Analytical Development, Shasun Research Center, 27 Vandalur Kelambakkam Road, Keelakottaiyur, Chennai, India.
J Pharm Biomed Anal. 2011 Mar 25;54(4):850-4. doi: 10.1016/j.jpba.2010.10.015. Epub 2010 Oct 28.
A new chiral purity method was developed for D-cycloserine (D-cys) by reverse phase HPLC and validated. Chiral derivatizing reagents, viz., o-phthalaldehyde and N-acetyl-L-cysteine were utilized in this method. The resultant diastereomers were resolved using Zorbax SB Phenyl HPLC column under isocratic elution. A mobile phase of 95:05 (v/v), 20mM Na(2)HPO(4) (pH 7), and acetonitrile, respectively, was used with the flow rate of 1.0 mL/min and UV detection at 335 nm. The method development with different chiral stationary phases and chiral derivatization reagents were also investigated. The stability of diastereomer derivative and influence of organic modifier and pH of the mobile phase were studied and optimized. The stability-indicating capability of the method was established by performing stress studies under acidic, basic, oxidation, light, humidity and thermal conditions. The detection and quantitation limit of L-cycloserine (L-cys) were 0.015 and 0.05% (w/w), respectively. A linear range from 0.05 to 0.30% (w/w) was obtained with the coefficient of determination (r(2)) 0.998. The recovery obtained for L-cys was between 92.9 and 100.2%. This method was applied successfully in pharmaceutical analysis to determine the content of L-cys in D-cys bulk drug.
建立了一种新的手性纯度方法，用于 D-环丝氨酸（D-cys）的反相高效液相色谱法和验证。手性衍生试剂，即邻苯二醛和 N-乙酰-L-半胱氨酸，用于该方法。使用 Zorbax SB Phenyl HPLC 柱在等度洗脱下分离所得非对映异构体。流动相为 95:05（v/v）、20mM Na 2 HPO 4 （pH 7）和乙腈，流速为 1.0 mL/min，UV 检测波长为 335nm。还研究了不同的手性固定相和手性衍生试剂的方法开发。研究并优化了非对映异构体衍生物的稳定性以及流动相有机改性剂和 pH 的影响。通过在酸性、碱性、氧化、光照、湿度和热条件下进行稳定性研究，建立了方法的稳定性指示能力。L-环丝氨酸（L-cys）的检测限和定量限分别为 0.015 和 0.05%（w/w）。线性范围为 0.05-0.30%（w/w），相关系数（r 2 ）为 0.998。L-cys 的回收率在 92.9%至 100.2%之间。该方法成功应用于药物分析，用于测定 D-cys 原料药中 L-cys 的含量。
