Economical and Effective Method for Isolation of Cholesterol free Lecithin from Egg Yolk

Amjad Khan

PhD, Scholar, JJT University, Rajasthan

ABSTRACT:

Eggs are the one of the most commonest functional food source of protein, amino acid, lecithin ,oil, vitamins and other vital nutrients. Lecithin forms the major component in the egg yolk, a source of precursors of major Brain Neurotransmitter and other normal body functions. Hence the isolation of cholesterol free lecithin a from an egg yolk by solvent precipitation is essentially important for human use. Cholesterol free lecithin could be the need of the hour with rising cardiac disorders and Brain function disorders. In the present study a simplified, economical and effective method for isolation of cholesterol free lecithin from egg yolk is performed. The yield was found to be significant hence can be used for further studies in field of supplement of foods for brain function.

KEYWORDS: Egg yolk, Functional food, Cholesterol free Lecithin, Solvent precipitation

INTRODUCTION:

Eggs can be classified as a functional food, a hot button for today’s consumer. Definitions of ‘functional foods’ has been put forth by several organizations. According to the Institute of Food Technologists, functional foods provide additional physiological benefit beyond that of meeting basic nutritional needs[1].

They supply all essential amino acids for humans, and provide several vitamins and minerals, including vitamin A, riboflavin (vitamin B2), folic acid (vitamin B9), Vitamin B6,Vitamin B12, Choline, Iron, Calcium, Phosphorus and Potassium. They are also an inexpensive single-food source of protein[2].

Hence due to these facts eggs have always been under study for actives since ages but unfortunately unable to achieve isolation of cholesterol free egg lecithin.

MATERIALS AND METHOD:

There many US patents and research papers related to method for isolation of the said lecithin from the egg yolk these have been established, these are Gustav Klein et al., (1933) US2013804[3], Robert H Sifferd et al., (1945) US2371476 [4], E. Gordon Young et al., (1951)[5], (For Protein fractionation), Yano et al., (1975) US4157404[6], Nath et al., (1976) US3958034[7], Juan f. Santaren et al., (1981)[8] and Hatanaka et al., (1991) US5028449 [9]

Also, for reducing cholesterol in the lecithin, Merchant et al., (1991) US5037661 [10], Merchant et al., (1995) US5378487[11]¬ Merkle et al., (2001) US6217926[12]. Edwin E. Garcia Rojas et al., (2006) [13] , Culy et al., (1994) US5292546 [14] ,Yan Sun et al., (2011) [15].

Other works for reducing cholesterol includes Kijwoski et al., (2000) ,US6093434 [16] and Narabe et al., (1999) US5904945[17]. Modern methods for phospholipids but resisted the cholesterol free lecithin were Emanuele Boselli et al.,(2000) [18] ,Henning Nielsen et al., (2004) [19]

Method development for use of acceptable food additives Akashe et al., (2001) US6235336 [20] , Campbell et al.,(2004) US6773731 [21] , O.J. Catchpole et al., (2008) [22] , J.C. Okonkwo et al., (2009) [23] and Amanda Laca et al., (2010) [24]

In present study Eggs of uniform size were obtained from local market of Hyderguda, Andhra Pradesh supplied by Venkateshwara Hatcheries, Hyderaguda, Andhra Pradesh. The initial weights were noted. All the solvents chemical and reagents used were stand SD Fine Chemicals. The egg’s yolk was separated manually and treated with hydro alcoholic to precipitate the proteins. The hydro alcoholic treated yolk was then treated with acetone and mixed with homogenizer to obtain consistency in the product. and marc (powder)is retained (P1) .This powder obtained was refluxed in reflux condenser using petroleum ether and Choloroform (5:1) as a solvent for 15 minutes. When subjected to filtration. white fine powder was obtained as a residue (P2) .Percentage yield is calculated. A systematic qualitative analysis is thus performed and the lecithin is confirm by fractionation with 6 N HCl followed by partition with petroleum ether. Presence of Cholesterol is detected by Chemical test and TLC for steroids using TLC using Petroleum Ether 60-80 and Methanol (8:2) with Anitmony trichloride spray reagent. Test for presence proteins was also performed using standard.

RESULTS:

The percentage yield of final residue was found to 12.1% w/w of the egg yolk. A systematic qualitative analysis shown in Table No1.On fractionation with 6 N HCl 2 fractions were obtained F1 and F2 results shown in Table No. 2. Thus the obtained residue shows negative test for cholesterol. Absence of Cholesterol is confirmed by results of TLC for steroids. The powder shows the entire negative test for proteins. Hence the residue is indentified as Cholesterol free Lecithin.

