气相色谱法测定胃蛋白酶中丙酮和异丙叉丙酮残留量

Determination of Acetone and Mesityl Oxide Residues in Pepsin by GC

目的建立测定胃蛋白酶原料药中丙酮和异丙叉丙酮溶剂残留量的气相色谱法。方法以纯化水为溶剂制备样品,85℃条件下顶空加热30 min;色谱柱为Agilent DB-Wax毛细管柱(30 m×0.25 mm,0.25µm),程序升温,检测器为火焰离子化检测器,氮气流量为1.0 mL/min,采用外标法定量。结果丙酮、异丙叉丙酮的质量浓度分别在50~400µg/mL(r=0.9994)和2~16µg/mL(r=0.9990)范围内与峰面积线性关系良好;丙酮和异丙叉丙酮的检测限分别为0.03,0.09µg/mL,定量限分别为0.10,0.30µg/mL;精密度试验中,丙酮、异丙叉丙酮峰面积的RSD分别为1.85%和2.16%(n=6);平均加样回收率分别为95.87%和95.00%,RSD分别为1.24%和1.90%(n=9)。结论该方法操作简便、结果准确、残留溶剂分离度好、灵敏度高、重复性良好,可用于胃蛋白酶原料药中有机溶剂残留量的检测。

Objective To establish the gas chromatography(GC) method for the determination of acetone and mesityl oxide residues in active pharmaceutical ingredients(APIs) of pepsin.Methods The samples were prepared with purified water as the solvent and heated at 85 ℃ in headspace for 30 min. The chromatographic column was Agilent DB-Wax capillary column(30 m × 0. 25 mm,0. 25 μm),the temperature was programmed,the detector was flame ionization detector,the flow rate of nitrogen was 1. 0 mL/min,and the external standard method was used for quantitative analysis.Results The linear ranges of acetone and mesityl oxide were50-400 μg/mL(r = 0. 999 4) and 2-16 μg/mL(r = 0. 999 0) respectively. The limits of detection(LOD) of acetone and mesityl oxide were 0. 03 μg/mL and 0. 09 μg/mL respectively,the limits of quantitation(LOQ) of acetone and mesityl oxide were 0. 10 μg/mL and 0. 30 μg/mL respectively. In the precision test,the RSDs of peak areas of acetone and mesityl oxide were 1. 85% and 2. 16%(n = 6) respectively. The average recoveries of acetone and mesityl oxide were 95. 87% and95. 00% with RSDs of 1. 24% and 1. 90%(n = 9) respectively.Conclusion and repeatable,which can be used for the determination of organic solvent residues in pepsin APIs.