超高效合相色谱法测定l-OTS的含量及光学纯度
网络出版日期: 2017-06-02
基金资助
高校教师专业发展计划(BJ13100150032);国家自然科学基金(81171245)
Determination of l-OTS level and optical purity by ultra performance convergence chromatography method
Online published: 2017-06-02
Supported by
Professional Development Program for University Teachers, BJ13100150032; National Natural Science Foundation of China, 81171245
目的 建立l-OTS的含量测定及光学纯度检查的超高效合相色谱法(UPC2)。方法 使用ACQUITY UPC2TM TrefoilTMAMY1手性色谱柱(3.0 mm×50 mm,2.5 μm),柱温30 ℃,以超临界CO2和0.2%二乙胺的异丙醇(95∶5,体积比)为流动相,流速为1.0 mL/min,检测波长227 nm(补偿参比为310~360 nm),采用对照品法进行定量测定。结果 l-OTS与其对映异构体(d-OTS)有效分离,分离度为2.30,在40 ~1 000 mg/L范围内峰面积对应质量浓度呈良好的线性关系(r=0.999 9, n=7),平均加样回收率为99.8%(n=9),样品在24 h内稳定(RSD=0.81%),最低检测限5 ng(S/N=4)。结论 此方法简便、迅速,经方法学验证可用于l-OTS的含量测定及光学纯度分析。
谢一凡 , 杨若林 , 刘慧中 , 等 . 超高效合相色谱法测定l-OTS的含量及光学纯度[J]. 上海交通大学学报(医学版), 2016 , 36(3) : 459 . DOI: 10.3969/j.issn.16748115.2016.03.031
Objective To develop an ultra performance convergence chromatography (UPC2) method for determining l-OTS level and optical purity. Methods Reference substance method was used for quantitative determination. The separation of enantiomers was achieved on a chiral chromatographic column ACQUITY UPC2TM TrefoilTMAMY1(3.0 mm×50 mm, 2.5 mm) at the column temperature of 30 ℃ with the mobile phase of supercritical carbon dioxide-0.2% DEA in 2-propanol (95∶5) at a flow rate of 1.0 mL/min. Detection wavelength was set at 227 nm (compensation reference was 310-360 nm). Results l-OTS and its enantiomer (d-OTS) were effectively separated with the resolution of 2.30. Peak area was linear to mass concentration within the concentration rang of 40-1 000 mg/L (r=0.999 9, n=7). Average recovery was 99.8% (n=9) and the sample was stable within 24 h (RSD=0.81%). The LOD was 5 ng (S/N=4). Conclusion This method is simple and quick and can be used for determining l-OTS level and analyzing optical purity, which has been validated by methodology.
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