The use of a mixture of 60% groundnut oil and 40% hydrogenated oil in the peace time Army ration, has been found to maintain the serum lipids at a lower level and improve the physical efficiency of albino rats

Chatterjee, A. K.

Prasad, N. N.

Saraswathy, S.

English

Defence Science Journal

EFFECT OF SUBSTITUTION OF HYDROGENATED OIL BY A MIXTURE OF GROUNDNUT OIL AND HYDROGENATED OIL S. SARASWATHY $, N. N. PRASAD Defence Food Research Laboratory, Mysore AND A. K. CHATTERJEE Defence Researoh & Development Establishment, Gwalior (Received 22 Berptember 1973) The use of a mixture of 60% groundnut oil and 40% hydrogenated oil in the peace time Army ration, has been found to maintain the serum lipids at a lower level and improve the physical efficiency of albino rats. Hydrogenated oil constitutes the major source of dietary fat for the Armed Forces. Although i t  is considered suitable from the point of view of logistics, nutrition and culinary practices, i t  is very low in polyunsaturated fatty acids (PUFA). Significant correlations have been established between PUFA content of the dietary fat, hyper cholesterolaemia and atherosclero~isl~2~~~~. A high cbolesterol level in blood has been associated with coronary diseases (CHD) and it has been considered one of the major risk fac- tors for the development of CHD. The PUPA content of hydrogenated oil issued to the Armed Forcesis of the order of 2% and its bio- logical availability is limited to a great extent by losses due to auto-oxidation during storage and polymeri- . sation in the processes like deepxfat frying etc. The opt'imum level of fat or the PUPA in human diet has not yet been precisely defined . However, the American Heart Association has recommended that satu- ratedfats should not contribute more than 10% of the total calories in a dietary regimen where the caloric contribution of fat is not more than 30% of the total calories5. Judged by these standards aqain, the level of PUEA in the Army ration appears to be definitely on the lower side. Tn a large scale users' acceptability trial carried out by this laboratory6 earlier, it was : found that refined go~indnut oil fortified with Vitainin A was acceptable to Armed Force personnel, deployed under different climatic conditions, up t maximum of 60% of the total entitlement. The replacement' of 60% of hydrogenated oil by refined grou&ut oil will provide 12-13% PURA on total fat basis which has also been found suflFicient for minimising hypercholesterolaemia4. The present communication deals wit6 the effects of the above replacement of on the serum and the liver lipid patterns and physical efficiency of albino rats. E X P E R I M E N T A L  P R O C E D U R E  Experiments were carried out in three series. Tn series I, twenty four laboratory bred male albino rats, weighing 220 to 240 grams each, were divided into two equal groups 'A' and 'B' of twelve animals each and housed in separatecages. For collecting the base line data for the serum lipids 1 .0  to 1.5  ml of blood was drawn under slight anaesthesia by heart puncture in the fasting state. The serum was separated and the analysis was completed on the day of collection. The animals of group 'A' were kept on the normal Army ration and served as control for those of group 'By fed on the modified Army ration. The animals were weighed each week and the weekly intake of food was determined on dry basis (Table 1). Blood was collected in the fasting state a t  the end of the fourth, eighth, and twelfth week for the esti- mation of the serum lipids. At the end of the eleventh week, a swirnmi11g test was carried out to determine Week1 1st 2nd 3rd 4th 5th 6th 7th 8th 9th 10th 11th 12th Mean + SD Group 4 weeks 8 weeks 12 weeks ------ The difference between the means of the two groups i s  4,8 and 12 weeks is not s i e o a ~ t .  P < 0.05. ! 36; DEB. SOI. J., VOL. 24, OCTOBER 1974 the physicalefficiency. On completion of the trial period of twelve weeks, the animals were sacrificed and the samples of liver were collected and analysed. In series 11, two groups of male albino rats weighing 220 grams each, maintained on stock diet were fed ad libitum on normal Army ration and modified Army ration, supplemented with cholesterol a t  1.5 % level for a period of twelve weeks. Serum cholesterol levels were determined both a t  the commencement and at the end of the experiment. In series 111, sixteen male wcanling rats were divided into two groups of eight each. These groups were fed on normal Army ration and modified Army ration for 52 weeks. Physical efficiency in these groups was assessed a t  the end of 42nd and 52nd week. Preparation of t tion: Wheat flour, Bengal gram flour, salt, milk powder and about 80% of the v fat were mixed togethe as uniformly as possible. The vegetables cooked by boiling in waker were added to the above mixture to prepare a dough. Chapaties were prepared by baking. The onions and boiled vegeta- bles were chopped and fried in the remaining 20% of the fat; sugar, cut sweet lime and chapaties torn into small pieces, were mixed and the mixture was passed through a mincer. The material, sufficient to last for one week, was packed in paper foil laminate pouches and stored in the deep freeze. The proximate analysis is shown in Table 2. TABLE 2 COMPOSITION F THE EXPERIMENTAL RATIONS c Normal Army Modified Army Normal Army Modified Army ration (for ration (for ration (for ration (for rats of group 'A') rats of group 'B') rats of group 'A') rats of group 'B') (gm) (gm) (gm) -----_.