西红花中西红花苷Ⅰ和苷Ⅱ的含量测定

Determination of crocusⅠand crocusⅡin saffron

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 develop a method for determination of crocusⅠ and crocusⅡ in saffron by high performance liquid chromatography (HPLC), and compare the content of crocusⅠand crocusⅡ in different saffron.MethodsThe saffron was extracted by ultrasonic in 90% ethanol solution. Separations of extraction solution were carried out on an Ultimate XB-C18(4.6 mm×250 mm, 5μm). The mobile phase(volume rate)was ACN:H2O = 32:68 (V:V) (0.1% methanoic acid), the flow was 1.0 mL/min, column temperature was 35℃, and the detection wavelength was 440 nm.ResultsCrocusⅠ had a good linearity with the peak areas in 15~300μg/mL (r2=0.9996). The average recovery was 96.6% and the RSD was 1.69%. CrocusⅡ had a good linearity with the peak areas in 12~240μg/mL(r2=0.9998). The average recovery was 96.1% and the RSD was 1.58%. The difference of content of crocusⅠ and crocusⅡ was obvious in different cultivated fields.ConclusionThis method is simple, sensitive, specific, and reproducible, which can be used as the method for measuring the content of crocusⅠ and crocusⅡ in saffron.