25种常用中药饮片常见真菌毒素污染分布比较研究

Comparative Study on the Distribution of Mycotoxins Contamination in 25 Kinds of Common Herbal Decoction Pieces

关键词:  中药饮片  真菌毒素  超高效液相色谱三重四极杆质谱法

Key words:  herbal decoction pieces  mycotoxins  UPLC-Q-TOF-MS/MS

摘要:

目的 建立中药饮片10种真菌毒素的超高效液相色谱三重四极杆质谱联用方法,并对市售25批中药饮片的真菌毒素污染分布比较研究。方法 样品经70%甲醇超声处理,HLB固相萃取柱净化,Waters ACQUITY UPLC^®BEH C₁₈(2.1 mm×100 mm,1.7 μm)分离,流动相为乙腈-甲醇(1:1)和0.01%甲酸水溶液梯度洗脱,ESI离子源正离子扫描,通过多反应检测模式测定,外标法定量。结果 10种真菌毒素在0.31~250 ng·mL^-1内线性关系良好,相关系数均>0.991,平均加样回收率为83.60%~131.80%,RSD为2.35%~7.85%,10种真菌毒素的检出限为0.14~16.30 ng·kg^-1。25批中药材中,伏马毒素B2检出率最高(28%),其次为呕吐毒素(20%);黄曲霉素B2等4种真菌毒素未检出;果实种子类真菌毒素污染最严重(66.7%),动物类药材真菌毒素污染率最低(20%)。结论 本方法可用于中药饮片中10种真菌毒素的检测;除了黄曲霉毒素外,对于中药饮片中其他真菌毒素应加强监测,其中尤其应注意果实种子类中药的真菌毒素监测。

Abstract:

OBJECTIVE To establish a method for the simultaneous determination of 10 kinds of mycotoxins in herbal decoction pieces by ultra-performance liquid chromatography-tandem mass spectrometry, and to compare the distribution of mycotoxins contamination in 25 batches of herbal decoction piece. METHODS The samples were treated by ultrasonic treatment with 70% methanol, purified by HLB column, and separated by Waters ACQUITY UPLC^® BEH C₁₈(2.1 mm×100 mm, 1.7 μm). The mobile phase consisted of acetonitrile-methanol(1:1) and 0.01% formic acid solution gradient elution. Multiple response monitoring in ESI ion source positive ion mode and quantitative by external standard method. RESULTS The linear ranges of the 10 mycotoxins were 0.31-250 ng·mL^-1 with correlation coefficients greater than 0.991. The average recoveries were 83.60%-131.80%, the RSD were 2.35%-7.85%, and the detection limits of the 10 mycotoxins were 0.14-16.30 ng·kg^-1. Among 25 batches of herbal decoction pieces, fumonisin B2 had the highest detection rate(28%), followed by deoxynivalenol(20%), and four mycotoxins such as aflatoxin B2 and so on were not detected. Mycotoxin contamination rate of fruit and seed was the most serious(66.7%), and that of animal medicinal materials was the lowest(20%). CONCLUSION This method can be used for the detection of 10 mycotoxins in herbal decoction pieces. In addition to aflatoxin, the monitoring of other mycotoxins in herbal decoction pieces should be strengthened, especially the monitoring of mycotoxins in fruits and seeds.