A simple, sensitive and accurate LC-MS/MS method was developed and validated for the simultaneous quantification of apixaban (APB) and metformin (MET) in rat plasma using rivaroxaban as internal standard (IS). An Inertsil ODS3 C18 column (150 × 4.6 mm, 5 μm) was used for chromatographic separation with isocratic elution. Multiple reaction monitoring (MRM) using positive-ion ESI mode to monitor ion transitions of m/z 459.8 → 442.8 for APB, m/z 130.2 → 71.2 for MET, m/z 436.8 → 144.9 for IS. The procedure of method validation included selectivity, linearity, precision, accuracy, matrix effect, extraction recovery and stability were conducted according to the guidelines of EMA and FDA. The method was validated over the concentration range of 0.5-250 ng/mL for APB and 8-8000 ng/mL for MET. The intra- and inter-day precision and accuracy of the quality control samples exhibited relative standard deviations (RSD) < 12.5% and the accuracy values ranged from -8.6 to 12.4%. Recovery and matrix effect values variations were all less than 15%. After oral administration APB and MET to rats, the comparison of pharmacokinetic parameters of APB in the single and co-administrated groups showed significant difference in AUC(0-t) from 730.71 ± 121.31 to 573.07 ± 90.13 ng/mL·h, t1/2 from 5.86 ± 3.21 to 4.24 ± 1.15 h and Cmax from113.54 ± 24.04 to 159.42 ± 54.6 ng/mL. The comparison of pharmacokinetic parameters of MET in the single and co-administrated groups showed significant difference in t1/2 from 2.83 ± 1.81 to 3.97 ± 0.57 h and Cmax from 4015.76 ± 873.23 to 3153.6 ± 1012.51 ng/mL. The results indicated that drug-drug interactions (DDI) occurred might be owing to APB affect one or all of OCTs, MATE1, MATE2-K.

中文翻译：

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同时测定大鼠血浆中阿哌沙班和二甲双胍的 LC-MS/MS 方法测定：药代动力学药物-药物相互作用研究的评估。

开发并验证了一种简单、灵敏且准确的 LC-MS/MS 方法，可使用利伐沙班作为内标 (IS) 同时定量大鼠血浆中的阿哌沙班 (APB) 和二甲双胍 (MET)。使用 Inertsil ODS3 C18 柱（150 × 4.6 mm，5 μm）进行等度洗脱的色谱分离。使用正离子 ESI 模式进行多反应监测 (MRM)，监测 APB 的 m/z 459.8 → 442.8、MET 的 m/z 130.2 → 71.2、IS 的 m/z 436.8 → 144.9 的离子跃迁。方法验证程序包括选择性、线性、精密度、准确度、基质效应、提取回收率和稳定性均按照EMA和FDA的指南进行。该方法在 APB 0.5-250 ng/mL 和 MET 8-8000 ng/mL 浓度范围内进行了验证。质量控制样品的日内和日间精密度和准确度表现出相对标准偏差 (RSD) < 12.5%，准确度值范围为 -8.6 至 12.4%。回收率和基质效应值变化均小于 15%。大鼠口服APB和MET后，单用组和联合给药组APB药代动力学参数比较，AUC(0-t)从730.71±121.31升至573.07±90.13ng/mL·h，t1/ 2从5.86±3.21到4.24±1.15小时，Cmax从113.54±24.04到159.42±54.6ng/mL。单药组和联合给药组MET药代动力学参数比较显示，t1/2从2.83±1.81到3.97±0.57 h，Cmax从4015.76±873.23到3153.6±1012.51 ng/mL，差异有统计学意义。