超高效液相色谱-串联质谱法同时测定抗(抑)菌类消毒产品中苯海拉明和利多卡因的含量

Simultaneous Determination of Diphenhydramine and Lidocaine in Antifungal Disinfection Products by Ultra-High Performance Liquid Chromatography Tandem Mass Spectrometry

取抗(抑)菌类消毒产品样品0.200 g,加入1.00 mg·L^(-1)苯海拉明-d3和利多卡因-d10的混合内标溶液100μL,涡旋混匀后静置30 min,用70%(体积分数)乙腈溶液稀释至5.00 mL。涡旋混合2 min,加入乙腈饱和的正己烷溶液1.0 mL和氯化钠饱和溶液1.0 mL,超声20 min,涡旋15 min,于-20℃放置1 h。弃去上层正己烷相,取中间层相,加入等体积水稀释,离心5 min。取上清液过0.22μm滤膜,滤液按照超高效液相色谱-串联质谱法(UHPLC-MS/MS)测定,内标法定量。在色谱分析中,以Waters Acquity UPLC^(■)BEH C_(18)色谱柱为固定相,不同体积比的0.1%(体积分数)甲酸溶液和甲醇的混合溶液为流动相进行梯度洗脱分离。在质谱分析时,以电喷雾离子源正离子模式电离,在多反应监测模式下检测。结果显示:苯海拉明和利多卡因的质量浓度在0.50~40.0μg·L^(-1)内与目标组分和内标峰面积比值呈线性关系,检出限(3S/N)分别为8.12,3.27μg·kg^(-1);按照标准加入法进行回收试验,回收率为88.5%~105%;对加标阴性样品进行日内、日间精密度试验,峰面积比值的相对标准偏差(n=6)均小于6.0%;方法用于市售84份抗(抑)菌类消毒产品的分析,苯海拉明和利多卡因的总检出率为4.76%,其中1份样品中苯海拉明的检出量为2.86×10^(6)μg·kg^(-1)。

An aliquot(0.200 g)of antifungal disinfection product sample was taken,and mixed with 100μL of 1.00 mg L^(-1) mixed internal standard solution of diphenhydramine-ds and lidocaine-dio by vortex.The mixture was settled for 30 min,and diluted to 5.00 mL with 70%(volume fraction)acetonitrile solution.The solution obtained was mixed by vortex for 2 min,and 1.0 mL of n-hexane solution saturated with acetonitrile and 1.0 mL of sodium chloride saturated solution were added.After ultrasonicating for 20 min,and vertexing for 15 min.the mixed solution was stored at-2o℃C for 1 h.The upper n-hexane phase was discarded,and the middle layer phase was collected,and diluted by equal volume of water.After centrifuging for 5 min,the supernatant was taken and passed through a O.22μm filter membrane.The filtrate was determined by ultra-high performance liquid chromatography tandem mass spectrometry(UHPLC-MS/MS),and quantified by internal standard method.In chromatographic analysis,Waters Equity UPLC0^(6) BEH C_(18) column was used as the stationary phase,and gradient elution separation was performed with the mixtures of o.1%(volume fraction)formic acid solution and methanol solution at different volume ratios as the mobile phase.In mass spectrometry analysis,the positive ion mode of the electrospray ion source was used for ionization,and the multiple reaction monitoring mode was used for detection.It was shown that linear relationships between values of the mass concentration and the peak area ratio of the target component to the internal standard of diphenhydramine and lidocaine were kept in the range of 0.50-40.0μg·L^(-1),with detection limits(3S/N)of 8.12,3.27μg·kg^(-1),respectively.Test for recovery was made by the standard addition method,giving results in the range of 88.5%-105%.Tests for intra-day and inter-day precision were performed on spiked negative samples,with RSDs(n=6)of the peak area ratios less than 6.0%.The proposed method was applied for the analysis of 84 commercially available antifungal disinfection products.The total detection rate of diphenhydramine and lidocaine was 4.76%,of which the detection amount of diphenhydramine in one sample was 2.86X10^(6)μg·kg^(-1).