微乳液相色谱法同时测定大鼠灌服大花罗布麻叶提取物后血浆中4种黄酮类成分的含量
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篇名: 微乳液相色谱法同时测定大鼠灌服大花罗布麻叶提取物后血浆中4种黄酮类成分的含量
TITLE:
摘要: 目的:建立同时测定大鼠血浆中槲皮素-3-O-槐糖苷、罗布麻甲素、金丝桃苷、芸香苷4种黄酮类成分含量的方法。方法:取8只SD大鼠,ig大花罗布麻叶提取物混悬液,0.1 g/kg(以提取物计)。给药后1 h于眼眶采血1 mL,离心后采用微乳液相色谱法(MELC)测定4种黄酮类成分含量。色谱柱为Zorbax SB-C18,流动相为微乳体系[月桂醇聚氧乙烯醚-正丁醇-乙酸乙酯-三乙胺-水(体积比为2.5 ∶ 3.0 ∶ 1.7 ∶ 0.3 ∶ 92.5,pH=5.0)],流速为0.8 mL/min,柱温为30 ℃,检测波长为360 nm,进样量为10 μL,内标为岩白菜素。同时与使用常规有机溶剂为流动相的高效液相色谱法(HPLC,流动相为0. 2%磷酸水溶液-乙腈,梯度洗脱,其余条件同MELC法)进行相关比较。结果:大鼠血浆中槲皮素-3-O-槐糖苷、罗布麻甲素、金丝桃苷、芸香苷检测质量浓度线性范围为10~  1 000 ng/mL(r≥0.998 0), 定量下限(信噪比为10)均为10 ng/mL,精密度、稳定性、重复性试验的RSD均不超过7.5%(n=5),平均加样回收率在97.4%~98.5%之间(RSD均不大于5.3%,n=5),提取回收率均在88.7%~100.3%之间(RSD均不大于6.14%,n=5)。MELC与HPLC的方法学考察结果均符合药典规定,血药浓度测定结果相近,但与HPLC法比较,MELC法可缩短检测时间(10 min vs. 40 min),减少有机溶剂使用量(0.38 mL vs. 28 mL)。结论:本研究建立的MELC法快速简便、绿色环保,可用于大鼠血浆中大花罗布麻叶4种黄酮类成分的同时测定。
ABSTRACT: OBJECTIVE: To establish the method for the simultaneous determination of 4 flavonoids in rat plasma sample, such as quercetin-3-O-sophoroside, isoquercitrin, hyperin, rutin. METHODS: 8 SD rats were selected and given Poacynum hendersonii leaf extract suspension intragastrically 0.1 g/kg (calculated by extract). The blood sample 1 mL was collected from orbit 1 h after medication. The contents of 4 flavonoids were determined by microemulsion liquid chromatography (MELC) after centrifugation. The separation was performed on Zorbax SB-C18 column with mobile phase consisted of microemulsion [polyoxyethylene lauryl ether-n-butanol-ethyl acetate-triethylamine-water (volume ratio was 2.5 ∶ 3.0 ∶ 1.7 ∶ 0.3 ∶ 92.5, pH=5.0)] at the flow rate of 0.8  mL/min. The column temperature was set at 30 ℃, and detection wavelength was 360 nm. Sample size was 10 μL, and internal standard was bergenin. It was compared with HPLC method using organic solvent as mobile phase (mobile phase consisted of 0.2% phosphoric acid solution-acetonitrile, gradient elution, other condition same as MELC method). RESULTS: In rats plasma, the linear range of quercetin-3-O-sophoroside, isoquercitrin, hyperin, rutin were 10-1 000 ng/mL (r≥0.998 0). The limits of quantitation was 10 ng/mL (S/N=10). RSDs of precision,stability and reproducibility tests were all below 7.5% (n=5). Average sample recoveries were between 97.4%-98.5% and RSDs were less than 5.3% (n=5). Extraction recoveries were between 88.7%-100.3% (RSD≤6.14%, n=5). The methodological evaluation results of MELC and HPLC method were all in line with the regulations of pharmacopeia, and the results of blood concentration between them were similar. Compared with HPLC method, MELC method could shorten detection time (10 min vs. 40 min) and reduce the amount of organic solvent (0.38 mL vs. 28 mL). CONCLUSIONS: Established MELC method is rapid, simple and green, and can be used for 4 flavonoids from P. hendersonii leaf in rat plasma sample.
期刊: 2017年28卷第1期
作者: 庄鲁江,刘金岩,李欢欢,周军
AUTHORS: ZHUANG Lujiang,LIU Jinyan,LI Huanhuan,ZHOU Jun
关键字: 微乳液相色谱法;大鼠;血浆;大花罗布麻叶;黄酮类成分;含量测定
KEYWORDS: Microemulsion liquid chromatography; Rat; Plasma; Poacynum hendersonii leaf; Flavonoids; Content determination
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