DEVELOPMENT AND VALIDATION OF A REVERSED PHASE-PLANAR CHROMATOGRAPHY (HPTLC) METHOD FOR DETERMINATION OF BHT AND ITS APPLICATION IN FOOD PRODUCTS
Objective: Development and validation of a RP-HPTLC method coupled with densitometry analysis for the determination of BHT as such and in commercial food sample (Butter). Methods: Chromatographic separation of BHT was achieved by employing a mobile phase consisting of Methanol â€“ Water (9.5:0.5) on precoated aluminium TLC plates, silica gel RP-18 F254sand UV densitometric quantitative evaluation was performed by measuring the absorbance reflectance of the analyte spots at 254Â nm. Results: The BHT was satisfactorily resolved with Rf value 0.48 Â± 0.02. The study was carried out in the range of 500 ng to 5000 ng. The LOQ and LOD for BHT were 0.524 and 0.157 ng respectively. The method was validated with respect to linearity, repeatability, specificity, accuracy (recovery) and precision. Conclusion: The proposed method has high degree of repeatability and will provide fast and cost-effective quantitative control for routine analysis of BHT as single and in food sample.
Keywords: BHT, RP-HPTLC, Densitometry, Planar chromatography, Food product, Method Development and Validation
2. Nishaa S, Vishnupriya M, Sasikumar JM, Hephzibah PC, Gopalakrishnan VK. Antioxidant activity of ethanolic extract of Maranta arundinacea L. tuberous rhizomes. Asian J Pharm Clin Res 2012; 5(4): 85-8.
3. Sindhu S, Manorama S. Screening of Polycarpaea corymbosa lam. (Caryophylaceae) for its in vitro antioxidant activity. Asian J Pharm Clin Res 2012; 5(4): 175-8.
4. Nobuyuki I, Shoji F, Hiroyuki T. Carcinogenicity and modification of the carcinogenic response by BHA, BHT and other antioxidants. Critical Rev in Toxicology 1985; 15(2):109-50.
5. Olsen P, Meyer O, Bille N, WÃ¼rtzen G. Carcinogenicity study on butylated hydroxytoluene (BHT) in Wistar rats exposed in utero. Food and Chemical Toxicology 1986; 24(1):1â€“12.
6. Witschi H. Enhanced tumour development by butylated hydroxytoluene (BHT) in the liver, lung and gastro-intestinal tract. Food and Chemical Toxicology 1986; 24(10-11):1127 -30.
7. http://en.wikipedia.org/wiki/Butylated_hydroxytoluene: visited on 20.06.2012.
8. Williams G, Iatropoulos M, Whysner J. Safety assessment of butylated hydroxyanisole and butylated hydroxytoluene as antioxidant food additives. Food and Chemical Toxicology 1999; 37(9-10); 1027â€“38.
9. http://apps.kemi.se/flodessok/floden/kemamne_Eng/BHT_eng.htm: Visited on 18.06.2013.
10. Mohammed A, ZainiBin A, Fasihuddin B. Optimization and validation of RP-HPLC-UV/Vis method for determination phenolic compounds in several personal care products. International Journal of Analytical Chemistry, vol. 2011, Article ID 858153, 9 pages, 2011. Doi: 10.1155/2011/858153.
11. Miao C, Qinghai X, Mousheng L, Yaling Y. Cloud-point extraction and reversed-phase high-performance liquid chromatography for the determination of synthetic phenolic antioxidants in edible oils. J Food Sci 2011; 76(1):98-103.
12. Lee M, Lin C, Li Z, Tsai T. Simultaneous analysis of antioxidants and preservatives in cosmetics by supercritical fluid extraction combined with liquid chromatography-mass spectrometry. J Chromatogr A 2006; 1120(1-2):244-51.
13. Sumiko T, Maki N, Hisaya T, Yukio T, Yasuhide T. Determination and confirmation of five phenolic antioxidants in foods by LC/MS and GC/MS. Shokuhin Eiseigaku Zasshi 2005 ; 46(3): 63-71.
14. Youk-meng C, Hsiu-jung L. Gas Chromatographic determination of synthetic antioxidants in edible fats and oils â€“ a simple methylation method. J of Food and Drug Anal 2001; 9(1):20-6.
15. Hsiu-jung L, Mei-lin W, Youk-meng C, Chung-wen C, Bao S, Shyr-lin T, Lung-chi C, Min-hua Y. Effects of extraction solvent on gas chromatographic quantitation of BHT and BHA in chewing gum. J of Food and Drug Anal 2003; 11(2):141-7.
16. Zhang W, Wu C, Wang C, Yang Z, Liu L. Determination of antioxidants butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) in cosmetics by gas chromatography-mass spectrometry selected ion method. Se Pu 2002; 20(2):178-81.
17. Min-Hua Y, Hsiu-Jung L, Youk-Meng C. A rapid gas chromatographic method for direct determination of BHA, BHT and TBHQ in edible oils and fats. Food Research Int 2002; 35(7):627-33.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.