免疫亲和柱层析净化-高效液相色谱法检测中药材中赭曲霉毒素A

Determination of Ochratoxin A Content in Chinese Medicinal Materials by Immunoaffinity Column Chromatography Purification-HPLC

目的建立中药材赭曲霉毒素A（OTA）污染检测的高效液相色谱（HPLC）法。方法经打粉的药材用甲醇-水（80∶20）提取,经OTA免疫亲和柱层析净化,测定。色谱柱为Agilent Zorbax SB-C18柱（250 mm×4.60 mm,5μm）,流动相为乙腈-水-乙酸（49.5∶49.5∶1.0）,流速为1 m L/min,荧光检测器检测,激发波长为333 nm,发射波长为460 nm,柱温为35℃,样品池温度为10℃,进样量为20μL。结果色谱峰面积与质量浓度之间有良好的线性关系,方法的检出限为0.2 ng/g,加样回收率为64.4%~106%,RSD为0~28.5%（n=3）。结论该方法操作简便、准确,重复性好,可用于中药材及中成药等药材中赭曲霉毒素A的测定。

Objective To establish a HPLC method for the determination of ochratoxin A（OTA）contamination in the Chinese medicinal ma-terials. Methods The powdered medicinal materials were extracted by methanol-water（80 ：20）,purified by the OTA immunoaffinity column and determined. The chromatographic column was with the Agilent Zorbax SB-C18 column（250 mm × 4. 60 mm,5 μm）,the mobile phase was acetonitrile-water-acetic acid（49. 5 ：49. 5 ：1. 0）,the flow rate was 1 mL/min,detected by the fluorescence detector,the excitation wave-length was 333 nm and the emission wavelength was 460 nm,the column temperature was 35 ℃,the sample temperature was 10 ℃ and the sample volume was 20 μL. Results The area of chromatographic peak and the concentration demonstrated a good linear relationship,its de-tection limit was 0. 2 ng/g,the recovery rate of samples was 64. 4% -106%,RSD was 0-28. 5%（ n=3）. Conclusion This method is sim-ple,accurate,better reproducible,which can be used for the determination of OTA in the Chinese medicinal materials.