GC-MS法测定甲磺酸达比加群酯中的致突变杂质

目的 建立一种气相色谱-质谱联用法（GC-MS）同时测定甲磺酸达比加群酯中甲磺酸甲酯、甲磺酸乙酯和甲磺酸异丙酯的方法。方法 采用DB-624色谱柱（30 m×0.25 mm,1.4 μm）;程序升温：起始温度100 ℃,以15 ℃·min^-1的速率升温至200 ℃,再以25 ℃·min^-1的速率升温至240 ℃;进样口温度为220 ℃,载气为氦气,流速为1.0 mL·min^-1,进样量为2.0 μL。采集模式为选择离子监测（SIM）,离子源温度为230 ℃。通过内标法进行计算。结果 三种甲磺酸酯之间的分离度均大于2.0,甲磺酸甲酯、甲磺酸乙酯、甲磺酸异丙酯的线性回归方程分别为Y=0.0130X+0.0524（r=0.999 0）、Y=0.0249X+0.0633（r=0.999 3）、Y=0.0188X+0.0906（r=0.998 5）,均在5.0～120.0 ng·mL^-1浓度范围内线性关系良好。检出限为1.5 ng·mL^-1（0.15 ppm）,定量限为5 ng·mL^-1（0.5 ppm）。甲磺酸甲酯、甲磺酸乙酯和甲磺酸异丙酯的平均回收率分别为96.28%、97.91%和95.87%,RSD分别为2.83%、2.93%和1.73%。结论 本方法简便、快速、准确、专属性强、灵敏度高,适用于甲磺酸达比加群酯中三种甲磺酸酯的检测。     

The anticoagulant effect of therapeutic levels of dabigatran in atrial fibrillation evaluated by thrombelastography (TEG®), Hemoclot Thrombin Inhibitor (HTI) assay and Ecarin Clotting Time (ECT)

Monitoring the effect of dabigatran (Pradaxa^®) is challenging. The aim of this study was to evaluate if thrombelastography reaction time (TEG^® R) could detect the anticoagulant effect of dabigatran showing a correlation between TEG^® R, Hemoclot Thrombin Inhibitor (HTI) assay and Ecarin Clotting Time (ECT) in patients with non-valvular atrial fibrillation (NVAF). Blood samples from 35?AF patients receiving either 110?mg (n 19) or 150?mg (n 16) dabigatran twice daily were analyzed with TEG^®, HTI and ECT 2–3?h after dabigatran intake. All patients had prolonged TEG^® R. The patients receiving dabigatran 110?mg ×2 had a TEG^® R mean 14.2?min (range 9.1–25), a mean dabigatran concentration measured by HTI of 268.5?ng/mL (range 54–837?ng/mL) and by ECT of 355.7?ng/mL (range 40–1020?ng/mL). The corresponding numbers for patients receiving dabigatran 150?mg ×2 were TEG^® R mean of 12.5?min (range 9.2–23.2?min), mean dabigatran concentration of 179.2?ng/mL by HTI (range 26–687?ng/mL) and by ECT 225.1?ng/mL (range 42–1020?ng/mL). The two dosage groups had comparable anticoagulation demonstrated by equally prolonged TEG^® R (p?=?.909), HTI (p?=?.707) and ECT (p?=?.567). No difference in creatinine levels in the two dosage groups was observed (p?=?.204) though patients with dabigatran concentration >400?ng/mL had significantly higher creatinine levels (p?=?.001). Large individual variation of the anticoagulant response was observed. Some patients had TEG^® R values up to three times upper normal limit with immediate risk of bleeding. Our data indicate that TEG^® R reflected dabigatran levels in NVAF patients and that TEG^® R correlated to HTI and ECT.