-d---..-_)___ (gm) Wheat flour 600 600 Pot&to (fresh) 110 110 Bengal gram flour 90 90 Onion 60 c 60 Whole milk powder 44 44 Fruits (Sweet lime). 50 50 Suorose 90 90 Oil hydrogenated 70 - Salt 20 20 6 o % r  oundnut oil and 40% Vegetable fresh (Carrots) 180 180 hy rogeneted oil mixture - 70 -- - - F'roximate analysis (%): Moisture40.5, dried feed-59.5, F a t 9 . 5 ,  Protein-20.2, Carbohydrate-63.4, Ash-!. 8 and Crude fibre-5.1. Assessment of phtjsiccbl e$ciency : 0.5 ml of blood was drawn in the fasting state from each animal under slight anaesthesia by heart puncture and the fasting lactic acid level was determined according to Barker and Summerson7. The rats were kept a t  rest for one hour and then kept in the swimming tank for one hour. 0.51111 of blood was againcollected at the end of the test and the lactic acid level re-estimated. Dettrmimtion of serum lipids : The blood, after collection was allowed to clot a t  room temperature and the serum separated by centrifuging. The analysis was completed on the day of collection. The total cholesterol was estimated according to Searcy & Bergquiste ; Phospholipids according to King & Woottong; and Total esterified fatty acids (TEFA) and triglycerides according to Reinhold, Yonan & Greshmanl0. Determination.of liver lipids : The specimen of liver was weighed and dried a t  80' to 90' C to constant weight. The dried material was then powdered and kept in the deep freeze. The following method was adopted for estimation of liver lipids. The analysis was completed in a week. 0.5 to 0.8 grams of the dried liver was extracted with petroleum ether (40"-60°C) in a Soxhlet appa- ratus. After removing the solvent, the residuallipid was weighed, dissolved in chloroform and solution made upto 100 ml with the same solvent. For the estimation of total cholesterol, phospholipids and TEFA 0.5 ml of chloroform solution were separately evaporated to dryness and the methods for the serwn wqe followed. &ee cholesterol was determined by the method of Searey & Bergquists after evaporating 1 ml of the chloroform solution to dryness. The value of triglyceride was calculated according to the recommendation of the American Association of Clinical Chemists. The figures for esterifled cholesterol were obtained by subtracting the values of free cholesterol from those of the total cholesterol. The - data of various parameters were statistically analysed. The significance of the difference of the means was mlculated according to the students' 't' test (P less then 0.05). R E S U L T S  A N D  D I S C U S S I O N  It is observed from, Table 3 that the pattern of changes in the various serum lipids in the normal Army 136 ~~ARASWATHY eta1 : Substitution of Hydrogenated Oil by Mixture of Groundnut'Oil -- Group Week No. of rats TEFA Cholesterol Phospbolipid Triglyoeride C/P ratio (meqllit) (mg/lOO ml) (mg1100 ml) (mg/100 ml) Meanf SD MeanfSD MeaniSD Meanf SD Meanf D A A initial 4th 8th 12th B initial 4th 8th 12th *Only four samples of serum were analysed. t Significantly different from the preceding value. $ Significantly different from the values of group 'A'. LIVER LIPIDS OF RATS FED ON ARMY RATION AND MODIFIED ARMY RATION FOR 12 WEEKS No. of rats fed on modi6ed ration =8 No. of rats fed on Army ration =7 - - Army ration Modified Army ration - Moisture (gm/100 gm) 69.80fl-20 68.80fl.40t , Total lipids (gm/100 gm) 5.77&2.10 7.49f2.12f TEFA (meq/100 gm) 10.03&1.74 17-15f3.40 Total cholesterol (mglgm) 3.66+0.40 4.81f 0.87* l?ree cholesterol (mglgm) 2.22f0.38 2.23f 0.64t Phospholipids (gm/100 gm) 1-76kO.16 2.741 0.70* Triglycerides (gm/100 gm) 1.82&0.34 2-70f 1.55. tNot significant. "Significant. TABLE 5 SERUM OHOLESTEROL AFTER A FEEDING PERIOD O F  12 WEEKS . Group - -- - - - - - Significance of the Week No. of rats Choles@rol (mg/100 difference of the ml) Mean f SD mean < P-O.05 Army ration with 1.5% cholesterol initial 6 59.5k3.2 12th 6 132.8&13.7 Highly Significant Modified Army ration with 1.5% cholesterol initial 6 59.5 h3.2 12th 5 103.7&11.6 ration and the modified Army ration are-almost similar. The values for TEFA and triglycerides in the case of Army ration remained-at a higher level throughout the study upto 8tb week and the concentration of TEFL, triglycerides and C/P ratio in both the groups showed a significant elevation a t  the end of feed- ing for 8 weeks. At the esd of 12 weeks TEFA and triglycerides in group B showed a decline while those for group A showed a significant rise; however the means of these values for group A and B were not sig- nificantly different. No significant changes were found in cholesterol