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

摘要

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 mL/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.