高效液相色谱-荧光检测法测定大鼠血浆中羟基酪醇及其药动学研究

Determination of Hydroxytyrosol in Rat Plasma by HPLC-FLD and its Pharmacokinetic Study

目的建立女贞子有效成分羟基酪醇在大鼠血浆中的高效液相色谱-荧光检测(HPLC-FLD)方法,研究其药动学特征。方法取SD大鼠6只,灌胃给予羟基酪醇(60 mg/kg),于给药前及给药后不同时间点眼眶采血,测定血浆中羟基酪醇的浓度,采用DAS 2.0软件对血药浓度-时间曲线进行统计距拟合,得出系列药动学参数。采用HPLC-FLD法,以水杨苷为内标,色谱柱:Kromasil C18(250 mm×4.6 mm,5μm),流动相:乙腈(A)-0.1%甲酸溶液(12:88),检测波长:激发波长(λEX)281 nm、发射波长(λEM)316 nm。结果血浆中羟基酪醇的线性范围为65.5~6550 (r=0.9990) ng/ml;主要药动学参数:t1/2α为(23.79±9.69) min,t1/2β为(38.33±15.65)min,AUC0-360min为(403.34±48.24)μg/ml·min, AUC0-∞为(407.89±49.62)μg/ml·min,Cmax为(6.00±1.01)μg/ml, Tmax为(13.33±2.58) min。结论该方法灵敏,简便,准确性高,选择性强,可用于羟基酪醇血药浓度的测定,羟基酪醇在大鼠体内的药动学符合二室模型特征。羟基酪醇在大鼠体内的吸收和消除均较快。

Objective To establish a sensitive and reliable HPLC-FLD method to determine the concentration of hydroxytyrosol of Fructus Ligustri Lucidi’s active ingredient in rat plasma and to investigate its pharmacokinetics. Methods Six rats were given intragastric gavage(i.g.) of 60 mg/kg) hydroxyltyrosol. Blood samples were collected via orbit vein at fixed time intervals before and after intragastric administration of hydroxytyrosol. The plasma concentrations were determined by HPLC-FLD. The datas were processed by the software of DAS2. 0. HPLC-FLD was performed on Kromasil C18(250 mm × 4.6 mm,5 μm) column with salicin as the internal standard. Mobile phase consisted of acetonitrile-0.1% formic acid solution(12:88).Results The linear ranges of hydroxytyrosol in rat plasma were 65.5-6550 ng/ml(r=0.9990).The excitation wavelength and the emission wavelength were set at 281 nm and 316 nm, respectively.The main pharmacokinetic parameters of t1/2α, t1/2β,AUC0-360 min, AUC0-∞,Cmax and Tmax were(23.79 ± 9.69) min,(38.33 ± 15.65)min,(403.34 ± 48.24) μg/ml · min,(407.89 ± 49.62) μg/ml · min,(6.00 ± 1.01) μg/ml,(13.33 ± 2.58) min. Conclusion The HPLC-FLD method is sensitive, simple, accurate, selective and suitable for plasma concentration determination. The pharmacokinetics of hydroxytyrosol were best fitted with two-compartment model after administration in rats, and hydroxytyrosol had a rapid absorption and elimination.