further increased on storage. Formalin has found to be most effective in increasing the freezing point depression as compared with other preservatives, such as potassium dichromate, mercuric chloride etc. ‘The effect of formalin on FP of milk and suggested that the milk sample can be preserved with a known amount ‘of formalin, but a correction factor has to be applied to find the FP of original milk. (Girous, 1963). 2) Effect on Acidity A slight decrease in acidity of milk, on addition of Formaldehyde was reported by earlier workers. However, further research shown that the addition of Formalin caused a slight increase in the intial acidity of milk (Venkateshwara Rao etal, 1950; Armandola, 1967; Sandhu ef a, 1984). An increase in acidity of milk on immediate addition of Formalin and subsequent storage has been recorded by most workers. The increase in the acidity was also dependent on concentration of Formalin used. On the storage, acidity increased further in the same order for four- five months, thereafter the 48 BEVERAGE & FOOD WORLD - Vol. 41 - No. 6 - JUNE 20143) 4 order of increase in the acidity reversed i.e. increase in the acidity was more in the of sample containing low level of Formalin as compared to sample having higher levels of formalin. Increase in the acidity on addition of formalin is because of liberation of hydrogen ions. 7 'NHOH2OH NHS: ” / CH + HCHO = RH \ \ cooH bai Ens SRS Snes nesu enn eneeeases eens snneneeeee Mik samples preserved with 0.4% formalin were of acceptable quality when stored at 30°C for 3 months, despite an Increase an in titratable acidity, The increase in the acidity was attributed to the higher proteolytic activity in the samplo with the lower level of formalin Karmakar and ghatak (1997) noticed an immediate increase in the acidity on addition of 0.1, 0.2 and 0.4% formalin in cow milk and also on subsequent storage at refrigeration temperature for 30 days. The acidity increased from 0.126% to 0.130% with 0.4 % formalin and continued to increase to 0.185% after 90 days. Effect on Specific Gravity During the preservation of milk with formalin (one part ‘of formalin per 1500 parts of milk) for seven days, Nelson et al., (1958) observed an increase of 0.03% in SNF. ‘Armandola (1967) reported that a few drops of form: containing approximately 4% of para- formaldehyde to milk was suitable for density determination upto 4 months. Armandola (1969) again observed no change in density of milk upto two months on addition of 0.045 % of formaldehyde. Other workers also observed significant change in SNF content of milk preserved with various concentration of formalin. Similar observations were recorded by Ramaiah and Narayanan (1976) in ice cream samples when preserved with formalin. Effect on Viscosity ‘An increase in viscosity was observed in all the milk sample preserved with different level of formalin. After 2 months of storage, the buffalo milk samples become 80 viscous that it was not possible to determine their viscosity by Ostwald viscometer (Bansal and Singhal 1991). Bajaj and Rai (1993) showed an increase i viscosity of buffalo milk preserved with 0.4% formalin after 21 days at 30°C Effect on Cream Layer Formation During the storage of the milk samples at 37°C in dark, cream plug appeared within 24 hr. A white sedimentation probably due fo casein, globulin and albumin appeared at the bottom after one month. It continued to increase in the volume with the passage of time. At the end of ‘one year there were three distinct layers in the samples. ‘These were :- 1) top layer of cream plug, 2) middle layer of liquid, and 3) bottom layer of semisolid material. ‘On blending, these layers mixed radially and the resultant sample was stable. The blended sample were warmed to 40°C on water bath, and cooled to ambient temperature before analysis. Bansal and singhal (1991) while preserving the composite sample of cow and butfalo milks with 0.2, 0.3, 0.4 and 0.5% formalin (viv) and storing in glass boitles at room temperature (16- 30°C) up to one year, showed that milk separated into 46 BEVERAGE & FOOD WORLD - Vol. 41 - No. 6 - JUNE 2014 three layers. These layers were more distinct in case of buffalo milk samples as compared to cow milk samples, ‘The homogenization prior to analysis was required after 4.5 months of storage of buffalo milk for proper sampling Effect on Clot on Boiling (COB) Test Preserving the fresh cow milk by formalin 0.4% at a temperature of 30°C showed the negative COB test after 3 and 6 months. Others also reported that cow and buffalo milk samples gave positive COB after 7 and 4 months with 0.2% and 9 and 7 months with 0.3% formalin respectively. Only buffalo milk gives positive COB test (after 6 and 7 months respectively) with 0.4 ‘and 0.5 % formalin. A