已確效之高效液相層析法分離 Pindolol 對映異構體及其相關物質   全文下載

Validated Stability-Indicating HPLC Method for the Separation of Pindolol Enantiomers and Its Related Substances

本研究開發一簡單，快速且穩定之逆相液相層析法（RP-LC），於強迫降解之降解物中分離pindolol（PDL）及其相關物質。層析分離使用C18 Inertsil-3V管柱（250 mm × 4.6 mm內徑），移動相為20 mM磷酸二氫鈉，乙?混合液，pH 4.0，流速1.0 mL/min，檢測波長205 nm，管柱維持室溫。以正相液相層析法（nP-LC）分離pindolol對映異構體，使用chiral pack AD-H管柱（250 mm × 4.6 mm 內徑），流動相為正己烷：乙醇：二乙胺（860：140 mL：0.05%, v/v），流速0.9 mL/min，檢測波長215 nm。本方法經特異性、線性範圍、精密度、準確度、溶液安定性、穩定性、偵測極限（LOD）與定量極限（LOQ）之確效。逆相液相層析法之選擇性確效，以pindolol儲備溶液對酸、鹼、光氧化和熱降解等反應為主。PDL濃度範圍0.01 - 100 μg/mL（r2 = 0.9998）及0.001 - 50 μg/mL（r2 = 0.9982）時，分別於逆相液相層析法（RP-LC）及正相液相層析法（RP-LC）呈線性。降解物的波峰不會干擾純PDL。本方法之應用經分析PDL錠劑及檢測相關物質確認。分析結果有統計數據為證。"

A simple, rapid and stability-indicating reverse-phase liquid chromatographic (RP-LC) method was developed for the separation of pindolol (PDL) and its related substances in the presence of its degradation products generated from forced decomposition studies. The chromatographic separation was achieved on a C18 Inertsil ODS-3V column (250 mm × 4.6 mm i.d.), using a mobile phase of 20 mM sodium dihydrogen orthophosphate-acetonitrile containing orthophosphoric acid to maintain pH at 4.0. Gradient elution was used at a flow rate of 1.0 mL/min. The UV detector was operated at 205 nm and the column temperature was maintained at ambient. Separation of PDL enantiomers was achieved on normal phase liquid chromatography (NP-LC) using a chiral pack AD-H (250 mm × 4.6 mm i.d.) column, with mobile phase consisting of n-hexane : ethanol : diethylamine (860 : 140 mL : 0.05%, v/v). A flow rate of 0.9 mL/min and detection wavelength of 215 nm were used. The method was validated for specificity, linearity and range, precision, accuracy, solution stability, robustness, limit of detection (LOD) and limit of quantification (LOQ). Selectivity of the RP-LC method was validated by subjecting the stock solution of PDL to acidic, basic, photolysis oxidative and thermal degradation. The calibra-tion curve was found to be linear in the concentration range of 0.01 - 100 µg/mL (r2 = 0.9998) and 0.001 - 50 µg/mL (r2 = 0.9982) of PDL for the RP-LC and NP-LC method, respectively. The peaks of degradation products did not interfere with that of pure PDL. The utility of the developed method was examined for chemical and chiral purity by analyzing the tablets containing PDL and also by determining the related substances of PDL. The results of analysis were supported by statistical parameters.