DETERMINATION OF COCAINE AND BENZOYLECGONINE IN HUMAN PLASMA BY LC-MS/MS
DETERMINATION OF COCAINE AND BENZOYLECGONINE IN HUMAN PLASMA BY LC-MS/MS
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采用 LC-MS/MS 测定人血浆中的可卡因和苯甲酰爱康宁
DOI：
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发表时间：
2009
期刊：
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通讯作者：
F. Loghin
中科院分区：
文献类型：
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作者：
D. Popa;L. Vlase;F. Loghin
建立了同时测定人血浆中可卡因(COC)和苯甲酰ecGonine(BZE)浓度的高效液相色谱/质谱联用(LC/MS/MS)方法。血浆样品经甲醇沉淀蛋白质后，在Zorbax SB-C18色谱柱上，在等度条件下，甲醇-0.1%甲酸-水的32：68(v/v)混合溶液在45oC下分离，流速为1mL/min。检测模式为多反应监测(m/z290→m/z168和m/z304→m/z182)。BZE和COC的标准曲线均在20-2000 ng/mL范围内(r>；0.992)。在没有配备LC/MS/MS系统的实验室中，对COC和BZE的LC分析需要先前的分离步骤。为此，本研究采用LC/MS/MS联用技术，研究了从人血浆中分离COC和BZE的最佳实验条件。在不同的pH值下对几种混合溶剂和固相萃取柱进行了测试。OASIS HLB柱上固相萃取的结果最好(COC和BZE的平均回收率分别为71%和92.4%，n=3)。所建立的LC/MS/MS方法和最佳提取条件均可成功应用于临床和法医毒理学快速测定人血浆中的COC和BZE。用ă快速ăLC/MS/MS法检测血浆ă中的苯偶氮卓西林(BZE)。ă/MS/MS检测血浆中的苯偶氮环丙氨酸(BZE)。DUPă沉淀区蛋白或铜偏醇在血浆ă，分析ţII S-Au分离在条件ăţII的Zorbax SBC18中的蛋白质或铜偏醇在条件ţII的浮游生物32：68(v/v)de metanol si soluţ，即载脂蛋白ă的酸甲酸0，1%，La 45oC铜借方1毫升/分钟。ţia S--一种在模块中的实现(m/z 290→m/z 168 pentu BZE，m/z 304→m/z 182 pentu COC)。CURBELE DE CALIBRARE是S-Au Realizat pe Domeiul de ConceraţII 202000 ng/mL atát pentu BZE cát si pentu COC(r>；0.992)。在实验室护理中，我们必须建立LC/MS/MS系统，分析LC和ăăde izolare。Astfel，Metoda LC/MS/MS详述了ăS--一种在ţ实验研究中的应用。操作时间的需要是izolăRII coc si bze in血浆ă。S说：“ţi si cartuse de Extrcţie pe fazăSolidă(EFS)la Diferite valori de pH.CELE MAI BUNE REZZUTATE S-Au obţiNut Prin EFS cu cartuse OASIS HLB(注册ăSiri Medii de 71%pentu COC si 92.4%pentu BZE)。在Metoda LC/MS/MS中详细阐述了ăcát si cele mai Bune metode de exexcţie evidenţse pot applica cu在毒物诊所ăsi MedicalLawăin vederea DozăRII Rapid a COC si Bze in血浆ă中获得成功。
A high-throughput Liquid chromatography/ Mass spectrometry/ Mass spectrometry (LC/MS/MS) method for simultaneous quantification of cocaine (COC) and benzoylecgonine (BZE) in human plasma was elaborated. After the protein precipitation with methanol in the plasma samples, the analytes were separated on a Zorbax SB-C18 column under isocratic conditions using a 32:68 (v/v) mixture of methanol and 0.1% formic acid in water at 45oC, with a flow rate of 1 mL/min. The detection was in multiple reaction monitoring (MRM) mode (m/z 290 → m/z 168 for BZE and m/z 304 → m/z 182 for COC, respectively). Calibration curves were generated over the range of 20-2000 ng/mL for both BZE and COC (r > 0.992). In the laboratories that are not equipped with a LC/MS/MS system, the LC analysis of COC and BZE requires a previous isolation step. Therefore, the elaborated LC/MS/MS method was applied for the study of optimum experimental conditions for the isolation of COC and BZE from human plasma. Several solvent mixtures and solid-phase extraction (SPE) cartridges were tested at different pH values. The best results for both analytes were obtained by SPE on Oasis HLB cartridges (mean recovery of 71 % for COC and 92.4 % for BZE, n = 3). Both the elaborated LC/MS/MS method and the best extraction conditions can successfully be applied in clinical and forensic toxicology for the rapid quantification of COC and BZE in human plasma. Rezumat S-a elaborat o metodă rapidă LC/MS/MS pentru cuantificarea simultană a cocainei (COC) si benzoilecgoninei (BZE) din plasma umană. După precipitarea proteinelor cu metanol in probele de plasmă, analiţii s-au separat pe o coloană Zorbax SBC18 in condiţii izocratice folosind un amestec 32:68 (v/v) de metanol si soluţie apoasă de acid formic 0,1%, la 45oC cu un debit de 1 mL/min. Detecţia s-a realizat in modul MRM (monitorizarea reacţiilor multiple) (m/z 290 → m/z 168 pentru BZE, respectiv m/z 304 → m/z 182 pentru COC). Curbele de calibrare s-au realizat pe domeniul de concentraţii 202000 ng/mL atât pentru BZE cât si pentru COC (r > 0.992). In laboratoarele care nu sunt echipate cu un sistem LC/MS/MS, analiza LC a COC si BZE necesită anterior o etapă de izolare. Astfel, metoda LC/MS/MS elaborată s-a aplicat in studiul condiţiilor experimentale optime necesare izolării COC si BZE din plasma umană. S-au testat mai multe amestecuri de solvenţi si cartuse de extracţie pe fază solidă (EFS) la diferite valori de pH. Cele mai bune rezultate s-au obţinut prin EFS cu cartuse Oasis HLB (regăsiri medii de 71 % pentru COC si 92.4 % pentru BZE). Atât metoda LC/MS/MS elaborată cât si cele mai bune metode de extracţie evidenţiate se pot aplica cu succes in toxicologia clinică si medicolegală in vederea dozării rapide a COC si BZE din plasma umană.