四种常见保健药材中16种真菌毒素的测定和污染状况研究

Determination and contamination status of 16 mycotoxins in four common health medicinal materials

目的建立同位素内标-QuEChERS-超高压液相色谱-串联质谱测定常见保健药材三七、西洋参、天麻和丹参中16种真菌毒素的检测方法,研究药材本身及日常保存中真菌毒素的存在状况。方法样品中添加内标,用乙腈-水-甲酸(70∶29∶1,体积比)溶液提取,离心,上清液经QuEChERS材料净化,多反应监测模式测定,基质匹配内标法定量。对色谱与质谱条件、提取溶剂和净化材料进行优化,并对方法学进行考察。利用该方法对18份实际样品初采及储藏1年后分别进行测定,并对测定结果进行分析。结果16种真菌毒素在各自的线性范围内具有良好线性(r^(2)=0.994~0.999),检出限和定量限分别为(LODs=0.01~15μg/kg,LOQs=0.03~40μg/kg),高低两浓度加标水平下所得的平均加样回收率为83.2%~103.1%,精密度在1.8%~8.1%之间。在原药材和保存1年后药材检出不同含量的多种真菌毒素,主要存在曲霉的污染。结论该方法简便、安全、灵敏和准确率高,适用于同类药材的高通量检测。三七、西洋参、天麻和丹参四种药材在居家环境下易受到霉菌污染,不适于长期储存。

Objective To establish a method for the determination of 16 mycotoxins in four common health medicinal materials:Panax notoginseng,American ginseng,Gastrodia elata and Salvia miltiorrhiza by isotope internal standard-QuEChERS-ultra-high pressure liquid chromatography-tandem mass spectrometry was established and to investigate the existence of mycotoxins in the medicinal materials and daily preservation.Methods The samples were added with internal standard,extracted with acetonitrile-water-formic acid(70∶29∶1,v/v)solution,centrifugated,purified with QuEChERS material,determined by multiple reaction monitoring mode,and quantified with matrix-matched internal standard method.The treated sample solution was determined by multi reaction monitoring mode and quantified by matrix matching internal standard method.The conditions of chromatography and mass spectrometry,extraction solvent and purification materials were optimized.The method was validated.Then the method was used to determine 18 real samples after initial collection and after one-year storage respectively,and the determination results were analyzed.Results The linearity of 16 mycotoxins was good in their respective linear range(r^(2)=0.994~0.999).The detection limits(LODs)and quantitative limits(LOQs)were 0.01~15μg/kg and 0.03~40μg/kg,respectively.Under the two spiked sample level of high and low concentrations,the average spiked recoveries were 83.2%~103.1%and the precisions were 1.8%~8.1%.A variety of mycotoxins with different contents were detected in the original medicinal materials and one-year stored materials,mainly contaminated by Aspergillus.Conclusion The established method is simple,safe,sensitive,and accurate.It is suitable for the high-throughput detection of similar medicinal materials.Panax notoginseng,American ginseng,Gastrodia elata and Salvia miltiorrhiza are vulnerable to mould pollution in the home environment and were not suitable for long-term storage.