Department of Biopharmaceutics, Shenyang Pharmaceutical University, No. 103, Wenhua Road, Shenyang 110016, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Feb 1;878(3-4):466-70. doi: 10.1016/j.jchromb.2009.11.041. Epub 2009 Dec 3.
A rapid, sensitive and selective ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method with hydrophilic interaction chromatography has been developed and validated for the simultaneous determination of didanosine and valdidanosine (L-valine amino acid ester prodrug of didanosine) in rat plasma. Solid-phase extraction (SPE) column was employed to extract the analytes from rat plasma, with high extraction recovery (>85%) for both didanosine and valdidanosine. The analytes were then separated by hydrophilic interaction chromatography (HILIC column) and detected by a triple-quadrupole mass spectrometry equipped with an electrospray ionization (ESI) source. The method was linear over the concentration ranges of 2-20,000 ng/mL for didanosine and 4-300 ng/mL for valdidanosine. The lower limit of quantitation (LLOQ) of didanosine and valdidanosine was 2 and 4 ng/mL, respectively. The intra-day and inter-day relative standard deviation (RSD) were less than 15% and the relative errors (RE) were all within 15%. Finally, the validated UPLC-MS/MS method was successfully applied to the pharmacokinetic study after either didanosine or valdidanosine orally administrated to the Sprague-Dawley rats.
建立并验证了一种快速、灵敏、专属性强的超高效液相色谱-串联质谱（UPLC-MS/MS）法，用于同时测定大鼠血浆中的双脱氧腺苷（ddI）和缬更昔洛韦（ddI 的 L-缬氨酸氨基酸酯前药）。采用固相萃取（SPE）柱从大鼠血浆中提取分析物，ddI 和缬更昔洛韦的提取回收率均>85%。采用亲水作用色谱（HILIC 柱）分离分析物，并用电喷雾电离（ESI）源的三重四极杆质谱进行检测。ddI 的浓度范围为 2-20,000ng/mL，缬更昔洛韦的浓度范围为 4-300ng/mL，方法均呈线性。ddI 和缬更昔洛韦的定量下限（LLOQ）分别为 2ng/mL 和 4ng/mL。日内和日间相对标准偏差（RSD）均<15%，相对误差（RE）均在 15%以内。最后，该验证后的 UPLC-MS/MS 方法成功应用于大鼠口服 ddI 或缬更昔洛韦后的药代动力学研究。
