Abstract
An efficient, fast, and reliable reversed phase high-performance liquid chromatography based method was developed to detect specific adulterant in premium and pure ghee (clarified butterfat) samples. The method is based on the detection of cholesterol and β-sitosterol as markers in the unsaponifiable matter of pure ghee and adulterated ghee samples, respectively. The reversed phase high-performance liquid chromatography method resolved cholesterol, stigmasterol, campasterol, and β-sitosterol at Rt values of 17.681 ± 0.122, 19.7 ± 0.2, 20.2 ± 0.2, and 22.7 ± 0.2 min., respectively. β-sitosterol served as an indicator for the adulteration in ghee by plant and specific adulterant oil at dose dependent manner (1–5 g/100 g of ghee). Validation of the method revealed that it was fast, economical, highly reliable, and comparable with the reversed-phase thin layer chromatography method with no false negatives.
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