手性高效液相色谱法测定板蓝根中表告依春和告依春含量

Determination of epigoitrin and goitrin in Isatidis Radix by chiral high performance liquid chromatography

采用Chiralpak IC手性柱(250mm×4.6mm,5μm),建立了正相高效液相色谱测定板蓝根中表告依春(R-告依春)和告依春(S-告依春)含量的方法。考察了流动相组成、流速和柱温对分离度的影响。经优化后的实验条件:以正己烷-异丙醇(体积比为90∶10)为流动相,流速为0.8mL/min;检测波长为245nm,柱温为20℃。在此优化条件下,表告依春和告依春分离度为3.4,检出限为2.0mg/L,在0.02～2.0g/L范围内有良好的线性关系,平均回收率为101%,相对标准偏差(RSD)小于3.0%(n=6)。本方法可将具有抗病毒活性的表告依春与其对映体告依春基线分离并测定,专属性强,能有效控制板蓝根的质量。

A Chiralpak IC column （250 mm×4.6 mm,5μm） was used to establish normal phase high performance liquid chromatographic （NP-HPLC） method for the chiral separation of epigoitrin （R-goitrin） and goitrin （S-goitrin） in Isatidis Radix. The composition of mobile phase,flow rate and column temperature were optimized to get better resolution. The final chromatographic conditions were established as follows：n-hexane-isopropanol （90：10,v/v） as the mobile phase with a flow rate of 0.8 mL/min,the detection wavelength at 245 nm and the column temperature at 20 ℃. Under the above optimized chromatographic conditions,the resolution of 3.4 was achieved for epigoitrin and goitrin. The limit of detection （LOD） was 2.0 mg/L. The linear range was 0.02-2.0 g/L. The average recovery was 101% with the relative standard deviation （RSD） lower than 3.0% （n=6）. The proposed method was exclusive by determining the antivirus epigoitrin and its enantiomer separately,which provided the effective quality control of Isatidis Radix.