positive COB test with preserved utfalo milk stored at 30°C after one month was observed by Bajaj and Rai (1993). Change in Appearance Fresh cow mik samples preserved with formalin did not show any visible change in color and physical eppeaano for entra pared! storage or 3 mons IC however efter 45 days of storage, allmik samples preserved with Hach t 0.4% level developed black color at the bottom and along the sides of the sample bottle. Bajaj and Rai (1993) did not find apparent change in the color of cow and buffalo milk samples preserved with 0.4% of formalin for 6 months at 30°C, however, buffalo milk show the signs of the curdling at the retrigeration temperature Effect of Formalin on Milk Constituents 1) Effect on SNF/TS Proservation of milk with varying level of formalin for different time periods at room and refrigeration temperature has been found io have no effect on SNF/ TS content of milk as estimated gravimetrically. However, some discrepancies were reported by some workers when SNF was calculated using lactometer readings. Effect on Milk Fat a) Gerber Method Formalin gave the problem during the fat estimation by he Gerber metho, and giv the wort values The fat contents of formaldehyde treated milk samples could not be determined accurately because curd is nat dissolved in the heat generated by 81 % H,SO,, hence certain modifications in the Gerber method such as repeated heating of milk in the butyrometer at 65°C and Tepeated entrifugations, shaking, were suggested, which however, give enormously high values. Ithas been reported by the authors that fat values decreased with increase of hydrometer reading, formalin ‘concentration and also the period of storage. Karmakar and Ghatak (1995,1997) have reported rno change in Gerber fat values in milk samples preserved with 0.4 and 0.6% formalin and stored for varying period to 1 year at room temperature. Rose Gottlieb/ Mojonnier Method “There was no immediate impact of formalin addition on Mojonnier fat test. Narayanan (1973) and Sandhu etal, (1984) reported no change in the fat value determined by Rose Gottieb method of the ‘milk samples preserved with 0.4 and 0.6% formalin. for 6 months to 1 year, Des raj and Singhal (1989), however reported that Rose Gottlieb method 8) n b)exhibited low fat values in milk samples preserved ‘with 0.4% formalin, though the effect of formalin on rose Gottlieb method was less and inconsistent as compared to Gerber method. The fevel of formalin did not significantly affect the values determined by the Rose- Gottlieb method, although a steady decrease was observed on the storage of the preserved milk sample. On an average, the fat value decreased by about 20% after one storage in all the samples ‘The decrease was duo to the hardening effect of formalin on protein that entangles some of the fat which is difficult to recover on the treatment with ammonia. ¢) Instrumental Method ‘There are the differences of opinion on the effect of preservatives on the results of milko -tester. Several workers reported that the formaldehyde did not alter the results of the fat estimation by milko-tester. However reports of some workers indicated the variability in the milko tester fat tests due to the presence of formalin as a milk preservative. Low fat values by milko-tester as compared to Gerber fat values have ben reported by Marquering (1971) {or the milk sample preserved with formalin. Milko- tester has been reported to be unsatistactory for the determination ofthe fat in the formalin preserved milk samples. The preservation of the milk sample with formalin for 3 days at ambient temperature (20- 30 °C) did not distort the subsequent Milko tester value whereas, an average reduction of 0.03% fat was observed after 3 days and further storage for longer periods gave unreliable resutt. Formalin as a preservative has been reported as unsuitable for fat determination in milk using mini IRMA. 3) Effect on Proteins Itis established fact that the formaldehyde reacts with amino group of protein to form a harder formal-protein complex. Hence, it was desired to compare the various protein estimation methods for their efficiency and results. The methods used for the detection of protein are formal titration, kjeldah! method and dye binding method. The addition of formaldehyde to milk depressed the formation of soluble casein. It was observed that no soluble casein was found on addition of 20 mM formaldehyde. The reduced solubility was due to the introduction of cross links by formaldehyde along casein components. It is estimated fact that formaldehyde reacts with amino groups of protein to form a harder formal-protein complex. Bector and Narayanan (1973) and Bansal (1989) reported no change in protein either ‘on addition of formalin