飞蛾藤不同药用部位多种成分含量的比较

Comparison of the Contents of Multiple Constituents in Different Medicinal Parts of Porana racemosa

目的:建立同时测定飞蛾藤不同药用部位(茎、嫩枝、叶)中东莨菪苷、东莨菪内酯、绿原酸、隐绿原酸、新绿原酸、异绿原酸A、异绿原酸B、异绿原酸C等8种成分含量的方法,并进行比较。方法:采用高效液相色谱(HPLC)法。以Agilent TC-C18为色谱柱,以乙腈-0.1%磷酸溶液为流动相进行梯度洗脱,流速为1.0 mL/min,柱温为30℃,检测波长为345 nm,进样量为10µL。比较飞蛾藤茎、嫩枝、叶中上述成分的含量并采用Markerlynx XS软件进行主成分分析。结果:东莨菪苷、东莨菪内酯、绿原酸、隐绿原酸、新绿原酸、异绿原酸A、异绿原酸B、异绿原酸C检测质量浓度的线性范围分别为0.0764~7.64、0.0628~6.28、0.0908~9.08、0.0800~8.00、0.0576~5.76、0.0944~9.44、0.0860~8.60、0.0788~7.88 mg/L(r均大于0.999);精密度、稳定性(24 h)、重复性试验的RSD均小于2.0%;平均加样回收率分别为99.71%(RSD=1.36%,n=6)、100.39%(RSD=1.76%,n=6)、99.20%(RSD=1.75%,n=6)、100.04%(RSD=2.63%,n=6)、98.57%(RSD=1.99%,n=6)、99.68%(RSD=1.84%,n=6)、99.90%(RSD=1.88%,n=6)、99.76%(RSD=1.47%,n=6)。上述8种成分在茎中的平均含量分别为9.7253、1.2865、7.2713、1.3476、0.9977、0.7109、0.6563、0.3647 mg/g,在嫩枝中分别为0.6903、0.4117、4.3943、0.6396、0.5313、1.3927、0.9891、1.1292 mg/g,在叶中分别为1.1951、0.6911、27.9529、6.1734、1.4051、0.5497、0.2888、0.7942 mg/g。主成分分析结果显示,飞蛾藤不同部位可分为3类,其中飞蛾藤茎大部分聚集在得分图的第1象限,嫩枝聚集在第3象限,叶聚集在第4象限。结论:所建方法简便、重复性好,可用于飞蛾藤不同药用部位中东莨菪苷等8种成分的含量测定。飞蛾藤叶中新绿原酸、绿原酸和隐绿原酸的平均含量相对较高,嫩枝中异绿原酸B、异绿原酸A和异绿原酸C的平均含量相对较高,茎中东莨菪苷、东莨菪内酯的平均含量相对较高。

OBJECTIVE:To establish a method for the simultaneous determination of eight constituents,such as scopolin,scopoletin,chlorogenic acid,cryptochlorogenic acid,neochlorogenic acid,isochlorogenic acid A,isochlorogenic acid B,and isochlorogenic acid C,in different medicinal parts(stems,twigs and leaves)of Porana racemosa,and to compare the contents of eight constituents.METHODS:HPLC method was adopted.The determination was performed on Agilent TC-C18 column with mobile phase consisted of acetonitrile-0.1%phosphate acid(gradient elution)at the flow rate of 1.0 mL/min.The column temperature was 30℃,and the detection wavelength was set at 345 nm.The sample size was 10μL.The contents of above constituents in stems,twigs and leaves of P.racemosa were compared,and the principal component analysis was carried out by Markerlynx XS software.RESULTS:The linear range of scopolin,scopoletin,chlorogenic acid,cryptochlorogenic acid,neochlorogenic acid,isochlorogenic acid A,isochlorogenic acid B,and isochlorogenic acid C were 0.0764-7.64,0.0628-6.28,0.0908-9.08,0.0800-8.00,0.0576-5.76,0.0944-9.44,0.0860-8.60,0.0788-7.88 mg/L,respectively(all r>0.999).RSDs of precision,stability(24 h)and reproducibility tests were all lower than 2.0%.The average recoveries were 99.71%(RSD=1.36%,n=6),100.39%(RSD=1.76%,n=6),99.20%(RSD=1.75%,n=6),100.04%(RSD=2.63%,n=6),98.57%(RSD=1.99%,n=6),99.68%(RSD=1.84%,n=6),99.90%(RSD=1.88%,n=6),99.76%(RSD=1.47%,n=6),respectively.The average contents of above eight constituents were 9.7253,1.2865,7.2713,1.3476,0.9977,0.7109,0.6563,0.3647 mg/g in stems;those were 0.6903,0.4117,4.3943,0.6396,0.5313,1.3927,0.9891,1.1292 mg/g in twigs;those were 1.1951,0.6911,27.9529,6.1734,1.4051,0.5497,0.2888,0.7942 mg/g in leaves,respectively.The results of principal component analysis showed that different parts of P.racemosa could be divided into 3 categories.Among them,most of the stems of P.racemosa gathered in the first quadrant of score plot,all the twigs gathered in the third quadrant,and all the leaves gathered in the fourth quadrant.CONCLUSIONS:Established method is simple and reproducible,and can be used for the determination of 8 constituents in different medicinal parts of P.racemosa.The average contents of neochlorogenic acid,chlorogenic acid and cryptochlorogenic acid in the leaves of P.racemosa are relatively high;the contents of isochlorogenic acid B,isochlorogenic acid A and isochlorogenic acid C in the twigs are relatively high;the average contents of scopolin and scopoletin in the stems are also relatively high.