HPLC法测定大鼠血浆中伏立康唑的浓度及其药代动力学研究

doi:10.5246/jcps.2025.10.068

摘要/Abstract

摘要：

采用高效液相色谱法检测大鼠血浆中伏立康唑的浓度, 并通过内标法进行更为精确的检测和药代动力学研究。以氟康唑为内标, 柱温30 ℃、流动相为甲醇-水梯度洗脱(0~2.5 min: 50%甲醇; 2.5~4 min: 50%~70%甲醇; 4 min后: 70%甲醇)、流速1.0 mL/min、检测波长254 nm、进样20 μL检测。大鼠按40 mg/kg灌胃伏立康唑后, 于不同时间采血, 得到伏立康唑血药浓度, 采用 DAS2.0软件对其进行拟合, 得到其药动学参数。血浆中伏立康唑浓度在0.2～40 mg/L内, 线性关系良好(R² = 0.9992), 提取回收率大于85%且小于115%, 其日内、日间RSD < 10％。伏立康唑大鼠体内的分布半衰期 t_1/2α和消除半衰期t_1/2β均为69.315 min, AUC_0–t为8040.73 min·mg/L。实验建立的HPLC法简便、快速、准确地测定了伏立康唑在大鼠体内的药物代谢过程, 该方法可用于临床批量检测伏立康唑血样。

关键词: 伏立康唑, HPLC, 药代动力学, 真菌感染

Abstract:

This study aimed to establish a reliable high-performance liquid chromatography (HPLC) method for determining Voriconazole concentrations in rat plasma, employing an internal standard approach to enhance accuracy. The pharmacokinetics of Voriconazole were also investigated. The method utilized Fluconazole as the internal standard, with gradient elution of a methanol-water mobile phase (0–2.5 min: 50% methanol; 2.5–4 min: 50%–70% methanol; after 4 min: 70% methanol). The analysis was performed at 30 °C with a flow rate of 1.0 mL/min, a detection wavelength of 254 nm, and a 20-μL injection volume. Following a single oral dose of Voriconazole (40 mg/kg), plasma concentrations were measured at various time points and analyzed using DAS2.0 software to calculate pharmacokinetic parameters. The method demonstrated excellent linearity (R² = 0.9992) over the concentration range of 0.2–40 mg/L. The extraction recoveries ranged from 85% to 115%, and intra-day and inter-day relative standard deviations (RSDs) were below 10%. Pharmacokinetic analysis revealed a distribution half-life of 69.315 min, an elimination half-life of 69.315 min, and an AUC_0–t of 8040.73 min·mg/L after oral administration at 40 mg/kg. The proposed HPLC method was simple, rapid, and precise, making it suitable for pharmacokinetic studies of Voriconazole in rats. Furthermore, this method offered potential applicability for clinical batch detection of Voriconazole in blood samples.

Key words: Voriconazole, HPLC, Pharmacokinetics, Fungal infections

Supporting:

刘晨曦, 刘欢, 陈天萍, 何诗怡, 胡茳玲, 蒋心惠. HPLC法测定大鼠血浆中伏立康唑的浓度及其药代动力学研究[J]. 中国药学(英文版), 2025, 34(10): 921-931.

Chenxi Liu, Huan Liu, Tianping Chen, Shiyi He, Jiangling Hu, Xinhui Jiang. Quantification of Voriconazole in rat plasma using HPLC and pharmacokinetic analysis[J]. Journal of Chinese Pharmaceutical Sciences, 2025, 34(10): 921-931.

图/表 9

Figure 1. Structure of Voriconazole.

Figure 2. Specificity test. (a) Blank plasma; (b) quality control samples; (c) mixed control solution; (d) plasma samples from Sprague-Dawley rats after intragastric administration.

Figure 3. Linear evaluation results.

Table 1. Extraction recovery results.

Table 2. Determination results of precision and accuracy of Voriconazole plasma samples (n = 5).

Table 3. Results of stability test of Voriconazole plasma samples (n = 5).

Table 4. Voriconazole concentration-time data in rat plasma.

Figure 4. Plasma concentration-time curve of rats after intragastric Voriconazole.

Table 5. Main pharmacokinetic parameters of rats after intragastric administration of Voriconazole.

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