二维高效液相色谱法测定血浆中氨甲环酸血药浓度方法的建立及其在关节置换术患者中的应用

目的：建立全自动二维色谱法（2D-LC-UV）测定血浆中氨甲环酸浓度的方法，用于临床上行关节置换术患者围术期氨甲环酸治疗药物浓度的监测。方法：待测物在一维柱ASTON SPX（100 mm×4.6 mm，5 μm）上初步分离，通过中间柱ASTO N SHC（10 mm×4.6 mm，3 μm）截取保留并在二维柱ASTON SX1（150 mm×4.6 mm，5 μm）上进一步分离，200 μL样品进样，并最终测定。流速均为1.0 mL·min^-1，柱温为40℃，紫外检测波长为220 nm。所建立方法运用于58名行膝关节/髋关节置换术的患者围术期中氨甲环酸血浆样本的检测分析。结果：在所建立的色谱条件下，氨甲环酸与各杂质分离良好，线性范围为5~300 μg·mL^-1，最低检测限为5 μg·mL^-1，批间批间精密度均小于6.6%，提取回收率大于72.4%。58名患者围术期中氨甲环酸首次给药后3个时间点（5 min，30 min和2 h）平均血浆浓度分别为：（103.3±20.4）μg·mL^-1，（53.0±14.8）μg·mL^-1和（22.8±8.3）μg·mL^-1。手术中男女之间氨甲环酸血药浓度无显著性差异（P>0.05），而不同手术种类之间（膝关节/髋关节置换术）氨甲环酸血药浓度存在显著性差异（P<0.05）。结论：此法操作简单，准确、精密度好，适用于临床氨甲环酸血浆浓度的监测及浓度-疗效关系研究。

Determination of tranexamic acid in human plasma by two-dimensional high performance liquid chromatography and its application in patients with joint arthroplasty

OBJECTIVE Tranexamic acid was quantified by a two-dimensional liquid chromatography coupled with ultraviolet detector (2D-LC-UV), for monitoring concentration of tranexamic acid in patients with joint arthroplasty.METHODS Tested samples were separated initially on a one-dimensional column ASTON SPX (100 mm×4.6 mm, 5 μm), and intercepted and retained by a middle column ASTO N SHC (10 mm×4.6 mm, 3 μm) and further separated on a two-dimensional column ASTON SX1 (150 mm×4.6 mm, 5 μm), 200 μL of sample was injected and measured finally. Flow rates were both at 1.0 mL·min^-1, and column temperature was 40℃, and UV wavelength of detection was 220 nm. Plasma samples of 58 patients with joint arthroplasty (including hip joint arthroplasty and knee joint arthroplasty) receiving tranexamic acid by intravenous route were collected and analyzed.RESULTS Under established chromatographic conditions, tranexamic acid was separated well with other impurities, and linear range was 5-300 μg·mL^-1, with the lowest detection limit of 5 μg·mL^-1. RSD of intra- and inter-day precision and accuracy for tranexamic acid was 6.6% or less. Average recovery of tranexamic acid was above 72.4%. Mean concentrations in plasma of 58 patients were (103.3±20.4) μg·mL^-1, (53.0±14.8) μg·mL^-1 and (22.8±8.3) μg·mL^-1, respectively at three sampling time points (5 min, 30 min and 2 h after administration). Significant difference was not observed between males and females (P>0.05), but between different kinds of joint arthroplasty (P<0.05). CONCLUSION This method is simple and precise, and can be used as an effective approach in clinic and research fields of tranexamic acid.