India. J. A.im. S,i. O (lOJ: 918-24, (klober 1973

Studies on the cholesterol content of cow and buffalo ghee

M. P. BINDAL and M. K. JAINI

.National Dairy Research Institute, Kamal, Haryana

Received: 2 January 1973

ABSTRACT

A study WM conducted on 320 la.ctating cows a.nd liD la.ctating bu.ffalOC3 to find out the cholesterol ccllteat of co.,. .. - and buffalo ghee. The total cholesterol c::untents were significantly higher in cow ghee (330 mg~~) than in buffalo ghee (275 mg%.). Variafions due to season \\,cre· highly significant in both types pf ghcc. Tot.:d cholf'".Steroi at the unset of lactalion was ralber high (554 mg% for cow, and 51 6 mg% for bulf.loes). T wo peaks, one al the mid­lacta tiorl stage a.nd the other wwards the end of the lactation., were noticed. Signifir.a.ntly diff~rcut level<; of total cho[e5tecol were observed in dt!ji ghee (292 mgll/~), direct cream (DC) ghce (303 mg%J and but ter-fat (318 mg%) m"de from cow milk. D<s' (263% ) and DC ghee (275 mg%,) prepared from buffalo milk showed significant diffc..:rcn,ces in tota l cholc$lcrol content when cOInpared with h'..1tter-fat (294 mg%L but diIfert:nc'~ uetween them were not significant. Total cholc~tt:rul cOOten," of butter-fat pj·epar C",d ftom m:.s,yjtic milk w,,~ very high (129 mg% for cows and 625 mg?.-~ for buffa.loes) . Significantly higher lcvel$ of cholesterol were ob~erved in gheepn:parcd from fore-milk (967 mg~·:; for cows and297 mg~/r:I fur buffaloes) th.an in ghcc prepared f!'Om strippings (324 mg% for' cows and 262 mg% for bum'l.10t~). Clarifica.tion temperature had no effect on the cholester-ol C(lntcIlt of ghee.

Free choles.terol conlcI~t wa~ significant.ly higher in cow ghee (283 mg~·~) than in buffalo gh~c (212 mg%), wheJ"eas ,ha t of e~teriflcd cholCbterol was; significantty higher in buffalo ghee. VariationH due to season were highly :l.ignificant in ea.terified cholesterol but not jn free ch<Jle~teroL Colo~trill fat differed markedly from normal nlitk f.at (ghcc) in regard to its free and t::!!itcrifieu choles terol content.

The present paper records variations in the free, esterified and total chole~terol contents of ghce with factors such as species, stage of lactation, disease, season , etc.

MATERIALS AND !vIETHODS

Feeding and ma1lagemenl of animals Three hundred and twenty lactating

cows (96 each of Sahiwal and Tharparkar, 48 Red Sindhi and 80 Brown Swiss) and 60 Murrah laetaling buffaloes were utilized. Calving was spread throughout the year. All the animals were managed and housed under identical conditions in open-type barns.

The animals were fed balanced rat ions consisting of green roughage, dry fodder and concentrate mixture (groundnut-cake, 35%; wheat bran, 25%; barley, 38%; common salt, 1%; and mineral mixture, .

Presen t address: [Chief Chem.i:i.[ , PL.4BO, Djvision of Da.iry Chemistry.

I %J. All animals were given 0·5 kg concentrate as let-down ralion. Animals yielding more than 4 kg/day additionally received I kg concentrate for every extra 2·5 and 2 kg milk produced by cows and buffaloes respectively. The animals were provided with 2/3rd leguminous and 1/3rd non-leguminous roughage during wintcr (December to February), 1/2 I"gu­minou. and 1/2 non-leguminous roughag.-: during spring (~larch to May), and all non-leguminous roughage during summer and autumn (.July to November). Non­leguminous fodders consisted of maize, oat and sorghum (green or in silage form), and leguminous fodders of berseem, lucerne and cowpea (green or in silage form). The animals had free access to drinking water.

Milk F or variations in total cholesterol

content due to species and season, 48 composite samples each were collected at

916

October 1973] CHOLESTEROL CONTE:>IT OF COW AND BUFFALO CREE

weekly intervals from cows and buffaloes. F or variations in free and esterified cholesterol contents, 24 sample. each of cow and buffalo hcrds were collected at fortnightly intervals throughout the year.

For variations due to stage of lacta­tion, samples were collected daily, imme­dia tely after parturition, from 4 Sahiwal cows and 4 buffaloes which had calved on the same: day. Milk from all the milkings of I day was pooled together and composite samples were drawn. At the end of 9 days, milk samples were collccted at fortnightly intervals for the remaining lactation period. .

