中文摘要：

目的建立同时测定给予丹红注射液主要组分后脑缺血再灌注损伤大鼠血浆中丹参素、原儿茶酸、香草酸、丹酚酸B和羟基红花黄色素A（HYSA）质量浓度的HPLC-DAD方法,为其药动学研究提供准确可靠的方法。方法制备大鼠大脑中动脉栓塞（MCAO）模型,iv丹红注射液主要有效成分丹参素、原儿茶醛、丹酚酸B和HYSA后,收集血样,采用Eclipse XDB-C18（150 mm×4.6 mm,5μm）色谱柱,以乙腈-0.4%磷酸水溶液为流动相进行梯度洗脱,检测波长280 nm用于检测丹参素、原儿茶酸、香草酸和丹酚酸B,403 nm用于检测HYSA,柱温为30℃,内标法定量。结果原儿茶醛在MCAO大鼠体内迅速被代谢,3 min即可检测到其代谢产物原儿茶酸和香草酸。丹参素、原儿茶酸、香草酸、丹酚酸B和HYSA的进样浓度分别在0.35-140 mg/L（R2=0.999 8）、0.15-60 mg/L（R2=0.999 0）、0.05-20 mg/L（R2=0.998 8）、0.25-100 mg/L（R2=0.999 6）、0.075-30 mg/L（R2=0.998 5）呈良好线性关系,平均回收率均在80%-120%,日内、日间RSD均小于10%,稳定性符合体内药物分析要求。大鼠iv给予丹红注射液主要有效成分后丹参素、原儿茶酸、香草酸、丹酚酸B和HYSA的主要药动学参数：AUC0-∞分别为（1143.862±230.840）、（427.024±59.293）、（135.785±47.631）、（418.631±66.242）、（288.788±87.809）mg·min/L,t1/2z分别为（71.496±29.067）、（82.379±26.279）、（40.331±6.006）、（125.164±59.709）、（177.577±112.836）min。结论所建立的HPLC-DAD分析方法专属性强,各成分分离度好,分析时长适宜,操作简单快速,可用于MCAO大鼠体内丹参素、原儿茶酸、香草酸、丹酚酸B、HYSA的同时测定及复方丹红注射液的药动学研究。

英文摘要：

Objective To develop an HPLC-DAD method for the simultaneous determination of danshensu, protocatechuic acid, vanillic acid, salvianolic acid B, and hydroxysafflor yellow A（HYSA） in cerebral ischemic injury of rats, in order to provide an accurate and reliable analytical method to study the pharmacokinetics. Methods The middle cerebral artery occlusion（MCAO） model was established, in which all the rats were iv injected with the active constituents in Danhong Injection simultaneously（danshensu, protocatechuic aldehyde, salvianolic acid B, and HYSA）. The analysis of the measured components＇ plasma concentration was carried at 30 ℃ on the reversed-phase column of Agilent Eclipse XDB-C18（150 mm × 4.6 mm, 5 μm） and eluted with acetonitrile and water containing 0.4% phosphate acid as mobile phase in gradient mode. Detection wavelengths were 280 nm for danshensu, protocatechuic acid, vanillic acid, salvianolic acid B and 403 nm for HYSA. The propyl p-hydroxybenzoate was used as the internal standard（IS）. Results Protocatechuic aldehyde rapidly metabolized in MCAO rats, but its metabolites, protocatechuic acid and vanillic acid, were easily detected. The linear ranges of danshensu, protocatechuic acid, vanillic acid, salvianolic acid B, and HYSA were 0.35—140（R2 = 0.999 8）, 0.15—60（R2 =0.999 0）, 0.05—20（R2 = 0.998 8）, 0.25—100（R2 = 0.999 6）, and 0.075—30 mg/L（R2 = 0.998 5）, respectively. The mean recoveries were between 80% and 120% and the RSD value of intra-day and inter-day was less than 10%. The results of stability meet the requirements for biopharmaceutical analysis. The main pharmacokinetic parameters of danshensu, protocatechuic acid, vanillic acid, salvianolic acid B and HYSA were as follows： AUC0~∞ were（1143.862 ± 230.840）,（427.024 ± 59.293）,（135.785 ± 47.631）,（418.631 ± 66.242）,（288.788 ± 87.809） mg·min/L and t1/2z were（71.496 ± 29.067）,（82.379 ± 26.279）,（40.331 ± 6.006）,（125.164 ± 59.709）,（177.577 ± 11