This study aimed to establish a highly accurate method for quantifying methotrexate(MTX)concentrations in serum using an ultra-high performance liquid chromatography-tandem mass spectrometry system(UPLC-MS/MS)and to c...This study aimed to establish a highly accurate method for quantifying methotrexate(MTX)concentrations in serum using an ultra-high performance liquid chromatography-tandem mass spectrometry system(UPLC-MS/MS)and to compare its performance with the chemiluminescence microparticle immunoassay(CMIA).A total of 211 serum samples from pediatric patients with intracranial tumors undergoing high-dose MTX treatment were analyzed using both techniques.Correlation and agreement analyses were performed to assess the level of concordance between these methods.The results demonstrated a significant correlation(P<0.05)between the two methods,with correlation coefficients of 0.9913 and 0.9893,respectively.However,a statistical difference was noted in MTX serum concentrations at lower levels(0.04-1.5μmol/L),while no significant difference was observed at higher concentrations(1.5-400μmol/L).Specifically,in the 0.04-1.5μmol/L range(107 cases),Bland-Altman analysis indicated a bias of 0.018 between the two methods.Given the observed discrepancies,particularly at lower concentrations,it is advised that the UPLC-MS/MS method should not be used interchangeably with the CMIA assay for therapeutic drug monitoring in patients receiving high-dose MTX treatment.展开更多
建立柱前衍生化-UPLC-MS/MS方法检测达格列净原料药中的潜在的基因毒性杂质5-溴-2-氯苯甲酰氯。以苯胺作为衍生化试剂,采用Acouity UPLC Bed C18柱(50 mm×2.1 mm, 1.7μm)为色谱柱,0.1%甲酸-水体系作为流动相进行梯度洗脱,体积流量...建立柱前衍生化-UPLC-MS/MS方法检测达格列净原料药中的潜在的基因毒性杂质5-溴-2-氯苯甲酰氯。以苯胺作为衍生化试剂,采用Acouity UPLC Bed C18柱(50 mm×2.1 mm, 1.7μm)为色谱柱,0.1%甲酸-水体系作为流动相进行梯度洗脱,体积流量0.3 mL/min,进样量2μL,柱温25℃;在电喷雾离子源、正离子监测下,采用多反应扫描(MRM)模式进行检测。该方法的专属性强;在1~100 ng/mL范围内线性关系良好(r=0.999 3);检测限和定量限分别为0.05 ng/mL和0.10 ng/mL;低、中、高浓度(n=3)的平均加样回收率在95.1%~106.5%之间,RSD均小于3.0%;衍生化产物在室温下12 h内稳定。该方法操作简便,灵敏度高,适用于检测达格列净原料药中5-溴-2-氯苯甲酰氯。展开更多
基金The Science and Technology Fund of Beijing Shijitan Hospital(Grant No.2022-C06)the National Key Research and Development Program of China(Grant No.2020YFF01014606).
文摘This study aimed to establish a highly accurate method for quantifying methotrexate(MTX)concentrations in serum using an ultra-high performance liquid chromatography-tandem mass spectrometry system(UPLC-MS/MS)and to compare its performance with the chemiluminescence microparticle immunoassay(CMIA).A total of 211 serum samples from pediatric patients with intracranial tumors undergoing high-dose MTX treatment were analyzed using both techniques.Correlation and agreement analyses were performed to assess the level of concordance between these methods.The results demonstrated a significant correlation(P<0.05)between the two methods,with correlation coefficients of 0.9913 and 0.9893,respectively.However,a statistical difference was noted in MTX serum concentrations at lower levels(0.04-1.5μmol/L),while no significant difference was observed at higher concentrations(1.5-400μmol/L).Specifically,in the 0.04-1.5μmol/L range(107 cases),Bland-Altman analysis indicated a bias of 0.018 between the two methods.Given the observed discrepancies,particularly at lower concentrations,it is advised that the UPLC-MS/MS method should not be used interchangeably with the CMIA assay for therapeutic drug monitoring in patients receiving high-dose MTX treatment.