For seasonal variations, samples of herd milk were collected fortnigh tly from separately pooled milk of cows and buffaloes maintained at the farm of the Imtitute .. The whole year was divided into winter, spring, summer and autumn.

For studying the effect of method of isolation of milk-fat, 10 composile samples each of cow and buffalo milk were con­verted into ghee/bu!ter-fat.

For st,;dying the effect of mastitis, 12 composIte samples eaeh were collected from the milk of cows and buffaloes suffering from clinical mastitis.

For variations due. to clarification temperature, 15 herd milk samples each of cow and buffalo were converted into ghce, using 110" and 145"C as the temperature of clarification.

For the cholesterol contents offorc-milk and stripping" 12 compo,i!e samples each of fore-milk and strippings Were collccl­ed separately from cows and buffaloes.

Preparation if ghcelbutter-fat Desi ghec from normal milk and

direct cream (DC) ghee from colostrum were prepared essentially according to the method described bv Srinivasan and Anantakrishnan (1964). Temperature used to clarify the butler/cream was lLOoC. Butler-fat was isolated from milk by the lSI method (lSI, 1966).

AnalYsis if ghee/ butter-fat Free and esterified cholesterol were

estimated by the TLC-cum-colorimetric method reported from this laboratory (Binda! and Jain, 1972). Total cholesterol

was estimated according to the procedure of Bindal and Jain (1973).

RESULTS AND DISCUSSION

Variations in total cholesterol COli tent of gkt Due to species and season." Total choles­

terol content (Fig. I) of cow and buffalo ghcc varied between 302 and 361 mg% (average value, 330 nfg%), and between 202 and 312 mg% (average value, 275 mg%) re~pectively. The,e variations were highly significant (P<O· 01). Homer and Virtanen (1967) observed that milk­fat contains 300 to 345 mg% of total cholesterol, values .imilar to those found in this study. However, wide variations in total cholesterol content of milk-fat of cow were reported by Laskowski (J 962; fide Homer and Virtanen, 1967). Information on the cholesterol contents of Indian cow and buffalo ghee is very limited. Gulvady.t al. (l952) reported rather low values for the total cholesterol content of both cow (I to 6 mg/JOO ml milk) and buffalo milk (I to 8 mg/IOO ml milk), while Laxminarayana and Dastur (1968) obtained a significantly higher cholesterol content in cow milk (I5· 8 mgt LOO g milk) than in buffalo milk (4·8 mgt 100 g milk).

;l= ~

~ E .!i " ru

"0 L <.)

The variations (Table 5, Fig. I) in 480

440

420

400

380

-360

340

3~

JOO

~SO

260

25 2

lOT D J F

M!'1T'!tr.o;

Fi.g. 1. Scas.unalts.pecies variat.ion in total cho­le;;h::'L'ul content in ghe~. 1~ Cow ghr:e; 2. buffalo ghec,

919

I1lNDAL AND JAIN [Vol. 43, No. 10

500

480 ... 4~O

440

420

400

380

; 360

'" !, 340 "0 !i 320 m

" '5 300

"'" 0 250

260

250 HiT , , .

1 2 3 4 ~ G 7 8 9 10

' lp.ctiJ.tiol"l (d a}'s}

Fig. "). VariationB in total choles.terol content of colostral fa t. 1. COWj 2) bllfFalo.

total cholesterol content of ghee due to season were highly significant (P<O· 0 1) . Thc cholesterol contt:nt in co", ghee during winter and spring was significantly higher than during summer and autumn, but the differences during winter and spring and during summer a nd autumn were not significan t. The cholesterol content in buffalo ghec during spring wa s significantly higher than tbat during winter, ~ummer and autumn. However, the differences among the average choles­terol content during wioter, SUllmer and autumn were not significant. A higher cholesterol content occurred during winter and spl'iog when the anim als were offered mostly leguminous feed.

These fe, ults are similar 10 those reported by Gulvady tl at, ( 1952) , who noticed peak pcriods in February and in August, the value, in August being sligh tly )"ss. Azim and Mustansir ( 1971) reported higher values during winter and

920

(60

440

420

400

380

360

~J"O = E - 320 '2 "* 300 ~

,g 280 U

260

240

220

210

lOT , I 2 3 4 5 6 7 8 10

L<l Clalion (mcri"tl'1s)

Fig. 3. Variation! in total cholesterol content of ghee. 1, Cow; 2, buffalo.

autumn, and comparatively lower values during rainy and spring seasons in Paki..tan. The lower cholesterol values could be due to th e intake of more water and green fodder by the animals during these periods with consequent p roduction of low-fat mi lk.

