HPLC法测定植物甾醇软胶囊中植物甾醇的含量

QUANTITATIVE ANALYSIS OF PHYTOSTEROLS IN PHYTOSTEROLS SOFTGELS BY AN HPLC METHOD

目的建立测定植物甾醇软胶囊中植物甾醇(以豆甾醇和β-谷甾醇计)含量的HPLC方法,并应用此法测定两种油脂基质功能食品中植物甾醇的含量。方法样品经超声皂化(45℃,45 min)提取后,采用Thermo Accucore C18(4.6 mm×250 mm,5μm)色谱柱以甲醇为流动相进行分离,流速0.8 ml/min,柱温30℃,检测波长205 nm。结果豆甾醇和β-谷甾醇在15min内分离良好,质量浓度分别在5.15~515(r=0.9998)和5.06~506(r=0.9998)μg/ml范围内呈良好线性;分别以两种样品为本底,测得植物甾醇软胶囊A的平均回收率豆甾醇93.6%、β-谷甾醇91.0%,RSD分别为3.2%和2.7%(n=6),3批样植物甾醇含量分别为31.7、31.0、31.0 g/100g;软胶囊B的平均回收率豆甾醇91.7%、β-谷甾醇94.1%,RSD分别为2.2%和2.9%(n=6),3批样植物甾醇含量分别为30.8、31.2、30.2 g/100g。结论该法操作便捷、条件温和、绿色、高效、准确,适用性强,为植物甾醇的分析提供新的途径。

Objective To establish an HPLC method for the determination of phytosterols （stigmasterol and β-sitosterol） in functional foods and quantitatively determine the content of phytosterols in functional foods with two different grease bases. Methods The sample was extracted with saponification by ultrasonic （45℃ , 45 min）. The chromatographic separation was achieved on a Thermo Accucore C18 column （4.6 mm× 250 mm, 5 μm） ,using methanol as the mobile phase at a flow rate of 0.8 ml/min. The column temperature was 30℃, and the detection wavelength was set at 205 nm. Results Stigmasterol and β-sitosterol were well separated within 15 minutes. The calibration curves of stigmasterol and β-sitosterol showed good linearity in the range of 5.15-515 （r = 0.9998） and 5.06-506 （r = 0.9998）μg / mL, respectively. The average recovery in Phytosterols softgel A of stigmasterol and β-sitosterol were 93.6% and 91.0% with RSD of 3.2% and 2.7% （n=6）. Three batches of phytosterols were 31.7, 31.0, 31.0 g/100 g, respectively. The average recovery in softgel B of stigmasterol and β-sitosterol were 91.7% and 94.1% with RSD of 2.2% and 2.9% （n=6）. Three batches of phytosterols were 30.8, 31.2, 30.2 g g/100 g, respectively. Conclusion This method is easy to operate, mild, green, efficient, accurate and adaptable. A new approach for phytosterols analysis by this method is provided.