New Researches in Biotechnology - Facultatea de Biotehnologii ...
New Researches in Biotechnology - Facultatea de Biotehnologii ... New Researches in Biotechnology - Facultatea de Biotehnologii ...
184Proceeding of the 4 rd International Symposium“NEW RESEARCH IN BIOTECHNOLOGY” USAMV Bucharest, Romania, 2011CAROTENE CONTENT FROM MILK SAMPLES OBTAINED INDIFFERENT FEEDING CONDITIONSFULVIA A. MANOLACHE 1 , DENISA E. DUŢĂ 1 , VALENTIN IONESCU 1 , LUMINITACATANA 1 , VASILE A. BLAJ 2 , AMALIA MITELUŢ 31 National Institute of Research&Development for Food Bioresources – IBA Bucharest, hfulvia@yahoo.com,denisa_duta@yahoo.com, lumi_catana@yahoo.co.uk, valentin.ionescu@bioresurse.ro2Grassland Research and Development Institute, Braşov, blajadi@yahoo.com3 University of Agronomic Sciences and Veterinary Medicine Bucharest -Faculty of Biotechnology,amalia@agral.usamv.roAbstract. Carotene is important in human health and nutrition, because is a precursor to vitamin A.In spite of lower percentage in milk, carotene is involved in the sensorial properties of dairy productsthrough their colorant and antioxidant properties. The aim of this research was to study thevariability of carotene content according to farm feeding practices. Carotene content was determinedusing UV-VIS 550 spectrophotometer. Milk samples were analyzed from physico-chemical point ofview. The metal content (Cu, Fe and Zn) of each sample was determined by Atomic AbsortionSpectrophotometer (Analyst 400). The obtained results showed a variation of beta carotene contentaccording to the feeding mode of cows.Keywords: milk, carotene, spectrophotometer, Atomic Absorbtion1. INTRODUCTIONCarotenoids are notable for their wide distribution, structural diversity, and variousfunctions. Animals are incapable of carotenoid biosynthesis, and hence depend on dietarycarotenoids, which are selectively or unselectively absorbed, converted to vitamin A.[4]Even it is obviously in several countries that carotene contribution in human diet iscoming in large percentage from carrots, as well as from other fruits and vegetables, asmaller concentration is available from diary products, giving a better position to milk inconsumer acknowledge. More, the carotenes may play a rol in stabilising the oxidablecompounds of milk. Despite their benefic roles, a special attention should be paid tocarotene transfer from feeding to cow milk.[3]Data from literature shows about 10 identified carotenes in feed (ex. Carotene,xantofile), their concentration depending on development status and their conservationperiod. The sensitivity of β-carotene during rumination is depending a lot of the feedingsource. From the class of ruminants, only cattle accumulates high concentration ofcarotenes, especially β-carotene, most probably due to the low efficiency of vitamin Asynthesis in enterocytes. Carotenes from cow milk are mostly β-carotenes trans and, inlower concentration, lutein. In milk, the concentration varies more in the case of β-carotenethan for retinoic those plasmatic concentration is well fixed. The β-carotene concentrationin milk is depending on ditetic suplements used. The animal and the factors related to himis affecting the milk production (i.e race).[2]In this article the presented research results are concerning the level of β-carotenefrom milk of the cows grew in plain and mountain areas, differentiated fed due to season.
