【摘要】 目的:建立以液相色谱-质谱联用(LC/MS)法测定血清中甲氨蝶呤(MTX)的方法并用于临床适时监测,进一步探讨MTX剂量-血药浓度与药物不良反应(ADR)发生的相关性。方法:建立以ESI离子源,SCI(+)扫描方式,离解电压1.1 KV,雾化氮气1.5 L·min-1,干燥氮气15 L·min-1,干燥温度200℃,模块加热温度400℃,加氢目标离子m/z455,碎片离子455,456的LC/MS法测定血清中MTX的方法并进行方法学验证,对59例临床ALL患儿接受大剂量甲氨蝶呤(HD-MTX)化疗后48,72,96 h时间点271例次MTX血药浓度监测和ADR发生情况观察。结果:方法的回归方程Y=(6.139 28E7)X-528120(n=6,r=0.999 7),MTX血清浓度在0.01～160μmol·L-1范围线性良好,最低定量限0.01μmol·L-1(S/N≥3),通过方法学验证符合临床生物样本分析要求;对HD-MTX患儿化疗后48 h与72 h点血药浓度监测。结果组内两监测点血药浓度变异较大(CV%>97%),经独立样本t检验存在显著性差异(P=0.000),然而不同病理诊断分组间两时间点血药浓度无显著性差异(P>0.05);MTX剂量与ADR发生经多元变量非参数的偏相关分析结果相关系数rs=0.133。结论:该检测方法具有快捷、准确、痕量测定血中MTX浓度的特点,可用于临床及科研MTX血药浓度的监测;四氢叶酸钙(CF)的救援应通过监测MTX血药浓度并根据结果调整剂量,监测C48h和C72h点用于预判MTX蓄积中毒并指导CF救援有一定的临床实际意义;通过MTX监测血药浓度制定个体治疗方案,ADR发生与MTX剂量之间并非强相关。更多还原

【Abstract】 OBJECTIVE To establish a method for determination of the concentration of MTX by LC/MS and used in clinical monitor,to further explore the MTX dose-blood drug concentration and ADR correlation.METHODS The method was established and validated for determination of the concentration of serum MTX by LC/MS,under the condition of the ESI ion source,SCI(+) scanning,dissociation voltage 1.1 KV,atomizing nitrogen 1.5 L·min-1,dry nitrogen 15 L·min-1,drying temperature 200 ℃,module heating temperature 400 ℃,the M/Z is 455 target ion of hydrogenation,fragment ions of target 455,456,to determine the serum MTX.59 Patients with acute lymphoblastic leukemia(ALL) underwent chemotherapy with HD-MTX,271 cases of serum drug concentration were monitored at 48,72,and 96 h time point and the ADR was under observation.RESULTS The regression equation of method was Y=(6.139 28E7)X-528 120(n=6,r=0.999 7),there was good linearity when the range of serum concentration of MTX in 0.01 to 160 μ mol·L-1,the least limit of quantification was 0.01 μ mol·L-1(S/N≥3);it could meet the clinical requirements of biological samples analysis by the method validation.The coefficient of variation was larger(CV%>97%) through the comparison of the blood concentration monitoring results within a group chemotherapy after 48 h and 72 h.There were significant differences through the t-test of independent samples(P=0.000).However there was no significant difference in different pathological diagnostic subgroup two time-point(P>0.05),the correlation coefficient rs of the dose of MTX with ADR occurring was 0.133 via multivariate nonparametric correlation analysis.CONCLUSION The detection method has the features of quick,accurate,trace determination,can be used for clinical and scientific research.The dosage of CF rescue can be adjusted according to the results of the MTX blood concentration monitoring.Monitoring point of C48h and C72h has certain clinical significance for predicting MTX poisoning and as guidance of CF rescue.ADR and dose of MTX are not strong correlation when individualized treatment programs is designated by blood drug concentration.更多还原