陈晨 王添敏 邸学 康廷国

【摘 要】 目的：对大鼠口服胡桃楸水提取物后血浆中没食子酸和丁香酸进行药代動力学研究，初步确定其药代动力学参数。方法：采用HPLC分析方法测定大鼠血浆中没食子酸和丁香酸的浓度。色谱柱：Agilent Poroshell 120 SB-C18（2.1×100 mm，2.7 μm），预柱：Agilent Poroshell 120 SB-C18（2.1×5 mm，2.7 μm）；流动相：0.2%甲酸水（v/v，A）- 0.2%甲酸乙腈（v/v，B），流速：0.3 mL/min，梯度洗脱条件为：0～3 min，0～5 % B；3～5 min，5～10 % B；5～13 min，10 % B，后运行时间为3 min，柱温为30 ℃。单次灌胃给予大鼠胡桃楸水提取物（以生药量计9.6 g/mL），灌胃剂量为1 mL/100 g，于给药前和给药后5、15、30、45、60、90、120、240、360和480 min分别取血0.2 mL，将血浆采用甲醇蛋白沉淀法处理后，取10 μL供HPLC分析。应用3P97软件处理数据，计算其药代动力学参数。结果：没食子酸在0.04～5 μg/mL范围内线性良好，最小检测限（LOQ）为0.006 μg/mL，最小定量限（LOD）为0.04 μg/mL；日内和日间精密度（RSD）分别为1.8 %～11.0 %和3.5 %～9.4 %，准确度（RE）分别为-0.7 %～1.3 %和-0.5 %～2.1 %；血浆提取回收率为81.2 %～94.4 %；药代动力学参数为Cmax=（0.64±0.05）μg/mL，Tmax=（61.80±8.03）min，T1/2=（184.21±12.73）min，AUC0-t=（96.37±10.33）μg·min/mL，AUC0-∞=（121.59±8.87）μg·min/mL。丁香酸在0.04～5 μg/mL范围内线性良好，最小检测限（LOQ）为0.008 μg/mL，最小定量限（LOD）为0.04 μg/mL；日内和日间精密度（RSD）分别为4.0 %～6.9 %和2.1 %～8.8 %，准确度（RE）分别为-1.2 %～4.9 %和-4.8 %～0.8 %；血浆提取回收率为84.9 %～93.6 %；药代动力学参数为Cmax=（0.43±0.04）μg/mL，Tmax=（30.67±0.92）min，T1/2=（99.63±5.96）min，AUC0-t=（40.33±2.42）μg·min/mL，AUC0-∞ =（47.02±3.29）μg·min/mL。结论：研究建立大鼠血浆中没食子酸和丁香酸浓度的高效液相色谱分析方法，经过方法学验证，该方法灵敏度高、重现性好、专属性强，符合生物样品分析的要求，并成功应用于大鼠血浆中没食子酸和丁香酸的分析。

【关键词】 胡桃楸；没食子酸；丁香酸；大鼠；药代动力学

【中图分类号】R285.5 【文献标志码】 A【文章编号】1007-8517（2018）04-0028-06

Abstract：Objective Made a pharmacokinetic research of gallic acid and syringic acid in rat plasma after orally administration of the water extraction of Juglans mandshurica Maxim. to preliminary confirm parameters of pharmacokinetics. Methods Used a HPLC analytical method to test concentrations of gallic acid and syringic acid in rat plasma. Column： Agilent Poroshell 120 SB-C18（2.1×100 mm， 2.7 μm）and Agilent Poroshell 120 SB-C18（2.1×5 mm， 2.7 μm）； mobile phases consist of ultra pure water （A） and acetonitrile （B） both with 0.2 % （v/v） formic acid at a flow rate of 0.3 mL/min， the following gradient elution was applied： from 0 % to 5 % B between 0 and 3 min， from 5 % to 10 % B between 3 and 5 min， 10 % B between 5 and 13 min， a 3 min post run time back to the initial mobile phase composition was used after each injection， column temperature was set at 30 ℃.Blood samples （0.2 mL） were collected before dosing and 5， 15， 30， 45， 60， 90， 120， 240， 360， 480 min after single dosing （crude herbal： 9.6 g/mL）， the dose of intragastric administration was 1 mL/100 g. After pre-treatment with protein precipitation method （methyl）， 10 μL was injected into HPLC system. The pharmacokinetic parameters were calculated by 3p97. Results Gallic acid had great linear at the range of 0.04～5 μg/mL， LOQ = 0.006 μg/mL， LOD = 0.04 μg/mL； the RSDs of precision were from 1.8 % to 11.0 % for inter-day assay and from 3.5 % to 9.4 % for intra-day. The accuracy were at the range of -0.7 %～1.3 % and -0.5 %～2.1 %， respectively； the extraction recoveries were from 81.2 % to 94.4 %； Cmax and Tmax were （0.64±0.05 μ）g/mL and （61.8±8.03）min， T1/2 were （184.21±12.73）min， AUC0-t and AUC0-∞ were （96.37±10.33）μg·min/mL and （121.59±8.87）μg@min/mL. Springic acid had great linear at the range of 0.04～5 μg/mL， LOQ = 0.008 μg/mL， LOD = 0.04 μg/mL； the RSDs of precision were from 4.0 to 6.9 % for inter-day assay and from 2.1 % to 8.8 % for intra-day. The accuracy were at the range of -1.2 %～4.9 % and -4.8 %～0.8 %， respectively； the extraction recoveries were from 84.9 % to 93.6 % ； Cmax and Tmax were （0.43±0.04）μg/mL and （30.67±0.92）min， T1/2 were （184.21±14.73）min， AUC0-t and AUC0-∞ were （40.33±2.42）μg·min/mL and （47.02±3.29）μg·min/mL. Conclusion The described HPLC method， with acceptable accuracy， which met all requirements in bioanalysis and was applied to analyze gallic acid and syringic acid in rat plasma successfully.

Keywords：Juglans mandshurica Maxim.； Gallic acid； Syringic acid； Rat； Pharmacokinetics

胡桃楸又名山核桃、核桃楸，为胡桃科胡桃属植物Juglans mandshurica Maxim.，分布于我国东北、华北以及朝鲜、日本等地区[1-2]。胡桃楸与同属植物胡桃（J. regic L.）功效类似，胡桃始载于宋代《开宝本草》，具有镇痛、清热解毒之功效。……