Table No. 1

Solvent fraction	Carbohydrate	Glycosides	Flavonoids	Proteins	Steroids
Residue (P2)	-ve	-ve	-ve	-ve	-ve

Table No.2: Phytochemical screening of the residue

Sr. no	Isolation of the egg yolk Lecithin	Average weight(g)	Percentage yield of P2
1.	Residue (P2)	2.61	12.1
2.	Fraction 1 (F1)	1.68	64.36
3.	Fraction 2 (F2)	0.92	35.24
4.	Total F1+ F2	2.6	99.61

F1 and F2 on fractionation using 6 N HCl followed by partition.

DISCUSSION:

The yield obtained from the solvent precipitation of found to maximum of 12.1 %w/w of cholesterol free egg yolk lecithin. The purity of the isolated lecithin (P2) was found to be 99.61%w/w. hence this lecithin can be used for the purpose of various nutraceuticals excipients after performing the basic characterization before accepting and equivalent to standard.

CONCLUSIONS:

Thus the method developed is found to be simple, effective and economical to understand the need of high production for the needs of Indian and Global market.

REFERENCES:

1. Hasler CM, “Functional foods: Their role in disease prevention and health promotion”. Food Technology 52:63–70, (1998).

2. "Food and Agriculture Organization article on eggs". Fao.org.

3. Gustav Klein et al, US Patent 2013804 (1933)

4. Robert H Sifferd et al., US Patent 2371476 (1945)

5. E. Gordon Young et al., “On the Fractionation of the proteins of egg yolk” Journal of Biological Chemistry, June:73-80, (1951)

6. Yano et al., US Patent 4157404 ,(1975)

7. Nath et al., US Patent 3958034 , (1976)

8. Juan f. Santaren M. Rico Ribera “Thermal and MR studies of Chick Embryo Lecithins”Chemistry and Physiics of Lipids, 29:147-155, (1981).

9. Hatanaka et al., US Patent 5028449 , (1991)

10. Merchant et al., US Patent 5037661, (1991)

11. Merchant et al., US Patent 5378487¬ , (1995)

12. Merkle et al., US Patent 6217926,(2001)

13. Edwin E. Garcia Rojas , Jane S. dos Reis Coimbra , Luis A. Minim, Jackson F. Freitas, “Cholesterol removal in liquid egg yolk using high methoxyl pectins”, Carbohydrate Polymers 69 (2007) 72–78

14. Culy et al., US Patent 5292546 , (1994)

15. Yan Sun, Hailing Yang, Xueming Zhong, Ling Zhang, Wu Wang, “Ultrasonic-assisted enzymatic degradation of cholesterol in egg yolk” Innovative Food Science and Emerging Technologies 12 (2011) 505–508

16. Kijwoski et al., US Patent 6093434 , (2000)

17. Narabe et al., US Patent 5904945, (1999)

18. Emanuele Boselli , Maria Fiorenza Caboni, “Supercritical carbon dioxide extraction of phospholipids from dried egg yolk without organic modifier” Journal of Supercritical Fluids; 19 (2000) 45–50

19. Henning Nielsen, Vijai K.S. Shukla, “In situ solid phase extraction of lipids from spray-dried egg yolk by ethanol with subsequent removal of triacylglycerols by cold temperature crystallization” Lebensm.-Wiss. u.-Technology. 37 (2004) 613–618.

20. Akashe et al., US Patent 6235336 ,(2001)

21. Campbell et al., US Patent 6773731,(2004)

22. O.J. Catchpole , S.J. Tallon , J.B. Grey , K. Fletcher , A.J. Fletcher , “Extraction of lipids from a specialist dairy stream” , Journal of Supercritical Fluids; 45 (2008) 314–321

23. J.C. Okonkwo, “Effects of Breed and storage Duration on the Beta-Carotene content of the egg yolk”akistan Journal of Nutrition 8(10):1629-1630, 2009

24. Amanda Laca, Benjamı´n Paredes, Mario Dı´az , “A method of egg yolk fractionation. Characterization of fractions” Food Hydrocolloids 24 (2010) 434–443.

Received on 19.03.2013

Modified on 25.03.2013

Accepted on 05.04.2013

Research Journal of Pharmacognosy and Phytochemistry. 5(2): March-April 2013, 109-110