目的 建立免疫親和柱前處理、超高效液相色譜串聯(lián)熒光檢測器（UPLC-FLD）分析，同時用于檢測逍遙片原料藥材柴胡、當(dāng)歸、炙甘草、炒白術(shù)、白芍及中間提取物1和2中赭曲霉毒素A （ochratoxin A，OTA）的方法。方法 樣品用80%甲醇-水溶液（體積比）超聲提取，離心，免疫親和柱純化，ACQUITY UPLC BEH C₁₈色譜柱分離，流動相A為0.5%冰醋酸溶液（pH值為2.97），流動相B為乙腈，梯度洗脫。激發(fā)波長310 nm，發(fā)射波長465 nm，柱溫30℃，體積流量0.3 mL/min，進(jìn)樣體積2 μL，熒光檢測器檢測。結(jié)果 OTA在1.5~37.5 ng/mL顯示良好的線性關(guān)系，R² ≥ 0.999 1。OTA加標(biāo)回收率為92.36%~119.89%，RSD為2.50%~9.12%。樣品在48 h內(nèi)穩(wěn)定。共檢測42批次樣品，33批呈陽性，柴胡和炙甘草被污染。結(jié)論 建立的免疫親和柱前處理、UPLC檢測的方法可用于中藥和中間提取物中OTA的檢測，排除假陽性結(jié)果，適合痕量分析，結(jié)果準(zhǔn)確可靠。

Objective To establish an immunoaffinity pre-column processing, high performance liquid chromatography tandem fluorescence detector analysis, and to detect the raw materials of Xiaoyao Tablets Bupleuri Radix, Glycyrrhizae Radix Et Rhizoma, Radix Paeoniae Alba, Rhizoma Atractylodis Macrocephalae, Radix Angelicae Sinensis and Intermediate Extract 1, Extract 2 Method of Ochratoxin A. Method The sample was extracted with 80% methanol-water solution (volume ratio) ultrasonically, centrifuged, and purified by immunoaffinity column, separated by ACQUITY UPLC BEH C₁₈ column, mobile phase A:0.5% glacial acetic acidwater solution (pH=2.97), mobile phase B:Acetonitrile elution, excitation wavelength 310 nm, emission wavelength 465nm, column temperature 30℃, flow rate 0.3 mL/min, injection volume 2 μL, fluorescence detector detection. Results Ochratoxin A showed a good linear relationship in the mass concentration range of 1.5 to 37.5 μg/mL, and R² was greater than 0.999 1. The recovery rate of ochratoxin A was 92.36%-119.89%, and the relative standard deviation was 2.50%-9.12%. The sample is stable within 48 hours. A total of 42 batches of samples were detected and 33 batches were positive. Bupleuri Radix and Glycyrrhizae Radix Et Rhizoma were contaminated. Conclusion The established immunoaffinity column pretreatment and ultra-high performance liquid chromatography detection method is suitable for the detection method of OTA in traditional Chinese medicine and intermediate extracts, eliminating false positive results,suitable for trace analysis,conforming to international reporting standards,and the results are accurate and reliable.

R917

國家科技部“重大新藥創(chuàng)制”科技重大專項（2018ZX09303024）