Due to ,tage if laclation.' This has been considered under two su bheads, viz. colOSlcal period and post.colostral pet>lod.

C OLOSTRAL ~ERIOD; The colostral fat in cow milk contained a higher level of cholesterol than that in buffalo. Choles· terol con tent of co lostral fat, both i.n cow and buffalo, kept falling with the progress of lactation (Fig. 2). The highest chole,;teml content was obtained on the fint day after p ari urition, followed by a gradual fa ll on the following days. On the 6th day after calving, when the colostrum ceased to clot 00 boiling, total cholesterol level in cow and buffalo ghee

October 1973J CHOLESTEROL CO:-lTENT OF COW AND BUFFALO OREE

Table L Total cholesterol content ofghee/butter.fat (in rng%)

Ghccjbutter-fat from cow milk' Ghec/buttcr-fat from buff~lo milk"

Deri ghee DC gheo Butter-fat De.ri ghee DC ghee Butta-fat

200 292 306 261- 284 295

297 3J2 320 277 281 297

2% 312 321 2J7 272 291

400 313 32;; 268 277 29B

291- 306 321- 267 275 293

289 302 320 261- 272 295

290 307 315 2GB 281 290

300 316 325 265 281 301

291 305 3J8 256 274 30J

287 300 311 252 262 283

Aver'lge 293'4 306·7 SIB·7 263·6 275· 9 294'4

", Critical difference, 1· sa i """, criti<=al differt.:nce. 3·94.

fell down to 336 and 265 mg%, respectively. At the end of 9 days, these values were reduced further to 283 mg%, in cow and 256 mg% in buffalo. The results ob­tained are in accordance with the earlier reports on the total cholesterol content of milk or milk-fat of exotic cows. Homer and Virtanen (l967) reported peak cholesterol valu~s in the first milkings with a decreasing trend in subsequent milkings. Steger (1962) also reported " high cholesterol content during the colostral period.

POST-CO LOSTRAL PERIOD : Cholesterol content was minimum within I month after parturition, both in cow and buffalo ghee. followed by a gradually increasin g trend till the 9th month after calving (Fig. 3) . When the animals almost dried off in the 10th mon th, there was a sudden rise ill the cholesterol content, the r isc being steep in cow ghee than in buffalo ghcc. Steger (1962) , and Hom"'f and Virtancn (1967) rcported relatively higher

. cholesterol levels in milk-fat at mid­lactation stage and a sudden rise in cholesterol content towards the end of the lactation. Gulvady _t at. ( 1952) reported a similar trend in cholesterol levels of cow

and buffalo milk, although their values were lower than the present data.

Due to method of isolating milk-fat: Desi ghee, direc t cream (DC) ghee and butter­rat were prepared from the same lot of milk and analysed for their cholesterol contents (Table I ) . The variations in total cholesterol content in the 3 types of milk-fat wcre highly significant (P<;O'OI) both [or the cow and the b uffalo. In milk-fat of cow, the 3 types showed significantly different levels of choles terol. The average incr",,~c from ciesi to DC ghce was 4·fl7% and from DC ghcc to buW' r-fat 3··91 '.Yo. The overall increase from desi ghee to lmtter-fat amounted to 8 ·132%. In milk-fat of buffalo, both dosi and DC ghee showed significant differences in cholesterol content compared with butter-fat, but no significant difference was noticeable between desi and DC ghee thcm~e1vcs. The average increase from dui to DC ghee was 4'69 %, and that from desi ghcc to butter-fat was 1l·6%. TIle increase from DC ghec to butter-fat was 6·7%,. Appreciable amounts of milk cholesterol are a ssociated with proteins (Kayser and Tachkova. 1958; Mulder and Zuidhof, 1958). This protein-bound

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BINDAL AND JAIN [Vol. 43, No. 10

Table 2. Total cholesttrol content of butter-fat prepared from milk of cow. and buffaloe,

suffering frow clinical mastitl!'t

Total cholesterol level (ms%)

Cow Buffalo

512 437

510 370

762 1,176

756 217

611 225

572 974

562 773

599 1,001

847 624

%0 469

1,009 866

.,050 375

Aveca.ge 729·2 625· 6 .. ---_ ... . - -- --_ . ---

't', value, 0 -094:' NS; NS, non-significan t.

cholesterol is possibly lost during separa­tion of cr~am from milk or during churning of daM. Butter-milk contains a fairly high level (503-627 rugj 100 g fat) of cholesterol. Thus cholesterol losses in skim-milk or butter-milk might possibly ""plain the lower cholesterol levels of DC and des; ghee.

