手性固定相高效液相色谱法测定皮肤渗透液中普萘洛尔对映体的研究

Establishment of Stereospecific Assay of Propranolol Enantiomers by Chiral Stationary Phase HPLC Method and its Application to Transdermal Delivery

目的建立普萘洛尔(PL)对映体分离的手性固定相RP-HPLC方法,用于普萘洛尔经皮渗透的对映体选择性研究。方法采用直链淀粉衍生物手性固定相Chiralpak AD-RH(4.6 mm×150 mm,5μm)色谱柱,以10 mmol·L-1硼砂缓冲液(0.1%三乙胺,pH 9.0)-乙腈-乙醇(50∶25∶25)为流动相分离普萘洛尔对映体,流速0.5 mL·min-1,柱温25℃,紫外波长为229 nm。另以硼砂缓冲液为皮肤渗透供给液,研究普萘洛尔对映体及其外消旋体经离体大鼠皮肤渗透的对映体选择性。结果普萘洛尔对映体在21 min内完全分离(分离度Rf=2.2),在3～200μg·mL-1内线性关系良好,R-普萘洛尔和S-普萘洛尔的平均回收率分别为100.84%和103.24%。另皮肤渗透实验结果表明,普萘洛尔外消旋体与对映体、及对映体之间经离体大鼠皮肤的渗透速率无显著差异。结论所建立的手性固定相RP-HPLC法准确、灵敏、高效,成功应用于普萘洛尔经皮渗透的对映体选择性研究。

OBJECTIVE To develop an RP-HPLC method for chiral separation of propranolol （PL） enantiomers and to apply it in enantioselective transdermal permeation study of PL. METHODS The separation was performed on an amylose derivative chiral stationary phase Chiralpak AD-RH column （4. 6 mm x 150 mm,5 μm） with the mobile phase of 10 mmol · L-1 borate buffer （0. 1% triethylamine, pH 9. 0）-acetonitrile-ethanol （50： 25： 25, V/V/V）. The flow rate was 0. 5 mL· min-1 , the column temperature was 25 .C, and the UV detection wavelength was 229 nm. Additionally, the enantioselectivity of racemie PL and enantiomers through excised mouse skin was evaluated by using borate buffer as permeation donor solvent. RESULTS PL enantiomers were entirely separated within 21 min （ Rf =2. 2） and were found to be linear in the range of 3 - 200 μg ·mL-1. The average recoveries of R-PL and S-PL were 100. 84% and 103.24% , respectively. The results of skin permeation experiments indicated that there were no significant permeation differences among racemic PL, S-PL enantiomer and R-PL enantiomer. CONCLUSION The developed RP-HPLC method is accurate, sensitive and efficient, and it is successfully applied to enantioselective transdermal permeation study of PL.