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目的建立高效液相色谱法(high performance liquid chromatography,HPLC)测定西红花苷Ⅰ和西红花苷Ⅱ含量的分析方法,并对比了不同产地中西红花苷Ⅰ和西红花苷Ⅱ的含量。方法对不同产地西红花样品采用90%乙醇超声提取,提取溶液采用反相色谱柱Ultimate XB-C18(4.6 mm×250 mm,5μm)检测,以乙腈:水=32:68(V:V)(0.1%甲酸)为流动相,流速1.0 m L/min,柱温35℃,检测波长440 nm。结果西红花苷Ⅰ在15~300μg/m L与峰面积积分值成良好的线性关系(r2=0.9996),西红花苷Ⅰ平均加样回收率为96.6%,RSD为1.69%;西红花苷Ⅱ在12~240μg/m L范围内与峰面积积分值成良好的线性关系(r2=0.9998),西红花苷Ⅱ平均加样回收率为96.1%,RSD为1.58%。不同西红花由于产地和种植不同,西红花苷Ⅰ和西红花苷Ⅱ含量差异较为明显。结论该方法简单、灵敏、稳定、可靠,可用于不同西红花中西红花苷Ⅰ和西红花苷Ⅱ的含量测定。
OBJECTIVE To establish a method for the determination of crocin Ⅰ and crocin Ⅱ by high performance liquid chromatography (HPLC), and to compare the contents of crocin Ⅰ and crocin Ⅱ in different areas . Methods The saffron samples from different areas were extracted with 90% ethanol by ultrasonic. The extraction solution was detected on a reversed-phase column Ultimate XB-C18 (4.6 mm × 250 mm, 5 μm) using acetonitrile: water = 32:68 (0.1% formic acid) as mobile phase at a flow rate of 1.0 mL / min and a column temperature of 35 ℃ with a detection wavelength of 440 nm. Results The cercariain Ⅰ had a good linear relationship with the peak area integral value (r2 = 0.9996) at 15-300μg / mL and the average recovery of crocin Ⅰ was 96.6% and the RSD was 1.69% The linear relationship between peak area and concentration of glycoside Ⅱ (r2 = 0.9998) was 12 ~ 240μg / mL. The average recoveries of crocin Ⅱ was 96.1% and the RSD was 1.58%. Different crocus due to different producing areas and planting, crocin Ⅰ and crocin Ⅱ content more obvious differences. Conclusion The method is simple, sensitive, stable and reliable and can be used for the determination of crocin Ⅰ and crocin Ⅱ in different crocus flowers.