Zhang Wei-Ku, Xu Jie-Kun, Hu Jie-Qing, Wang Su-Bo, Li Ping, Hiroshi Kurihara, Yao Xin-Sheng, Tang Bing-Hua
Department of Pharmacology, Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing 100029, China.
Zhongguo Zhong Yao Za Zhi. 2013 Mar;38(5):753-6.
To establish a method for the determination of theacrine in rat plasma after ig. administration of theacrine.
Blood sample was taken timely from the eyes canthus of rats. Plasma was isolated and the protein was precipitated by ethyl acetate. Then the plasma concentration of theacrine was determined with RP-HPLC. Caffeine was used as the internal standard. The chromatographic conditions were as follows: Phenomenex Luna C18 (4.6 mm x 250 mm, 5 microm) at 25 degrees C, a mixture of methanol-water (25: 75) as the mobile phase, at the flow rate of 1.0 mL x min(-1) and the detection wavelength of 290 nm.
The linear range of theacrine was 0.5-100 mg x L(-1) (R2 = 0.998 9). The lower limit of quantification was 0.5 mg x L(-1). The intra-day RSD was 1.49% 4.40% and inter-day RSD was 0.80% -10.27%. The average extraction recoveries of theacrine were 90.3% -95.8% at concentrations of 0.5, 5.0, 50 mg x L(-1). The main pharmacokinetic parameters after ig. administration of theacrine at concentration of 30 mg x kg(-1) were as follow: C(max) (35.45 +/- 30 2.68) mg x L(-1), t(max) (0.51 +/- 0.13) h, t1/2 (3.13 +/- 1.37) h, AUC(0-infinity) (2.65.39 +/- 94.71) mg x L(-1) x h.
The method has been confirmed to be simple, stable, reproducible and with high specificity, and can be used for the pharmacokinetic study of theacrine in rats.
建立灌胃给予茶氨酸后大鼠血浆中茶氨酸的测定方法。
从大鼠眼角及时采集血样。分离血浆，用乙酸乙酯沉淀蛋白质。然后采用反相高效液相色谱法测定血浆中茶氨酸的浓度。以咖啡因作为内标。色谱条件如下：Phenomenex Luna C18（4.6 mm×250 mm，5μm），柱温25℃，甲醇-水（25:75）混合液作为流动相，流速为1.0 mL·min⁻¹，检测波长为290 nm。
茶氨酸的线性范围为0.5 - 100 mg·L⁻¹（R² = 0.998 9）。定量下限为0.5 mg·L⁻¹。日内相对标准偏差为1.49% - 4.40%，日间相对标准偏差为0.80% - 10.27%。茶氨酸在0.5、5.0、50 mg·L⁻¹浓度下的平均提取回收率为90.3% - 95.8%。灌胃给予30 mg·kg⁻¹茶氨酸后的主要药代动力学参数如下：C(max)（35.45 ± 30 2.68）mg·L⁻¹，t(max)（0.51 ± 0.13）h，t1/2（3.13 ± 1.37）h，AUC(0 - infinity)（2.65.39 ± 94.71）mg·L⁻¹·h。
该方法经证实简单、稳定、可重复且特异性高，可用于大鼠体内茶氨酸的药代动力学研究。