Proceeding of the 4 rd International Symposium“NEW RESEARCH IN BIOTECHNOLOGY” USAMV Bucharest, Romania, 20112. MATERIALS AND METHODS2.1 SAMPLES OF MILKFor completing the experiments, milk samples from two areas have been analyzed: fromplain area and from mountain area from cows fed with different types of feed. 11 milksamples of cows fed both summer and winter periods from plain area of IBNA Balotestihave been analyzed. From the mountain area of Blana Bucegi milk samples of cows grazingon different pasture have been analyzed.Table 1. Milk samples used in experimentsSample cod Feeding mode OriginP1,P2, P3 combined fodder, hay - alfalfa, corn silage IBNA BalotestiA1,A2,A3C1,C2,C3D1,D2,D3E1,E2,E3natural pasture fertilizednatural pastures, limed, chemical fertilised,padockedsown pasture, limed, chemical fertilised,padockednatural pastures near the experimental fieldexperimentalfieldsBlana Bucegi(1800 m)TI,T2,T3natural pastures from the experimental fieldP4,P5,P6,P7,P8,P9,P10,P11 combined fodder, green fodder IBNA BalotestiMilk samples A1..3; C1..3 ; D1..3; E1..3; T1..3 have been taken in august from 15cows fed on 5 different pastures each of 0,75 ha. On each pastures 3 cows have been fedabout 85 days on summer time. The pastures differ on type, level and fertilization level aswell as the exploiting way. Before analysis, the samples have been frozen at -18 0 C.2.2 METHODSUV-VISThe concentration of β-carotene from the considered milk samples has been determinedwith UV-VIS 550 spectrophotometer. The milk samples have been thawed, heated at roomtemperature, homogenized and then analyzed. Saponification of fat from milk has beendone at cold with hydro-alcoholic solution of potassium hydroxide and carotene extractionwas done with n-hexane. The extract purification has been done on chromatographiccolumn (activated aluminum oxyde and anhydrous sodium sulphate). The β-caroteneextract colouring intensity in hexane has been measured with the spectrophotometer (λ=450nm, the reference sample was n-hexane).The content of β-carotene, in µg at 100 ml of product, was calculated with the followingformula:C × VVβ-carotene = 1× 100[µg/100ml]of which:C – the concentration of β-carotene, red on calibration curve in µg/mlV 1 – the extract volume of β-carotene in hexane, in ml185
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Proceed<strong>in</strong>g of the 4 rd International Symposium“NEW RESEARCH IN BIOTECHNOLOGY” USAMV Bucharest, Romania, 20112. MATERIALS AND METHODS2.1 SAMPLES OF MILKFor complet<strong>in</strong>g the experiments, milk samples from two areas have been analyzed: frompla<strong>in</strong> area and from mounta<strong>in</strong> area from cows fed with different types of feed. 11 milksamples of cows fed both summer and w<strong>in</strong>ter periods from pla<strong>in</strong> area of IBNA Balotestihave been analyzed. From the mounta<strong>in</strong> area of Blana Bucegi milk samples of cows graz<strong>in</strong>gon different pasture have been analyzed.Table 1. Milk samples used <strong>in</strong> experimentsSample cod Feed<strong>in</strong>g mo<strong>de</strong> Orig<strong>in</strong>P1,P2, P3 comb<strong>in</strong>ed fod<strong>de</strong>r, hay - alfalfa, corn silage IBNA BalotestiA1,A2,A3C1,C2,C3D1,D2,D3E1,E2,E3natural pasture fertilizednatural pastures, limed, chemical fertilised,padockedsown pasture, limed, chemical fertilised,padockednatural pastures near the experimental fiel<strong>de</strong>xperimentalfieldsBlana Bucegi(1800 m)TI,T2,T3natural pastures from the experimental fieldP4,P5,P6,P7,P8,P9,P10,P11 comb<strong>in</strong>ed fod<strong>de</strong>r, green fod<strong>de</strong>r IBNA BalotestiMilk samples A1..3; C1..3 ; D1..3; E1..3; T1..3 have been taken <strong>in</strong> august from 15cows fed on 5 different pastures each of 0,75 ha. On each pastures 3 cows have been fedabout 85 days on summer time. The pastures differ on type, level and fertilization level aswell as the exploit<strong>in</strong>g way. Before analysis, the samples have been frozen at -18 0 C.2.2 METHODSUV-VISThe concentration of β-carotene from the consi<strong>de</strong>red milk samples has been <strong>de</strong>term<strong>in</strong>edwith UV-VIS 550 spectrophotometer. The milk samples have been thawed, heated at roomtemperature, homogenized and then analyzed. Saponification of fat from milk has beendone at cold with hydro-alcoholic solution of potassium hydroxi<strong>de</strong> and carotene extractionwas done with n-hexane. The extract purification has been done on chromatographiccolumn (activated alum<strong>in</strong>um oxy<strong>de</strong> and anhydrous sodium sulphate). The β-caroteneextract colour<strong>in</strong>g <strong>in</strong>tensity <strong>in</strong> hexane has been measured with the spectrophotometer (λ=450nm, the reference sample was n-hexane).The content of β-carotene, <strong>in</strong> µg at 100 ml of product, was calculated with the follow<strong>in</strong>gformula:C × VVβ-carotene = 1× 100[µg/100ml]of which:C – the concentration of β-carotene, red on calibration curve <strong>in</strong> µg/mlV 1 – the extract volume of β-carotene <strong>in</strong> hexane, <strong>in</strong> ml185
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