德拉马尼血药浓度测定方法的建立及应用

Establishment and application of a method for the determination of plasma concentration of delamanid

目的建立测定德拉马尼(DLM)血药浓度的方法并应用。方法血浆样品经甲醇沉淀蛋白后,采用超高效液相色谱-串联质谱(LC-MS/MS)法检测。色谱柱为Phenomenex Synergi^(TM) Fusion-RP,流动相为甲醇-0.1%甲酸溶液(梯度洗脱),柱温为40℃,流速为0.3 mL/min,进样量为1μL;离子源为电喷雾离子源,以多反应监测模式进行正离子扫描,用于定量分析的DLM离子对为m/z 535.0→352.0。采用上述LC-MS/MS法测定6例耐多药结核病(MDR-TB)患者体内DLM的血药浓度。结果DLM检测质量浓度的线性范围为0.05~8μg/mL(r=0.9995),定量下限为0.05μg/mL;日内、日间精密度RSD均小于10%,准确度为92.7%~104.9%,平均基质效应为94.3%~107.5%,平均萃取回收率为93.2%~98.1%。6例MDR-TB患者体内DLM的血药浓度为0.61~2.76μg/mL,平均(1.67±0.74)μg/mL。结论所建LC-MS/MS法专属性好,灵敏度、准确度及精密度均高,可用于MDR-TB患者体内DLM血药浓度的检测。

OBJECTIVE To establish and apply a method for the determination of plasma concentration of delamanid(DLM).METHODS After plasma samples were treated with methanol precipitation,LC-MS/MS was adopted to determine the plasma concentration of DLM.The chromatographic column was Phenomenex Synergi^(TM) Fusion-RP with mobile phase of methanol-0.1%formic acid solution(gradient elution).The column temperature was 40℃,the flow rate was 0.3 mL/min,and the sample size was 1μL.The ion source was electrospray ion source,and positive ion scanning was carried out in multi-reaction monitoring mode.The DLM ion pair used for quantitative analysis was m/z 535.0→352.0.The plasma concentration of DLM in 6 multidrug resistant tuberculosis(MDR-TB)patients were determined by the LC-MS/MS method.RESULTS The linear range of DLM was 0.05-8μg/mL(r=0.9995),and the lowest limit of quantitation was 0.05μg/mL.RSDs of intra-batch and inter-batch precision were all less than 10%.The accuracy ranged 92.7%-104.9%.Average matrix effect was 94.3%-107.5%.Average recoveries were 93.2%-98.1%.The plasma concentration of DLM in 6 MDR-TB patients ranged from 0.61-2.76μg/mL,with an average of(1.67±0.74)μg/mL.CONCLUSIONS The established LC-MS/MS analysis method has good specificity,high sensitivity,accuracy and precision,and can be used to determine DLM plasma concentration in MDR-TB patients.