超高效液相色谱-串联质谱法检验腐败血中吗啡和6-单乙酰吗啡

采用改良的QuEChERS前处理方法,建立了同时检验腐败血中吗啡和6-单乙酰吗啡的超高效液相色谱-串联质谱(UPLC-MS/MS)方法。腐败血中加入乙腈-水(4∶1,v/v)混合溶液、30 mg NaCl和60 mg MgSO4盐析促进相分离,最后经25 mg N-丙基乙二胺(PSA)和25 mg MgSO4净化。选用ZORBAX Eclipse Plus C18色谱柱分离,0. 01%(v/v)氨水和乙腈作为流动相进行梯度洗脱,电喷雾电离正离子(ESI+)模式扫描,多反应监测(MRM)模式检测。结果表明,吗啡和6-单乙酰吗啡在5～200μg/L范围内线性关系良好,相关系数(r2)≥0. 995 7,检出限(S/N=3)均为1μg/L,定量限(S/N=10)均为5μg/L。3个加标水平(5、100和200μg/L)下,吗啡和6-单乙酰吗啡的平均加标回收率分别为81. 84%～103. 44%和81. 03%～104. 46%,日内和日间精密度(RSD)均小于12%,基质效应为83. 04%～107. 61%。方法简便、灵敏、可靠,可实现腐败血中吗啡和6-单乙酰吗啡的快速定性定量检验。

Determination of morphine and 6-monoacetylmorphine in putrefied blood using ultra performance liquid chromatography-tandem mass spectrometry

This study was performed to develop an ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method to detect and quantify morphine and 6-monoacetylmorphine in putrefied blood for forensic toxicological purposes. A modified QuEChERS method was implemented as follows:extraction of morphine and 6-monoacetylmorphine from putrefied blood was performed with an acetonitrile-water (4:1, v/v) mixture, and 30 mg NaCl and 60 mg anhydrous MgSO₄ were subsequently added as salting out agents to induce phase separation. The supernatant was cleaned by adding 25 mg primary secondary amine sorbent (PSA) and 25 mg anhydrous MgSO₄. Separation of the target compounds was performed using a ZORBAX Eclipse Plus C₁₈ column by UPLC over a 4 min gradient elution where 0.01% (v/v) ammonium hydroxide buffer (A) and acetonitrile (B) were used as the mobile phase. MS/MS was used in positive electrospray ionization (ESI⁺) mode and multiple-reaction monitoring (MRM) was performed to detect the target drugs. This method achieved satisfactory recoveries (R) for morphine and 6-monoacetylmorphine with mean recovery values ranging from 81.84% to 103.44% and 81.03% to 104.46% respectively. The developed method also provided efficient purification of the sample from endogenous interferences with matrix effect (ME) ranging from 83.04% to 107.61%. The method was validated and the limit of detection (LOD) and limit of quantification (LOQ) were 1 and 5 μg/L, respectively, for both compounds and the precision with RSD ranged from 1% to 12%. This method proved to be quick, sensitive, rugged, and suitable for the analysis of illegal drugs in putrefied whole blood.