大鼠口服黄体酮脂质纳米粒的血药浓度测定

目的采用高效液相色谱法测定大鼠血浆中黄体酮的浓度,以用于黄体酮脂质纳米粒的药动学研究.方法大鼠血样用乙酸乙酯提取,采用达那唑为内标,以Hypersil C18柱(150 mm×3.9 mm,5 μm) 为分析柱,甲醇∶水(60∶40)为流动相,流速0.6 ml/min,检测波长为240 nm.以去势大鼠为模型动物,应用建立的方法,测定大鼠口服黄体酮脂质纳米粒后的血浆药物浓度.结果黄体酮在0.02～2 μg/ml浓度范围内线性关系良好(r＝0.9999,n=3),定量限为(0.02±0.004)μg/ml(n=3),最低检测线为0.005 μg/ml(S/N≥3).高、中、低质控样品的日内、日间RSD均小于10%,平均提取回收率为90.5%,方法学回收率为93.4%～107.5%.血药浓度-时间曲线提示,脂质纳米粒延缓药物的达峰时间,并明显提高药物的生物利用度.结论本测定方法稳定,操作简便、快速、准确、灵敏,可用于黄体酮脂质纳米粒的药动学研究.

Determination of progesterone concentration in plasma after oral administration of progesterone-loaded lipid nanoparticles in rats

OBJECTIVE: To establish a RP-HPLC method for determination of plasma progesterone and to apply the method for pharmacokinetics study of progesterone-loaded lipid nanoparticles after oral administration in rats. METHODS: The plasma samples were collected from castrated rat after oral administration of progesterone-loaded lipid nanoparticles and extracted by acetic ether. The determination was performed on a Hypersil C18 column (150 mm X 3.9 mm , 5 microm) with a mobile phase consisting of methanol and water (60:40) at a flow-rate of 0.6 ml/min. The UV detector was at 240 nm and danazol was used as internal standard. RESULT: Good linearity was obtained over the range of 0.02-2 microg/ml progesterone in plasma(r=0.9999, n=3). The quantification limit was (0.02 +/-0.004) microg/ml(n=3) and the limit of detection was 0.005 microg.mL(-1)(S/N = or >3). The inter-and intra-day RSDs were all less than 10% for quality control samples at high-, medium- and low-concentrations. The average absolute recovery rate was 90.5 % and the average method recovery was in the range of 93.4 %-107.5%. The plasma concentration-time curves indicated that tmax was delayed after administration of progesterone-loaded lipid nanoparticles, and the bioavailability was increased significantly, compared with contrast solution. CONCLUSION: The method developed is stable, simple, rapid, accurate, sensitive and applicable for determining plasma concentrations of progesterone of progesterone-loaded lipid nanoparticles in pharmacokinetic studies.