or during storage up to six months with kjeldah! method, the variations were within the ‘experimental error limits but Jandal (1985) observed a ‘decrease in protein on addition of 0.4% formalin, ‘Addition of formalin caused an immediate decrease inprotein content of cow and biffalo milk by Dye-binding method. These values further kept on dectining upto ‘one month and thereafter became more or less constant in both types of milk. The differences between per cent decrease in protein of cow and buffalo milk samples Is due difference in Dye-Binding Capacity (DBC) of different fractions of protein, ‘The decrease in protein values is due to blocking of active sites of protein by formalin, which is responsible {or forming the protein-dye complex. A steady state after one month of storage was reached because the available active sites got fully saturated with formaldehyde and hence-equilibrium was achieved (Bajaj and Rai, 1993). Bajaj and Rai (1993) reported that tormaidehyde adversely affects the protein content by Lowery’s method. The values further decreases on storage and became constant after one month of storage. The decrease in protein was due to reaction of formaldehyde with tyrosine. 4) Effect on Lactose Formaldehyde is known to have effect on fat and protein estimation. However, lactose content didn’t change, either on addition of formalin or during subsequent storage upto six months, because of the fact that lactose did not play any role towards increase in acidity in formalin treated milk, The increase in acidity of milk is merely due to formalin-amino-reactions. Sandhu et al. (1984) and Bansal (1989) reported no change in lactose by Lane-Eynon method in formalin preserved samples during storage upto one year. ‘Addition of formalin did not influence the lactose determination initially on zero day as well as during ‘subsequent storage (Grimbleby, 1956 and Bajaj and Rai, 1993). The lactose contents by Picrio acid method on zero day did not change but on subsequent storage, initially there was a decrease followed by erratic results {or lactose content. The possible reasons may be (1) the incomplete precipitation of protein (2) he absorption of lactose by the coagulated protein and (3) extensive hydration of protein coagulum which resulted in ‘concentration of lactose in the serum (Bajaj and Rai, 1993).Microbial analysis of the preserved sample showed that formalin inhibited the growth of microorganisms responsible for the degradation of lactose. Preservation of Milk Products with Formalin Mukherjee and Mathew (1974) found that spraying of formaldehyde at the rate of 0.4% on cheese, channa and khoa allowed the sample tobe stored satisfactorily for more than 10 months. Dinkarand Sharma (1989) reported that a dose of 0.2 mlof formalin per 25 g of khoa was found to be most effective in controling the development of lactic acidity, volatile acidity and water soluble protein. No significant effect ‘was observed by them on moisture, fat and total protein content, however vitamin A and vitamin C level decreased. Paneer could be preserved well in sterilized and stoppered bottle for 6 days with the addition of formalin (0.4% viw) However on 21 days of storage, the samples becomes moldy and there was decrease in fat and total nitrogen content and increase in water soluble nitrogen, tyrosine and acidity ‘of sample. Addition of formalin at the rate of 0.4% can preserve the sample of plain ice-cream (at 25-30c) for 2 months. Further storage decrease the total solid, fat and ‘sucrose content appreciably. Therefore it was recommended that ice cream sample preserved with 0.4% formalin should be analyzed within a period of 2 months to check its ‘compositional standards. CONCLUSION Formalin interferes with the estimation of the various constituents whose minimum or maximum limits are fixed ‘Contd. on Page 49 47 BEVERAGE & FOOD WORLD - Vol. 41 - No. 6 - JUNE 2014considerably lower than in tomato ketchup. Thick sauces made from fruits and vegetables other than tomatoes are rot called ketchup (Verma and Joshi, 2000). Tomato ketchup is covered by USDA Standards for Grades and by an FDA Standard of Identity. Only a product made from whole tomatoes may be called ketchup without qualification. The pulp quality determines the final product quality and in inferior grades of ketchup made from poor pulp, the flavor of the ulp must be overcome by added spices. The grade is based ‘on color, consistency, absence of defects, and flavor. In this case consistency is defined as the viscosity of the product, and the tendency to hold its liquid portion in suspension. The grade A and B are defined as having good consistency, while Grade C should be of fairly good consistency (Diane et al, 1998). ‘Tomato ketchup is a'heterogeneous, spiced