UPLC-MS/MS法检测唾液中的奥沙西泮
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摘要
目的建立唾液中奥沙西泮的超高液相色谱—串联质谱(UPLC-MS/MS)检测方法。方法液相色谱仪采用流动相为乙腈-0.1%甲酸水溶液进行梯度方式洗脱。串联质谱仪采用电喷雾离子源正离子(ESI+)多反应监测(MRM)模式进行质谱分析,用287.0/241.0、287.0/165.0和287.0/104.0这3对特征离子对和保留时间对奥沙西泮进行定性分析,采用外标标准曲线法对奥沙西泮进行定量分析。唾液样品经甲醇稀释后,经振荡器振荡3 min,在15 000 r·min~(-1)转速下离心3 min,取上清液过有机滤膜(13 mm×0.22μm)后待检。结果奥沙西泮在1~100 ng·mL~(-1)线性范围内,线性方程:A=3 700×C+2 390,线性关系良好(R2=0.9 999),检出限和定量限分别为0.09 ng·mL~(-1)和0.30 ng·mL~(-1);奥沙西泮的提取回收率均高于90%,日内及日间精密度小于10%。结论本方法简便、快速、定量准确,适用于检测唾液中的奥沙西泮。
Objective A method of determination of oxazepam in saliva was established by UPLC-MS/MS method. Methods The ultra high performance liquid chromatograph was using the mobile phase contisted of ACN( eluent A) and 0. 1% FA in water( eluent B) which was running in gradient elution way.The mass spectrometric acquisition was carried out by electospray ionization in positive mode( ESI+) with multiple reaction monitoring( MRM) method,multiple characteristic ion pairs( 287. 0/241. 0,287. 0/165. 0 and 287. 0/104. 0) and retention time were adopted to oxazepam for qualitative analysis,and the external standard method was adopted to oxazepam for quantitative analysis. The samples were diluted with methanol reagent,and the samples diluted were oscillated for 3 min,centrifuged for 3 min at 15 000 r · min~(-1),the supernatant liquor was to be detected after purificated by organic membrane filter. Results The linear ranges were 1 ～ 100 ng · mL~(-1) for oxazepam,the linear equation: A = 3 700 × C + 2 390,and the linear relationship was good with the correlation coefficient was 0. 9 999,the qualitative and quantitative restriction were 0. 09 ng·mL~(-1) and 0. 30 ng·mL~(-1) respectively in the saliva. The extract recovery of oxazepam was above 90%,intra-and inter-day precisions were all less than 10%. Conclusion This method is simple、rapid and accurate for determination of oxazepam in saliva.
Objective A method of determination of oxazepam in saliva was established by UPLC-MS/MS method. Methods The ultra high performance liquid chromatograph was using the mobile phase contisted of ACN( eluent A) and 0. 1% FA in water( eluent B) which was running in gradient elution way.The mass spectrometric acquisition was carried out by electospray ionization in positive mode( ESI+) with multiple reaction monitoring( MRM) method,multiple characteristic ion pairs( 287. 0/241. 0,287. 0/165. 0 and 287. 0/104. 0) and retention time were adopted to oxazepam for qualitative analysis,and the external standard method was adopted to oxazepam for quantitative analysis. The samples were diluted with methanol reagent,and the samples diluted were oscillated for 3 min,centrifuged for 3 min at 15 000 r · min~(-1),the supernatant liquor was to be detected after purificated by organic membrane filter. Results The linear ranges were 1 ～ 100 ng · mL~(-1) for oxazepam,the linear equation: A = 3 700 × C + 2 390,and the linear relationship was good with the correlation coefficient was 0. 9 999,the qualitative and quantitative restriction were 0. 09 ng·mL~(-1) and 0. 30 ng·mL~(-1) respectively in the saliva. The extract recovery of oxazepam was above 90%,intra-and inter-day precisions were all less than 10%. Conclusion This method is simple、rapid and accurate for determination of oxazepam in saliva.
引文
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[11]王伟,刘永锁,乔湜,等.UPLC-MS/MS法同时测定人血浆中卡马西平、拉莫三嗪、氯硝西泮、地西泮及其代谢物奥沙西泮浓度[J].中国现代应用药学,2013,30(11):1215-1219.
[12]Sauve EN,Langodegard M,Ekeberg D,et al.Determination of benzodiazepines in ante-mortem and post-mortem whole blood by solid-supported liquid-liquid extraction and UPUPLC-MS/MS[J].J Chromatogr B Analyt Technol Biomed Life Sci,2011,883(884):177-188.
[13]Verplaetse R1,Cuypers E,Tytgat J.The evaluation of the applicability of a high p H mobile phase in ultrahigh performance liquid chromatography tandem mass spectrometry analysis of benzodiazepines and benzodiazepine-like hypnotics in urine and biood[J].J Chromatogr A,2012,12(49):147-154.
[2]Simonsen KW,Hermansson S,Steentoft A,et al.A validated method for simultaneous screening and quantification of twentythree benzodiazepines and metabolites plus zopiclone and zaleplone in whole blood by liquid-liquid extraction and ultra-performance liquid chromatography-tandem mass spectrometry[J].J Anal Toxicol,2010,34(6):332-341.
[3]Goldberger BA,Chronister CW,Merves ML.Quantitation of benzodiazepines in blood and urine using gas chromatographymass spectrometry(GC-MS)[J].Methods Mol Biol,2010,603:75-87.
[4]朱小红,李涛,李建荣.苯二氮类药物的气相色谱—质谱联用法定性分析方法研究[J].药物分析杂志,2011,31(12):2289-2293.
[5]张韬,凌义.正相液相色谱法测定人体血浆中奥沙西泮的浓度[J].中国药物警戒,2012,9(5):257-259.
[6]郑付春,江泽斌.固相萃取—液相色谱法测定血浆中4种苯二氮类药物[J].汕头大学医学院学报,2014,27(4):208-210.
[7]徐晖.高液相色谱法同时测定苯巴比妥、卡马西平、艾司唑仑、阿普唑仑、氯硝西泮、奥沙西泮的血药浓度[J].海峡药学,2013,25(9):170-173.
[8]刘立新,崔一民.RP-HPLC同时测定人体外血浆中7种苯二氮类镇静催眠药的浓度[J].药物分析杂志,2011,31(2):336-339.
[9]Marin SJ,Mc Millin GA.LC-MS/MS analysis of 13 benzodiazepines and metabolites in urine,serum,plasma,and meconium[J].Methods Mol Biol,2010,603:89-105.
[10]李存,皇甫伟国,杨挺,等.液相色谱串联质谱法测定猪尿中10种镇静剂类药物残留量[J].分析化学研究简报,2010,38(7):1015-1018.
[11]王伟,刘永锁,乔湜,等.UPLC-MS/MS法同时测定人血浆中卡马西平、拉莫三嗪、氯硝西泮、地西泮及其代谢物奥沙西泮浓度[J].中国现代应用药学,2013,30(11):1215-1219.
[12]Sauve EN,Langodegard M,Ekeberg D,et al.Determination of benzodiazepines in ante-mortem and post-mortem whole blood by solid-supported liquid-liquid extraction and UPUPLC-MS/MS[J].J Chromatogr B Analyt Technol Biomed Life Sci,2011,883(884):177-188.
[13]Verplaetse R1,Cuypers E,Tytgat J.The evaluation of the applicability of a high p H mobile phase in ultrahigh performance liquid chromatography tandem mass spectrometry analysis of benzodiazepines and benzodiazepine-like hypnotics in urine and biood[J].J Chromatogr A,2012,12(49):147-154.