HPLC-ELSD同时测定瓦布贝母药材中7种异甾体生物碱

Simultaneous Determination of Seven Isosteroidal Alkaloids in Fritillaria wabuensis by HPLC-ELSD Method

目的建立高效液相色谱-蒸发光散射检测法(HPLC-ELSD)同时测定瓦布贝母药材中多种生物碱的含量,为瓦布贝母的质量控制提供方法。方法使用InertSustain C18色谱柱(4.6 mm×250 mm,5μm),以0.1%三氟醋酸(A)-乙腈(B)为流动相进行梯度洗脱。结果本方法专属性好、灵敏度高、简便准确,用于瓦布贝母药材中西贝碱苷、西贝碱、贝母辛、贝母素甲、贝母素乙、异浙贝甲素及川贝酮碱等7种异甾体生物碱的含量测定,线性关系良好(r^2≥0.999 0),加样回收率为90%~105%。结论本方法可检测到几乎所有的贝母类生物碱成分,所得的色谱图可同时作为特征图谱,对不同种类的贝母药材进行鉴别。本条件也适合与质谱检测器联用。异浙贝甲素、川贝酮碱首次通过HPLC在瓦布贝母中被检测到。

OBJECTIVE To establish an effective high performance liquid chromatography coupled with evaporative light scattering detector(HPLC-ELSD)method to simultaneously detect multi alkaloids in Fritillaria wabuensis,which can be used to control the quality of Fritillaria wabuensis.METHODS The method was conducted on an InertSustain C18 column(4.6 mm×250 mm,5μm)with 0.1%TFA(V/V)aqueous solution-acetonitrile as mobile phase eluted in a gradient program.RESULTS The method had good linearity(r^2≥0.999 0).The accuracy was between 90%and 105%.This method was simple,sensitive,accurate and specific,which can be applied to determine imperialine-3-β-D-glucoside,imperialine,peimisine,verticine,verticinone,isoverticine and chuanbeinone in Fritillaria wabuensis.CONCLUSION Nearly all fritillariae alkaloids can be detected under this condition,therefore,the chromatogram can be used as the characteristic chromatogram to testify different kinds of Fritillaria.The method can also be combined with mass spectrometric detector.It′s the first time for isoverticine and chuanbeinone to be detected in Fritillaria wabuensis by HPLC and also the first time for seven isosteroidal alkaloids in Fritillaria wabuensis to be simultaneously determined.