product, processed basically from either cold or hot extracted tomato or directly from concentrates, purees and tomato paste, Viscosity is an important attribute from the engineering and consumer view points. Tomato ketchup obtains its viscosity from naturally occurring pectic substanoes in fruits. Tomato with less pectin may result in reduced consistency, and other factors such as enzymatic degradations, pectin/protein interaction, pulp content, homogenization process and concentration may also affect the consistency of tomato products (Hilal and Feramuz, 2004). The tamarind tree is a native of Africa and Tropical Asia land is growin for its pulpy frult tamarind (Tamarindious indica), which has a sweet sour taste. The tamarind grows widely in tropical and subtropical regions of the world, The tamarind tree can produce an annual fruit yield ranging from 150-500 kg/tree. The tamarind fruits are mainly produced in India and Thailand where it is grown on a commercial scale and exported both in fresh and processed form (Gebauer et ai, 2003). The most valuable and commonly used part of the tamarind tree is the fruit. The pulp constitutes 30-50% of the ripe fruit, the shell and fibre account for 11-30% and the seed about 25-40%, Tamarind pulp is used as a raw material for the manufacture of several industrial ssuch as tamarind juice concentrate (TJC), tamarind pulp powder (TPP), tartaric acid, pectin, tartarates and alcohol. The pulp is also used in traditional medicine, ‘Tamarind Kernel Powder produced from seeds is another commercial product and is often reported upon in commercial digests (EI-Siddig et al,, 2004). The detailed physical and engineering properties of tamarind pulp and seeds are done by Bhattacharya et al, (1993). In India it is the chief acidifying agent in curries, chutneys and sauces. The tamarind sauce is prepated from tamarind pulp by adding sugar, salt, spices, etc and then heating the sauce to the desired consistency. ‘The hydrocolloids such as guar gum, CMC and sodium, alginate can be added to increase the viscosity of the sauce at lower total solids content. The consistency/viscosity of ketchup is an imporiant attribute from the engineering and ‘consumer viewpoint (Rani and Bains, 1987). Knowledge of the theological properties of fluid and semisolid foodstuffs is Important in the design of flow processes in quality control, in storage and processing stability measurements and in understanding and designing texture, Therefore, reliable and accurate rheological data are necessary for designing and optimization of various unit operations (pumping, mixing, heating etc) and ensuring product acoeptabilty since the products with improper consistency may be graded as unacceptable, or sold at a lower price. Sears (90 I pets, cmos ryan 6). Fit pete pocuc,Tata typ. Det, campeon eee rerisines aces emamenis Tiana avait nes) sn ua ood ngroweg 1-70 eens eaten asd EW itl ti! Seeker of eng tne Ses en Sep i VE Tonalin soso soon of and PEG cg goton eo (ie Br ay 3 tt PY acu lh Sac rege omen cnc ‘ots Yool Tasogy Bate Bea SSOP et tart nd): tay tien sd Soles tira totaly ese Sd 3 Tp oon mtn pe a nt ‘Hilal, S. and Fevamuz, 1 (209, Eta of sae Pca n he shealogcal rperin of ‘nema nsec ay ears soe fel m0) torent meaner ena 202 Baa ear 1s (a Sec pn is ay of gon es Bere ih el iat crmtreatees SH a Peseta Sonne erase Keine ota Pics 79 Oster got i Powter ned Tr Suen 3ol, ed Meng, goes Om pt fara ond ola Bling nation Po rma aia Pn an nag Siebrand ober nb etn aay Rew ah hp 7 ieee Fata one! Contd. from Page 47 Formalin as a Preservative for Analytical Purpose in Milk and MIlk Products under the national mandatory and voluntary standards. ‘Amongst the various chemical preservatives formalin has been assessed as the most effective preservative for milk and milk product samples at ambient temperature. However the efficiency of formafin in maintaining the compositional profile of milk and milk products intactis questionable based €n the wide scientific research support. Quality of formalin Used wil be of great concern in the present context of the quality awareness among the manufacturers and the ‘consumers with global competition and stringent food laws. The use of formalin has shown contradictory results formalin, storage period, storage temperature, nature of the sample preserved etc. ERENCES EARS fy rant spain te 8 ‘ari Er Pwo ot ks ra et 128 (Ce Dey Si ma] ic art. (82 Conprate Er tvs nates nad yard et re ore eigen cae Pans SS LEY Fea ht pao cone ooo pon ad wee "X ard Srant @P/e8) enero of sare Wt Fema exten acid sos ere Sh He i rai KM 87sec organ ok cs "ed oR eden eth Ey SEF at & 8 stokes yy enh tof rg tgs Foe ‘nator ng Dut gr Dharnaran C8190) Etec esr ore 49 BEVERAGE & FOOD WORLD - Vol. 41 - No. 6 - JUNE 2014