青霉胺对映体的毛细管电泳手性分离及应用研究

Enantioseparation of Penicillamine by Capillary Electrophoresis

建立了一种青霉胺对映体的毛细管电泳手性分离方法.采用2,4-二硝基氯苯作为青霉胺的衍生试剂,以磺化-β-环糊精（S-β-CD）作为手性选择剂,50 mmol/L pH 9.5的硼砂缓冲溶液作为电泳缓冲液,在14 min 内实现了青霉胺对映体的毛细管电泳手性拆分,分离度达3.7.本文还考察了在大量D-青霉胺存在下, 测定微量L-青霉胺的可能性.当D-青霉胺中,L-青霉胺的含量在0.3%～2.0%范围内时,其浓度与吸光度之间呈现良好的线性关系（r＝0.9995）, 对D-青霉胺药片进行光学纯度分析,获得了满意的结果.

A method is presented for the chiral separation of penicillamine enantiomers. The peniciUamine enantiomers were derivatized with 1-chloro-2,4-dinitrobenzene and separated by capillary electrophoresis. The β-cyclodextrin （β-CD）, methyl-cyclodextrin （M-β-CD）, hydroxypropyl-β-CD （H-β-CD） and sulfated-β-CD （ S-β-CD）, as chiral selector, were examinedfor the enantionmefic separation of penicillamine, the concentration of chiral selector, some other experimental factors have also been optimized, such as concentration of borate buffer, pH of electrolyte and applied voltage. Optimal separation was obtained at pH 9.5, 50 mmol/L borate solution containing 25 mmol/L S-β-CD, as well as 20 kV applied voltage and 20℃ capillary temperature. Determination was followed by ultraviolet absorptiometric measurements at 202 nm. The derivatives of penicillamine enantiomers were resolved completely in 14 minutes, giving a rcsolution reached up to 3.7. The linear range, the recovery and the repeatability of L-penicillamine in the presence of large D-penicillamine were tested and a satisfactory result was obtained. The developed method has been employed for optical purity analysis of D-penicillamine drug, and allowed the determination of 0.3% L-penicillamine in D-penicillamine with well peak identification.