高效液相色谱法测定山银花提取物中7种有机酸的含量
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摘要
目的建立高效液相色谱法同时测定山银花提取物中7种有机酸的含量。方法准确称取一定量过筛的山银花提取物粉末经50%甲醇溶解并超声提取后,采用Acclaim 120 C18色谱柱(4.6 mm×250 mm,5μm),流动相为乙腈-0.4%磷酸溶液进行梯度洗脱,流速为1.0 m L/min,柱温为25℃,检测波长为327 nm,测定7种有机酸的含量。结果 7种有机酸在相应浓度区间线性良好(r>0.999),回收率为86.3%~97.7%,相对标准偏差<5%(n=6)。结论该方法操作简便、结果准确、灵敏度高,适用于山银花提取物中7种有机酸含量的测定。
Objective To establish a method for the determination of 7 kinds of organic acids in Lonicerae Flos extract by high performance liquid chromatography. Methods A certain amount of samples were weighed accurately, and dissolved with 50% methanol and extracted with ultrasounds. The samples were separated with a Acclaim 120 C18 column(4.6 mm×250 mm, 5 μm) at the flow rate of 1.0 m L/min by gradient elution using acetonitril and 0.4% phosphoric acid solution as the mobile phase. The temperature of the column oven was maintained at 25 ℃, the detection wavelength was 327 nm, and then the content of 7 kinds of organic acids was detected. Results The 7 kinds of organic acids showed a good linear correlation in certain concentration range(r>0.999), respectively. The average recoveries of 7 kinds of organic acids were 86.3%-97.7%, and the relative standard deviations were less than 5%(n=6). Conclusion This method is simple, accurate and highly sensitive, which is suitable for the determination of 7 kinds of organic acids in Lonicerae Flos extract.
Objective To establish a method for the determination of 7 kinds of organic acids in Lonicerae Flos extract by high performance liquid chromatography. Methods A certain amount of samples were weighed accurately, and dissolved with 50% methanol and extracted with ultrasounds. The samples were separated with a Acclaim 120 C18 column(4.6 mm×250 mm, 5 μm) at the flow rate of 1.0 m L/min by gradient elution using acetonitril and 0.4% phosphoric acid solution as the mobile phase. The temperature of the column oven was maintained at 25 ℃, the detection wavelength was 327 nm, and then the content of 7 kinds of organic acids was detected. Results The 7 kinds of organic acids showed a good linear correlation in certain concentration range(r>0.999), respectively. The average recoveries of 7 kinds of organic acids were 86.3%-97.7%, and the relative standard deviations were less than 5%(n=6). Conclusion This method is simple, accurate and highly sensitive, which is suitable for the determination of 7 kinds of organic acids in Lonicerae Flos extract.
引文
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[8]钟根秀,任琰,于志斌,等.我国植物提取物产业发展状况及建议[J].中国现代中药,2015,10:1087-1090.Zhong GX,Ren Y,Yu ZB,et al.Development situation of plant extract industry and policy suggestions[J].Mod Chin Med,2015,10:1087-1090.
[9]李华丽,袁君,郁荣华,等.HPLC法测定甜叶菊中6个酚酸类成分含量[J].药物分析杂志,2017,37(2):219-223.Li LH,Yuan J,Yu RH,et al.Determination of six phenolic acid in the leaves of Stevia rebaudiana bertoni by HPLC[J].Chin J Pharm Anal,2017,37(2):219-223.
[10]刘敏彦,高淑丽,刘丽华,等.HPLC法同时测定不同产地金银花和山银花中6种有机酸成分[J].中药材,2013,36(2):196-198.Liu YM,Gao SL,Liu LH,et al.Simultaneous determination of six organic acids in Lonicerae Flos and Lonicerae Flos in different habitats by HPLC[J].J Chin Mater,2013,36(2):196-198.
[11]李丹,李会军,高雯,等.一测多评法测定山银花提取物中6种绿原酸类成分[J].中国中药杂志,2014,39(6):2305-2310.Li D,Li HJ,Gao W,et al.Determination of six chlorogenic acids in Lonicerae flos extract by quantitative analysis multi-components with single marker[J].China J Chin Mater Med,2014,39(6):2305-2310.
