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目的建立测定(S)-OTS及其水解物6β-羟基-3α-对甲苯磺酰氧基莨菪烷(HTT)和对甲苯磺酸(TsOH)含量的高效液相色谱(HPLC)法,考察抗青光眼新药(S)-OTS·HCl水溶液的稳定性。方法 HPLC色谱条件:Diamonsil-C18为色谱柱;甲醇-10mmol/L磷酸二氢钠缓冲液(含5mmol/L四丁基溴化铵)(53:47)为流动相;流速1.0mL/min,紫外检测波长227nm,柱温31℃,进样量10μL。对建立的HPLC法进行相关验证试验。以0.03%(药用浓度)的(S)-OTS·HCl水溶液作为供试样品,利用已建立的HPLC法检测其溶质含量以考察稳定性。结果专属性试验显示,(S)-OTS与HTT和TsOH的色谱峰分离良好。线性关系试验表明,在系列对照品标准浓度范围内[(S)-OTS·HCl6.066～606.600μg/mL,HTT5.304～530.400μg/mL,TsOH3.034～303.400μg/mL],对照品(S)-OTS·HCl、HTT和TsOH与相应色谱峰面积呈线性相关(r2>0.999);灵敏度(检测限和定量限)、精密度、稳定性和方法回收率试验均符合要求。供试样品在室温下水解速度较快,其主成分(S)-OTS含量下降与水解物TsOH含量上升相吻合。结论建立了对主成分(S)-OTS及其水解物HTT和TsOH进行定量分析的HPLC法。药用浓度的(S)-OTS·HCl水溶液的稳定性受环境温度的影响,其水解易发生在磺酸酯部位。
OBJECTIVE To establish a high performance liquid chromatography (HPLC) method for the determination of (S) -OTS and its hydrolyzate 6β-hydroxy-3α-p-toluenesulfonyloxyethane (HTT) and p-toluenesulfonic acid (TsOH) Glaucoma new drug (S) -OTS · HCl aqueous solution stability. Methods HPLC conditions: Diamonsil-C18 column; methanol-10mmol / L sodium dihydrogen phosphate buffer (containing 5mmol / L tetrabutylammonium bromide) (53:47) as the mobile phase; UV detection wavelength 227nm, column temperature 31 ℃, injection volume 10μL. The established HPLC method for the relevant validation test. The aqueous solution of (S) -OTS · HCl at 0.03% (medicinal concentration) was used as the test sample, and its solute content was determined by established HPLC method to investigate its stability. The results of a proprietary test showed that (S) -OTS was well separated from the peaks of HTT and TsOH. Linear relationship test showed that the standard reference range of the series of reference substance [(S) -OTS · HCl6.066 ~ 606.600μg / mL, HTT5.304 ~ 530.400μg / mL, TsOH3.034 ~ 303.400μg / mL] (S) -OTS · HCl, HTT and TsOH linearly correlated with the corresponding chromatographic peak area (r2> 0.999). The sensitivity (limit of detection and limit of quantification), precision, stability and method recovery were satisfactory. The test sample hydrolyzed faster at room temperature, the main component (S) -OTS content decreased with the hydrolysis of TsOH content is consistent. Conclusion The HPLC method was established for the quantitative analysis of main component (S) -OTS and its hydrolyzate HTT and TsOH. The stability of (S) -OTS · HCl aqueous solution at the medicinal concentration is affected by the temperature of the environment, and its hydrolysis easily occurs at the sulfonate ester site.