HPLC法同时测定复方穿心莲片中4种成分的含量
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摘要
目的:建立同时测定复方穿心莲片中穿心莲内酯、槲皮素-3-O-葡萄糖苷、脱水穿心莲内酯、山柰酚-3-O-葡萄糖苷含量的方法。方法:采用高效液相色谱法。色谱柱为Phenomenex C18,流动相为乙腈-0.1%磷酸(梯度洗脱),流速为1.0 ml/min,检测波长为225 nm(穿心莲内酯)、335 nm(槲皮素-3-O-葡萄糖苷)、254 nm(脱水穿心莲内酯和山柰酚-3-O-葡萄糖苷),柱温为25℃,进样量为10μl。结果:穿心莲内酯、槲皮素-3-O-葡萄糖苷、脱水穿心莲内酯和山柰酚-3-O-葡萄糖苷的检测进样量线性范围分别为0.111~2.770、0.064~1.605、0.086~2.138、0.050~1.260μg(r均为0.999 9);定量限分别为0.48、0.88、0.93、1.03 ng,检测限分别为0.79、1.15、0.96、1.07 ng;精密度、稳定性、重复性试验的RSD<2.0%;加样回收率分别为96.57%~100.30%(RSD=1.3%,n=9)、95.04%~98.87%(RSD=1.3%,n=9)、95.74%~102.92%(RSD=2.4%,n=9)、96.73%~100.95%(RSD=1.6%,n=9)。结论:该方法简单灵敏、准确可靠、重复性好,可用于同时测定复方穿心莲片中穿心莲内酯、槲皮素-3-O-葡萄糖苷、脱水穿心莲内酯、山柰酚-3-O-葡萄糖苷的含量。
OBJECTIVE:To establish a method for the simultaneous determination of andrographolide,quercetin-3-O-glucoside,dehydroandrographolide and kaempferol-3-O-glucoside in Compound chuanxinlian tablet. METHODS:HPLC was performed on the column of Phenomenex C18 with mobile phase of acetonitrile- 0.1% phosphoric acid aqueous(gradient elution)at a flow rate of 1.0 ml/min,the detection wavelength was 225 nm for andrographolide,335 nm for quercetin-3-O-glucoside and 254 nm for dehydroandrographolide and kaempferol-3-O-glucoside,column temperature was 25 ℃,and the volume injection was 10 μl. RESULTS:The linear range was 0.111-2.770 μg for andrographolide(r=0.999 9),0.064-1.605 μg for quercetin-3-O-glucoside(r=0.999 9),0.086-2.138 μg for dehydroandrographolide(r=0.999 9)and 0.050-1.260 μg for kaempferol-3-O-glucoside(r=0.999 9);the limits of quantitation were 0.48 ng,0.88 ng,0.93 ng and 1.03 ng,the limits of detection were 0.79 ng,1.15 ng,0.96 ng and1.07 ng,respectively;RSDs of precision,stability and reproducibility tests were lower than 2.0%;recoveries were 96.57%-100.30%(RSD=1.3%,n=9),95.04%-98.87%(RSD=1.3%,n=9),95.74%-102.92%(RSD=2.4%,n=9) and 96.73%-100.95%(RSD=1.6%,n=9),respectively. CONCLUSIONS:The method is sensitive,reliable and reproducible,and can be used for the simultaneous determination of andrographolide,quercetin-3-O-glucoside,dehydroandrographolide and kaempferol-3-O-glucoside in Compound chuanxinlian tablet.
