Phytosterol Determination and Method Validation for Selected Nuts and Seeds
2017
Islam, Md Atiqual | Jeong, Beom-Gyun | Jung, Jiyoung | Sin, Ŭi-chʻŏl | Choi, Sung-Gil | Chun, Jiyeon
A method involving alkali and/or acid hydrolysis of phytosterols followed by trimethylsilyl ether derivatization coupled with GC-FID analysis was validated and applied in the analysis of major phytosterols (campesterol, stigmasterol, β-sitosterol, and Δ⁵-avenasterol) in nuts (n = 7), seeds (n = 9), legumes (n = 2), and grain (n = 1). The acid-labile Δ⁵-avenasterol was extracted with alkaline hydrolysis only before derivatization. Quantification of all phytosterols was done using the computed relative response factor of 5α-cholestane (internal standard). Analyses of internal and external phytosterol standards showed good linearity for all phytosterols (R ² of 0.999); LOD and LOQ of phytosterols were determined to be 0.01–0.12 and 0.04–0.40 mg/100 g, respectively. Repeatability and reproducibility precision analyses showed acceptable coefficient of variation of less than 3 and 4%, respectively, and satisfactory Horwitz ratio values of <1.0. Excellent accuracy was proved by the high recovery values of 91.4–106.0% for campesterol, β-sitosterol, and stigmasterol. Δ⁵-Avenasterol, the most oxidation-susceptible sterol, showed a recovery of about 60%. The total phytosterol (sum of major phytosterols quantified) contents in the 19 samples varied from 38.8 mg/100 g (white quinoa seed) to 246.2 mg/100 g (sunflower seed). β-Sitosterol was the predominant phytosterol (54–86.0% of total) among all samples except fennel seed in which stigmasterol was predominant. Analytical quality control chart maintained during the study period showed that assays were performed under control. Method validation indicated that the analytical method can be applied for accurate determination of campesterol, β-sitosterol, and stigmasterol in selected food samples.
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