目的: 建立马来酸噻吗洛尔滴眼液对映异构体含量的HPLC测定方法。方法: 应用手性液相色谱法,采用InfinityLabPoroshell 120 Chiral-T柱色谱柱(100 mm×4.6 mm,2.7 μm),对左旋噻吗洛尔进行定量分析,以甲酸铵溶液(取甲酸铵1.26 g,加水900 mL使溶解,甲酸调pH至3.5,加水至1 000 mL)-无水乙醇(8∶92)为流动相,流速为0.5 mL·min-1;柱温为30 ℃;检测波长为297 nm。结果: 在此色谱条件下,测定的2批原料所制的6批样品含量分别为1.1%、1.1%、1.2%、0.053%、0.053%及0.057%;噻吗洛尔峰与对映异构体峰分离度良好(R＞2.0),噻吗洛尔在质量浓度0.5~15.3 μg·mL-1范围内呈良好的线性关系(r=1.000),对映异构体在质量浓度0.7~15.4 μg·mL-1范围内呈良好的线性关系(r=0.999 7),噻吗洛尔、对映异构体的定量限分别为0.5和0.6 μg·mL-1;12份供试品对映异构体含量均值为0.052%。结论: 本法准确可靠,可为马来酸噻吗洛尔滴眼液质量控制及我国药典相关标准的建立提供参考。
Objective: To establish an HPLC method for the determination of enantiomeric impurity of timolol in eye drops. Methods: Levotimolol was assayed by chiral liquid chromatography. Analysis was performed on an Infinity Lab Poroshell 120 Chiral-T (100 mm×4.6 mm, 2.7 μm) column. The mobile phase was ammonium formate buffer solution(dissolve 1.26 g ammonium formate in 900 mL water, adjust to pH 3.5 with formic acid and dilute to 1 000 mL with water)-anhydrous ethanol (8∶92). The flow rate was 0.5 mL·min-1 and the column temperature was 30 ℃. The detection wavelength was 297 nm. Results: Under the chromatographic conditions, the contents of six batches of samplesmade from two batches of raw materials were 1.1%, 1.1%, 1.2%, 0.053%, 0.053% and 0.057%, respectively. S- and R-timolol could be separated well(R＞2.0). The ranges of S- and R-timolol were 0.5-15.3 μg·mL-1(r=1.000) and 0.7-15.4 μg·mL-1showed good linear relationships, respectively. The LOQ of S-timolol was 0.5 μg·mL-1 and R-timolol was 0.6 μg·mL-1, respectively. The average content of enantiomeric impurity in 12 samples was 0.052%. Conclusion: The established method is accurate and reliable that can provide reference for the quality control of timolol maleate in eye drops and the establishment of relevant standards in Chinese Pharmacopoeia criteria.
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