concentrations between the two groups throughout tbe study, although slight variation was found within the groups. The concentration of phospholipid in each group tended to rise a t  four weeks, and thereafter declined gradually to below the initial levels, a t  the end of 12 weeks, the differences between the groups were not significant a t  any stage. Thus the com- parison of the figures for the various serum lipids indicated that the rats of groups A and B varied in a similar manner, the notable difference being that TEFA and triglyceride levels of group B tended to remain significantly a t  lower levels as compared to the other group. No significant difference is observed in the intake of food by the two groups (Table 2) and thus the difference in the lipid levels between the two groups is due to the difference in the nature of the fat consumed. The data recorded in Table 4 indicate that after a period of 12 weeks, the livers of the rats of group 'A' kept on the normal Army ratioa show significantly lower levels of total cholesterol, TEFA, triglycerides and phospholipids than the raks of group B fed on modified Army ration. 137 ~ E F .  Sa. Y., VOL. 24 OCTOBER 1934 Series Group NO. of Before exercise After exercise rats Mean &SD Mean &SD --- -.- I A  7 13-60&4.30 37.70&8.30 B 9 13.80&3.20 23.80f 4.80* There is no significant difference in serum cholesterol levels between the rats of group A and I3 throughout the trial (Table 3). It is observed from Table 5 that on supplementation of the diet with 1.5% cholesterol, a highly significant difference exists in the serum cBolestero1 level of the two groups of rats after a period of twelve weeks, the ration containing the hydrogenated oil as the source of fat being decidedly more hypercholesterolaemic than that containing a mixture of hydrogenated oiland groundnut oil as the source of fat. , The results of the test of physical efficiency (Table 6) indicate a significantly lower level of blood lactic , acid after exercise in the rats of group 'By than in the rats of group 'A' . This trend of improved physical efficiency was observed in rats maintained on modified Army ration during 11 weeks, 42 weeks and 52 weeks. The increased concentration of PUPA in the modified Army ration seems to improve the physical effici- ency of the rats. A similar observation was recorded by Scheerl1 and co-workers with rats fed on varying quantities of cotton seed oil containing 1% linoleic acid. Thephad observed an increase in the swimming time of the rats with an increase of the linoleic acid containing cotton seed oil in the diet. C O N C L U S I O N  The use of a mixture of 60% groundnut oil and 40% hydrogenated oil in place of only hydrogenadted oil in Army ration has been found to maintain serum lipids a t  a lower level. The rats fed on the modified Army ration show a higher physical efficiency, as observed by the swim- ming test, than those fed on the normal A m y  ration. The livers of rats maintained on the modified Army ration, for a period of 12 weeks, contain a higher concentration of cholesterol, TEFA, triglycerides and phospholipids than those of rats fed on the normal Army ration for the same period. A C X N O W L E D G E M E N T S  The authors are grateful to Dr. H. Nath, Director, DB'RL, for providing overdl guidance in this in- vestigation. Thanks are also due to Dr. D. B. Parihar, Assistant Director, for his constant encouragement and helping us in the preparation of the paper. Thanks are also duc to Shri K. R, Viswanathan for his valuable assistance in analytical work. R E F E R E N C E S  1. KINSELL, L.W., MICHAELIS, G.D., PATRIDGE, T.W., B O L ~ G ,  224. L.A., BALCH, H.E., & COCIERANE, G.C., J. Olin. Nutr., 1 (1953), 2. HEGSTED, D.M., MC CANDY, R.B., MYERS, M.L. & STARE, F.J., Amer. J. Clim. Nutr., 17 (1965), 281. 3. &NOLAIR, H.M., Lancet, 270 (1956), 381. 4. JAGANNATHAN, S.N., J. Nutrition, 77 (1962), 317.$ 6. PAGB, I.H., ALLBN, J.R., CHAMBERL-, W.F., KEY, A., STBMLER, J.& STARE, F.J., Circulation 23 (1961), 133 cited in "Modern Nutrition in Health and Disease", P. 920, edited by Wohl and Goodhart, Published by Lea and Febiger, (1968). 6. KRYA, S.S., VIDYASAUAR, K. & NATH, H., !/'he Ind. J .  Nutr. Dietet., 9 (1972), 145. 7. BAREBB, S.B. & SUMMERSON, W. H., J. Biol. Chem., 138 (1941), 534. 8. SNARCY, R.L. & BERGQUIST, L.M., Clim. Chern. Acta, 51 (1960), 192. 9. &a, E.J. & WOOTTON, I.D.P., "Micro-analysis in Medical Biochemistry" (J. & A .  Churchill Ltd., London W.1), (1959), 75. 10. REINHOLD, J.G., YONAN, V.L., & G E R S H ~ N ,  E.R., "Standard Methods of Clinical Chemistry", Vol. IV, edited by Sclingson, (Academic Press Inc., New York), (1963), 85. 11. SCHEER, B.T., DORST, S., COPU, J.F. & SOULE, D., Amer. J .  Physiol., 149 (1947, 194. 

Effect of Substitution of Hydrogenated Oil by a Mixture of Groundnut Oil and Hydrogenated Oil

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Effect of Substitution of Hydrogenated Oil by a Mixture of Groundnut Oil and Hydrogenated Oil

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