基于《中华人民共和国药典》优化丹参中33种禁用农药残留物检测的前处理方法

Optimization of the Pretreatment Methods for Detecting 33 Forbidden Pesticide Residues in Salvia miltiorrhiza Based on Pharmacopoeia of The People's Republic of China

目的以丹参为研究对象,比较2020年版《中华人民共和国药典》中3类前处理方法对33种禁用农药残留物回收率的影响,进一步优化快速样品处理法(quick,easy,cheap,effective,rugged and safe,QuEChERS)。方法分别从基质效应和回收率方面比较《中华人民共和国药典》3类前处理方法,并对QuEChERS法进行研究与优化,样品加水后由1%醋酸-乙腈提取,依次加入QuEChERS盐包与QuEChERS净化管盐析净化,提取液经浓缩后分别加入水和内标磷酸三苯酯,结合液相色谱-串联质谱法和气相色谱-串联质谱法以添加水平为定量限浓度水平测定样品的回收率和RSD。结果采用《中华人民共和国药典》前处理方法时,丹参中33种禁用农药(54个单体)在添加水平为定量限浓度水平时,部分农药回收率较低;优化QuEChERS法后,丹参中33种禁用农药(54个单体)在一定浓度下,线性关系良好,r均大于0.990,其回收率为64.43%~127.35%时,RSD均<15%;199批饮片和60批药材中共检出2种农药,甲拌磷检出率为3.86%,克百威检出率为1.93%,但残留量均未超过2020年版《中华人民共和国药典》规定限度。结论优化后的QuEChERS法快速简便,有针对性地对丹参中33种禁用农药残留进行准确检测,对于丹参的生产种植与日常监控具有一定的参考意义。

Objective To investigate the effect of the three pretreatment methods in Pharmacopoeia of The People's Republic of China(2020 edition)on the recovery rate of 33 forbidden pesticide residues with Salvia miltiorrhiza as the research object,and to further optimize the quick,easy,cheap,effective,rugged,and safe(QuEChERS)method for sample treatment.Methods The three pretreatment methods in Pharmacopoeia of The People's Republic of China were compared in terms of matrix effect and recovery rate,and the QuEChERS method was studied and optimized.The sample was extracted using 1%acetic acid-acetonitrile after water was added,and then QuEChERS salt packet and QuEChERS purification tube were used for salting-out and purification.After concentration,the extracting solution was added with water and the internal standard triphenyl phosphate,respectively,and liquid chromatography-tandem mass spectrometry and gas chromatography-tandem mass spectrometry were used to measure the recovery rate and relative standard deviation(RSD)of the sample with the addition level as the limit of quantification(LOQ).Results When the pretreatment methods in Pharmacopoeia of The People's Republic of China were used,some of the 33 forbidden pesticides(54 monomers)in Salvia miltiorrhiza had a relatively low recovery rate at the addition level of LOQ.After the QuEChERS method was optimized,the 33 forbidden pesticides(54 monomers)in Salvia miltiorrhiza had a good linear relationship at a certain concentration(r>0.990),with a recovery rate of 64.43%-127.35%(RSD<15%).Two pesticides were detected in 199 batches of decoction pieces and 60 batches of medicinal material,and the detection rates of phorate and carbofuran were 3.86%and 1.93%,respectively,but the residues did not exceed the limits specified in Pharmacopoeia of The People's Republic of China(2020 edition).Conclusion The optimized QuEChERS method is rapid and simple and can be used for accurate detection of 33 forbidden pesticide residues in Salvia miltiorrhiza,which provides a certain reference for the production,planting,and daily monitoring of Salvia miltiorrhiza.