Due to mastitis.. Cholesterol contcnt of butter-fat of cow and buffalo prepared from mastitic milk is given in Table 2. Variations due to specics were not statisti­cally significant, but variations due to samples in both the specics were very wide. Evidently, a disease like mastitis increased the total cholesterol level of m ilk-fat tremendously. There is little information in literature in regard to the eholcst~rol content of milk/milk-fat of animals suffering from mastitis.

Due to clarification temperature: Varia­tions in total cholesterol contents of ghee prepared at different clarification tempera­tures were not etafi.tically ~ignificant

922

('t' value, 0·7926) , both in cow and buffalo ghee (Table 3).

Chous!ero/ contmt of fore-milk and strippings Ghee prepared from fore-milk contained

significantly higher level of cholesterol than that prepared from strippings (Table 4). These variations were highly significant (PSO'OI), both in cows ('t' _value, 3·9943"') and buffaloes ('t' value, 3 ·9481 U ). Fore-milk with a lower fat level and smaller fa t globnles (Jenness and Patton, 1959) has a higher cholesterol content than strippings with a higher fat level and bigger fat globules. Cholesterol content could perhaps vary with size of

Ta.ble 3. Varia.tions in total cholesterol content of ghec prepared at different clarification

tempera.tures

T ota! cholesterol con ten t (ml5%)

Buffalo gh~e"

110"C 145"C II000e 145"C

334 333 270 268

334- 335 267 265

331 324 271 268

354- 353 280 279

351 346 294 m 350 349 281 279

337 338 277 274

356 354 281 280

330 326 275 273

330 324 216 272

333 333 284 284

350 349 283 281

330 327 270 267

332 3S1 302 100

330 326 271 268

Average 338·8 336·7 276·6 278 ·8

-- --- - --- --- - - --_ ._--oj< ~ 't' va1ur.-, 0'7926 NS j •• • (t! value . 0·4735

NS ; NS, IloIl~lgnifica.nt .

October 1973] CHOLESTEROL CONTE:\'T'OF COW AND BUFFALO GHEE

Table 4. Total cholesterol content of ghee pre:: .. par~d from fore·milk and stripping.'lo of cow

and bulTalo

Total cl1de,terol content (mg%)

Cow' Buffalot

Fore ... Stripo Fore· Strip-milk pjngs milk pings

345 324 311 281 344 326 302 275 343 302 2!H 266 336 297 209 200 33B 315 200 241 337 291 267 243 40& 344 290 271 391 291 343 314 403 2SB 286 249 386 351 296 252 393 344 315 273 385 334 320 290

Average 967·4 324·2 297·4 202·9

• .1 't ' value) 3-9M3·.r; t, <e value, j·94Bl**; significant (PSO· 0 1).

:01; ~

300

280

260

2~0

220

zeo

g \80

~ 160

" ~ 140 .. '" 'Q

"" 120 ()

100

8] 60 50

loT 1- ~ •

2 ~ 4' :; 6 8 9

Days aller parlurilion

fat globule. Baranowski and Rooten berg (1967) also reported a significantly higher cholesterol content of fore-milk than of strippings.

Variations in cholesterol contents of gh .. The free <:holestct'o l content of cow

ghee was 3ignificalltly (P<O' 01) higher than that of buffalo ghee. Variations in free cholesterol content due to season were not stati, tically significant. Average values of free choksterol content in cow and buffalo ghee during winter, autumn, summer ,,,,d ~pring arc given in Table 5.

The esterified choles terol content of cow ghec (average value, 47·6 mg%) was significantly (P<O·OJ) lower than that of bulTalo ghee (average valu!'", 63'5 mg%). This confirms the earlier report (Binda! and Jain, 1972), which was based on a limited number of analyses. Variations (Table 5; Fig. 4) in the esterified choles­terol con tent due to sea~On were highly

~ '" §.

~ J!! .. .!! .2 ()

170

10 L-~-:---!~~-:-~-;:-Q""'" 12345678 9

Days afte( parturi1ion

Fig. 4. Variations in (A) esterified, and (B) free cholesterol content of colostra l fat. I) Cow; 2, buffalo.

