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A new method for simultaneous estimation of unsaponifiable constituents of rice bran oil using HPTLC
Author(s) -
Afinisha Deepam Lawrence S.,
Soban Kumar Dasiah R.,
Sundaresan Andikkannu,
Arumughan Chami
Publication year - 2007
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200700226
Subject(s) - rice bran oil , unsaponifiable , chromatography , squalene , silica gel , wax , chemistry , mathematics , bran , raw material , organic chemistry
Rice bran oil (RBO) is rich in a variety of bioactive phytochemicals otherwise known as unsaponifiable constituents (USC). Oryzanols, phytosterols, tocols, etc . are the major USC in RBO; the methods presently used for their estimation involve different techniques and require pretreatment of the sample. In this paper standardization of a simple method for simultaneous estimation of USC directly from RBO using HPTLC is presented. The method involves a two‐stage separation of USC on a precoated silica gel 60 F 254 TLC plate viz. : TLC‐1 to separate sterols, oryzanols and tocols; TLC‐2 to separate steryl esters, wax, and squalene. Calibration plots using the respective standards were made to determine LOD, LOQ, and linear regression equations. Recovery studies were also conducted and the values ranged from 93.45 to 101.97%. The LOD and LOQ values showed the sensitivity of the method. The instrumental precision was found to be in the range of 0.30 to 1.18 CV%. Quantitative estimation of USC in crude RBO and refined RBO using this method gave a concentration of 52.80 mg/g of USC in the crude and 33.48 mg/g in the refined oil. The present method for estimation of USC using HPTLC is fast, simple, accurate, precise, and sensitive, as demonstrated here.