離子對高效液相層析法同時檢測奈米微粒中 SN-38 內酯與羧酸酯   全文下載

Development and Validation of an Ion-pair HPLC Chromatography for Simultaneous Determination of Lactone and Carboxylate Forms of SN-38 in Nanoparticles

本研究開發簡易且具再現性之逆相離子對高效液相層析法，以等位沖提法配合紫外光偵測器，可同時檢測奈米微粒中 SN-38 內酯（L 型）與羧酸酯（C 型）。實驗採用 C18 管柱，以離子對溶液、磷酸二氫鉀緩衝液及乙睛 30:30:40 （v/v） 為移動相溶液，檢測條件為 265 nm，流速 1 mL/min。偵測結果，SN-38 C 及 L 型的尖鋒滯留時間分別為 2.9 ± 0.01 及 5.1 ± 0.01 分鐘。以線性迴歸數據分析，4 至 100 μg/mL 濃度範圍內，具良好的線性關係；L型及 C 型之迴歸係數為 0.9999 及 0.9998。本檢測方法具有高度準確性，專一性、線性、檢出及定量限量，精密度、準確度均符合國際醫藥法規協和會（International Conference on Harmonization, ICH）準則，可應用於 PLGANPs 中 SN-38 定量檢測。"

A simple and reproducible reversed-phase ion-pair high performance liquid chromatography (HPLC) method using isocratic elution with UV absorbance detection was developed and validated for the simultaneous determination of carboxylate (C) and lactone (L) forms of SN-38 (active metabolite of irinotecan) in nanoparticles (NPs). Reversed phase (RP) chromatography was performed on a C18 column with an ion-pair solution, KH2PO4 buffer solution and acetonitrile, under a mobile phase 30:30:40 (v/v) and at a flow rate of 1 mL/min. Detection was performed at 265 nm and sharp peaks were obtained for C and L forms of SN-38 at retention times of 2.9 ± 0.01 and 5.1 ± 0.01 min respectively. Linear regression analysis data for the calibration plot showed a good linear relationship between response and concentration in the range of 4 to 100 μg/mL; the regression coefficient was 0.9999 for L form and 0.9998 for C form. The optimized method showed good performance in terms of specificity, linearity, detection and quantitation limits, precision and accuracy in accordance with the International Conference on Harmonization (ICH) Q2 (R1) guidelines. This assay was demonstrated to be applicable for routine quantitation of SN-38 in PLGA NPs.