923

BINDAL AND JAIN [Vol. ~, No. 10

Tahir:': 5. Total, free and esterified chole.terol content of cow and buffalo ghee in different seasons

Torol cholesterol Free: cholesterol E,terified chol .. tcrol conte" t (mg%) content (mg"lo) content (mg%)

Season Cow Buffalo Cow Buffalo Cow Buflruo

Winter (Dec.-Feb.) 340·8

Spring (March·May) 342·6

Summer (june-August) 315·3

Autumn (Sept.·Nov.) 330·6

------_.

significant (P<O' 01) in both types of ghee, peak value being observed in spring both in cow and buffalo ghee. The esterified cholesterol content of ghec in winter, summer and autumn was significantly lower. Variations in the JeveL~ of esterified cholesterol during winter, summer and autumn wefe not, however, significant.

Variation.' in clwieslerol conltllts of colostral fat

Colostral fat, in the form of DC ghee, was analysed for it~ free and esterified cholesterol contents. There were erratic variations, both in cow and buffalo ghee, during the 9 days following parturition (Fig. 4). However, the estedfied choles­terol content of colostral fat was much higher than that of fat of normal milk.

ACKNOWLEDGEMENTS

The authors thank Dr D. Sundarcsan, Director; and Dr N. C. Ganguli, Dairy Chemist, for their interest in this work. The valua.ble help and guidance towards the statistical planning of the work by S4ri M . Gurnani is very much appreciated.

REFERE!'<CES

AZIM, M. A .• nd MUSTAN'lR, A. J. 1971. Season.l variation!) of cho1esterol in milk of buffa.lo, cow and goat. Dairy Sri. Abstr. 33; 3,10l.

BA.R..ANow-BARANowmI~ S. and ROOTF.NB~RG, S. 1961. Bffbet of pas.ture and stall feeding on

273·3

285·7

773'4

275 · 4

298·B 21S'1 39·0 :H·B

274·0 21l ·3 69·1 77·6

276'0 208'8 40'B 60·8

284·5 210·' 41 ·6 60 ·8 ___ ' __ 0._- _.

cholesterol levels in blood and milk. Do"'" S<i. Ab,/r. 29. 22.

BlNDAL, M. P. and JAm, M. K. 1972. 'Free' and 4estecifil":<P cholesterol in desi ghee prepared from milk of cows and buffaloes. IndilJJt]. Dairy Sci. 25, 236.

BINDAL, M . P. and JAIN, M. K . 1973. Estimation of total cholcsterolin gbee prepared from milk of cows and bu.ffaloe.. ]. Indian cbnn. SIX. SO(I) , 63 .

GULVADY, ·S. L., KANNAN, 1\. and BAlIu, K. P. 1952. Cholesterol content of milk of cOws and buffaloe<. India" J. Da;') Sci. 5, 125.

HOMER, D. R. and VmTANEN, A. 1. 1967. Cow's milk cholesterol. Studie, on the milk of cow. on norma.l and protein~free feew.. MileJrw'· sscruc"'ift 22: I.

r"DlAN STAN(}AJtDS IJ<!TrruTlI. 1966. Me/both of sampling and lut for g"" (lluJ!tf./tJl). 1nd;". ,tandaras. I. S . .' 3508, p. 34. Indian Sta.ndards Institute, New Delhi.

JBN:-l'Es&~ R. and PATrON, S. 1959. Pri/~ci.ples of Dairy Chemistry. p. 268. Wiley Ea.:)tern Pvt., Ltd, New Dc-lhi.

KAYSI!.R", F. and TACHKOV~ D. 1958. The state in which cholesterol exl3 ts in milk. Chern . Abs/r. 52, No. 20716.

LASKOWSKI, K. 1962. Cholesterol conunt of esp, milk a.nd cre'1om in Poland. Dairy Sri . .Abllr. 24: No. 2090.

I ...... x:l41NARAyANA, H. and DA!oTUR, N. N. 1968. Buffaloes.' milk and milk. products. II. Dairy Sci. Ab.,/r. 30, No. 231.

I\oIULDER, H. and ZllrDHO"F, T. A. 1958. The surface layers of milk-fat globJ.les. 4. The cholesterol content (free and tsterified cho­lesterol) of surface layero. N,t/o. Milk Dairy J. 12, 173.

SRJ:.IIVA5A.. .... , rvr. R. and ANANTAlUllSHNANJ C. P. 1964. Milk Produ,/S .f India. p. ' 26. l ndi<\n Coull.cil of l\gdculturat Research. New Delhi.

STEGER, H. 1962. The cholesterol content of the milk of domMtic animal$ during lactation. Dairy Sci. Ab,tr. 2 • • No. 1,451.

924