OBJECTIVE:To establish a method for the simultaneous determination of andrographolide,quercetin-3-O-glucoside,dehydroandrographolide and kaempferol-3-O-glucoside in Compound chuanxinlian tablet. METHODS:HPLC was performed on the column of Phenomenex C18 with mobile phase of acetonitrile- 0.1% phosphoric acid aqueous(gradient elution)at a flow rate of 1.0 ml/min,the detection wavelength was 225 nm for andrographolide,335 nm for quercetin-3-O-glucoside and 254 nm for dehydroandrographolide and kaempferol-3-O-glucoside,column temperature was 25 ℃,and the volume injection was 10 μl. RESULTS:The linear range was 0.111-2.770 μg for andrographolide(r=0.999 9),0.064-1.605 μg for quercetin-3-O-glucoside(r=0.999 9),0.086-2.138 μg for dehydroandrographolide(r=0.999 9)and 0.050-1.260 μg for kaempferol-3-O-glucoside(r=0.999 9);the limits of quantitation were 0.48 ng,0.88 ng,0.93 ng and 1.03 ng,the limits of detection were 0.79 ng,1.15 ng,0.96 ng and1.07 ng,respectively;RSDs of precision,stability and reproducibility tests were lower than 2.0%;recoveries were 96.57%-100.30%(RSD=1.3%,n=9),95.04%-98.87%(RSD=1.3%,n=9),95.74%-102.92%(RSD=2.4%,n=9) and 96.73%-100.95%(RSD=1.6%,n=9),respectively. CONCLUSIONS:The method is sensitive,reliable and reproducible,and can be used for the simultaneous determination of andrographolide,quercetin-3-O-glucoside,dehydroandrographolide and kaempferol-3-O-glucoside in Compound chuanxinlian tablet.
引文
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[11]张慧晔,潘雪峰,徐小飞,等.复方穿心莲片的薄层鉴别研究[J].中成药,2015,37(2):460.
[12]胡强,周勇.HPLC法测定复方穿心莲片中脱水穿心莲内酯的含量[J].中国当代医药,2013,20(3):62.
[13]刘蕾,张瑜,韩光,等.HPLC法测定大鼠血浆中穿心莲内酯类衍生物的含量及其药动学研究[J].中国药房,2013,24(11):967.
[14]黄秀展,潘民儒,朱正英,等.HPLC法测定复方穿心莲片中穿心莲内酯和脱水穿心莲内酯的含量[J].中国社区医师,2012,14(7):28.
[2]国家药典委员会.中华人民共和国药典:一部[S].2015年版.北京:中国医药科技出版社,2015:268、350.
[3]朱艳玲.穿心莲化学成分和药理作用的研究进展[J].中国现代药物应用,2013,7(14):238.
[4]刘飞飞,范春林,黄晓群,等.穿心莲干浸膏HPLC特征指纹图谱研究[J].中药材,2007,38(7):1 505.
[5]李景华,许笑笑,赵炎葱,等.穿心莲二萜内酯有效部位化学成分的液质联用法鉴定及其初步药效学研究[J].中国中药杂志,2014,39(23):4 642.
[6]王莉宁,徐必学,林华容,等.蓝布正化学成分的研究[J].时珍国医国药,2009,20(4):798.
[7]杨威,郑明善,吕惠子.路边青化学成分的研究[J].延边大学医学学报,2013,36(1):32.
[8]欧阳玉祝,陈小东,唐红玉,等.路边青中总多酚的提取与分离研究[J].食品科学,2009,30(16):44.
[9]张飞.蓝布正的化学成分及质量控制研究[D].上海:上海交通大学,2012.
[10]李龙,翁恭峰,白玉春,等.复方穿心莲片质量标准的研究[J].广东药学院学报,2014,30(5):595.
[11]张慧晔,潘雪峰,徐小飞,等.复方穿心莲片的薄层鉴别研究[J].中成药,2015,37(2):460.
[12]胡强,周勇.HPLC法测定复方穿心莲片中脱水穿心莲内酯的含量[J].中国当代医药,2013,20(3):62.
[13]刘蕾,张瑜,韩光,等.HPLC法测定大鼠血浆中穿心莲内酯类衍生物的含量及其药动学研究[J].中国药房,2013,24(11):967.
[14]黄秀展,潘民儒,朱正英,等.HPLC法测定复方穿心莲片中穿心莲内酯和脱水穿心莲内酯的含量[J].中国社区医师,2012,14(7):28.