經確效之 HPLC 及 HPTLC 方法檢測 Ritonavir 原料藥粉及膠囊   全文下載

Validated Stability-Indicating HPLC and HPTLC Methods for the Determination of Ritonavir in Bulk Powder and in Capsules

本研究敘述兩種敏感且具再現性的方法，於降解產物中檢測抗反轉錄病毒藥物ritonavir含量。第一種是高效液相層析法（HPLC），自其降解物中分離ritonavir，層析管柱為逆相Agilent Eclipse XDB-C18 column（5-μm, 4.6 × 150 mm），移動相為乙?：0.05 M磷酸（55：45，v/v），流速1.0 mL/min。ritonavir滯留時間為4.82 ± 0.002 min。本方法利用二極管陣列檢測器作為波峰純度鑑定工具，濃度範圍1 - 500 μg/mL，波長210 nm時，以波峰面積和檢量線計算藥物含量。第二種方法係高效薄層層析法（HPTLC）， 240 nm斑點密度測量後，於Fluka TLC含螢光指示劑（254） nm矽膠鋁片上進行分離，移動相為乙?：水（1：2，V/V），以1 M的正磷酸溶液調節pH至5.0，ritonavir產生的對應斑點Rf值為0.41 ± 0.014；濃度範圍0.8 - 12.5 μg/spot時，線性迴歸方程式由校準數據最小平方產生。兩種方法的線性，精確性，準確度，偵測極限和定量極限，都經統計確效，並應用於Norvir膠囊檢測，沒有層析干擾現象。跟據ICH指引，兩種方法均可有效地自其降解物分離ritonavir，因此被認為是良好的穩定性指標方法。"

Two sensitive and reproducible methods were described for the quantitative determination of the antireteroviral drug ritonavir in the presence of its degradation products. The first method was based on high performance liquid chromatographic (HPLC) separation of the drug from its stress degradation products with the use of a reversed phase Agilent Eclipse XDB-C18 column (5 µm, 4.6 × 150 mm) and a mobile phase consisting of acetonitrile : 0.05 M phosphoric acid (55 : 45, v/v) at a flow rate of 1.0 mL/min. The retention time of the drug was found to be 4.82 ± 0.002 min. Quantification was achieved with diode array detection (DAD) at 210 nm based on peak area and a linear calibration curve in the concentration range of 1-500 µg/mL. The proposed method made use of diode array detection as a tool for peak purity and identification. The second method involved a high performance thin layer chromatographic (HPTLC) separation followed by densitometric measurement of the spots at 240 nm. The separation was carried out on Fluka TLC aluminium sheets of silica gel with fluorescent indicator (254) nm and the mobile phase was acetonitrile - water (1 : 2, v/v), adjusted to pH 5.0 using 1 M orthophosphoric acid solution . The proposed procedure gave compact spots for ritonavir (retention factor, Rf = 0.41 ± 0.014). The linear regression equation was generated by least-squares treatment of the calibration data in the range of 0.8-12.5 µg/spot. The reliability and analytical performance of the proposed methods, including linearity, range, precision, accuracy, detection and quantitation limits, were statistically validated. The proposed methods were applied to Norvir capsules and no chromatographic interference was observed. When ritonavir was subjected to stress conditions; according to ICH guidelines, the proposed methods could effectively separate the drug from its degradation products, and were thus considered as good stability-indicating procedures.