黄　攀，刘湘丹，马　铭，卢岳华

(1. 湖南师范大学化学生物学及中药分析教育部重点实验室, 中国 长沙　410081； 2.湖南中医药大学药学院，中国 长沙　410208)

HPLC法同时测定灌胃补肾通络方大鼠血浆中马钱苷、阿魏酸和二苯乙烯苷含量

黄攀1，刘湘丹2，马铭1，卢岳华1*

(1. 湖南师范大学化学生物学及中药分析教育部重点实验室, 中国 长沙410081； 2.湖南中医药大学药学院，中国 长沙410208)

摘要目的：建立灌胃补肾通络方大鼠血浆中马钱苷、阿魏酸和二苯乙烯苷的含量测定方法． 方法：以甲醇-乙酸乙酯(V甲醇∶V乙酸乙酯=3∶1)提取处理血浆，氮气吹干后流动相复溶进样．色谱条件: Johnson spherigel C18柱(250 mm×4.6 mm，5 μm)；流动相: 甲醇-0.01%乙酸水溶液，梯度洗脱；检测波长: 马钱苷为235 nm，阿魏酸和二苯乙烯苷为320 nm；流速: 1 mL/min；柱温: 30 ℃；进样量: 10 μL． 结果：空白血浆中内源性物质不干扰样品的测定．马钱苷、阿魏酸和二苯乙烯苷分别在0.03~3.00，0.03~3.00和0.02~2.00 mg/L范围内均有良好的线性关系(r≥0.999) ，检测限依次为0.020,0.015和0.010 mg/L；提取回收率分别大于79%、74%和58%，日间、日内精密度(RSD)均分别小于10%,7.3%和9.7%．结论：所建立的含量测定方法操作简便，灵敏，准确，适用于大鼠灌胃补肾通络方后血浆中马钱苷、阿魏酸和二苯乙烯苷的同时测定，为补肾通络方中有效成分药动学研究提供参考．

关键词补肾通络方；马钱苷；阿魏酸；二苯乙烯苷；血药浓度；高效液相色谱法

Simultaneous Determination of Loganin, Ferulic Acid and Stilbene Glucoside

in Rat Plasma after Ig Administered with Bushen Tongluo Formula by HPLC

HUANGPan1,LIUXiang-Dan2,MAMing1,LUYue-Hua1*

( 1. Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research, Ministry of Education of China,

Hunan Normal University, Changsha 410081, China;

2. Hunan University of Chinese Medicine, Changsha 410208, China)

AbstractObjectiveTo establish an HPLC method for extracting and determining the loganin, ferulic acid and stilbene glucoside in rat plasma after ig administered with Bushen Tongluo Formula. MethodsThe plasma was extracted and treated with a mixture of methanol and ethyl acetate (3∶1=V∶V), then plasma extract evaporated to dryness under a stream of nitrogen flow, and the residue was reconstituted in HPLC mobile phase to inject. The conditions of chromatography: the chromatographic separations was performed on Johnson spherigel C18column (250 mm×4.6 mm, 5 μm particle size) with a mobile phase composed of methanol-water (from 30∶70 to 50∶50,V/V) containing 0.01% glacial acetic acid at a flow of 1 mL/min. Loganin was determined at 235 nm, ferulic acid and stilbene glucoside were determined at 320 nm. The temperature of column was 30 ℃. The inject volume was 10 μL. ResultsThe method showed good linearity and no endogenous material interfered with the marked compounds. The linear range of loganin, ferulic acid and stilbene glucoside was 0.03~3.00,0.03~3.00 and 0.02~2.00 mg/L, respectively(r≥0.999), the limit of quantification was 0.020, 0.015, and 0.010 mg/L, the extraction recovery was over 79%, 74% and 58%, within-day and between-day precisions were less than 10%, 7.3% and 9.7%, respectively. ConclusionThe established method is accurate, sensitive and convenient for detecting blood concentration of loganin, ferulic acid and stilbene glucoside. It could provide a renference for studying pharmacokinetics of active ingredients in Bushen Tongluo Formula.

Key wordsBushen Tongluo Formula; loganin; ferulic acid; stilbene glucoside; blood concentration; HPLC

补肾通络方是临床常用有效治疗肾病综合征(Nephrotic syndrome, NS)的经验方，由制首乌、白术、当归、丹参、淮山、山茱萸、川芎、爵床、车前子、泽兰等药物组成，临床及实验研究表明其对NS肾虚夹瘀症有较……