[12]董俊兴.抗乙肝和艾滋病新药二咖啡酰奎尼酸进入临床[J].科技中国,2006,124(4):63.Dong JX.The new drug-diacinic acid anti-hepatitis b and AIDS entered the clinic[J].Sci Technol China,2006,124(4):63.
[13]吴卫华,康桢,欧阳东生.绿原酸的药理学研究进展[J].天然产物研究与开发,2006,18:691-694.Wu WH,Kang Z,Ouyang DS.Progresses in the pharmacology of chlorogenic acid[J].Nat Prod Res Dev,2006,18:691-694.
[14]张骏,杨小利,王红勇.绿原酸对心肌缺血-再灌注损伤的防护作用基础研究[J].中国药业,2015,17(24):3-5.Zhang J,Yang XL,Wang HY.Protective effect of chlorogenic acid against myocardial ischemia/reperfusion injury[J].China Pharm,2017,17(24):3-5.
[15]朱鹏,苗潇磊,陈勇.绿原酸、隐绿原酸和新绿原酸在中性和碱性p H条件下的降解动力学[J].药学学报,2016,51(1):122-126.Zhu P,Miao XL,Chen Y.Degradation kinetics of chlorogenic acid,cryptochlorogenic acid,and neochlorogenic acid at neutral and alkaline p H values[J].Acta Pharm Sin,2016,51(1):122-126.
[2]刘婧慧.金银花与山银花质量评价及安全性对比研究[D].北京:北京中医药大学,2014.Liu JH.Comparative study on quality evaluation and safety of Lonicera japonica and Lonicera macranthoides[D].Beijing:Beijing University of Chinese Medicine,2014.
[3]李泮霖,贺利利,李楚源,等.金银花和山银花抗急性口腔炎症作用比较[J].中山大学学报(自然科学版),2016,55(4):118-122.Li BL,He LL,Li CY,et al.Comparison of anti-inflammatory effects between Lonicerae japonicae flos and Lonicerae flos using an acute stomatitis model[J].Acta Sci Nat Univ Suny,2016,55(4):118-122.
[4]杨斌,丘岳,王柳萍,等.广西山银花绿原酸体外抗炎作用及分子机制研究[J].中国药理学通报,2009,25(4):542-545.Yang B,Qiu Y,Wang LP,et al.Studies on the anti-inflammatory molecular mechanism of chlorogenic acid extracted from Lonicera confusa DC in vitro[J].Chin Pharmacol Bull,2009,25(4):542-545.
[5]张英锋,张立艳,马子川,等.植物绿原酸的研究进展[J].化学教育,2011,32(4):1-6.Zhang YF,Zhang LY,Ma ZC,et al.Advancement of plant chlorogenic acid[J].J Chem Edu,2011,32(4):1-6.
[6]席利莎,木泰华,孙红男,等.绿原酸类物质的国内外研究进展[J].核农学报,2014,28(2):292-301.Xi LS,Mu TH,Sun HN,et al.Advances in research on chlorogenic acids at home and abroad[J].J Nucl Agric Sci,2014,28(2):292-301.
[7]王丽萍,郭栋,王果,等.中药绿原酸的研究进展[J].时珍国医国药,2011,4(22):961-963.Wang LP,Guo D,Wang G,et al.Advancement of chlorogenic acid in traditional Chinese medicine[J].Lishizhen Med Mater Med Res,2011,4(22):961-963.
[8]钟根秀,任琰,于志斌,等.我国植物提取物产业发展状况及建议[J].中国现代中药,2015,10:1087-1090.Zhong GX,Ren Y,Yu ZB,et al.Development situation of plant extract industry and policy suggestions[J].Mod Chin Med,2015,10:1087-1090.
[9]李华丽,袁君,郁荣华,等.HPLC法测定甜叶菊中6个酚酸类成分含量[J].药物分析杂志,2017,37(2):219-223.Li LH,Yuan J,Yu RH,et al.Determination of six phenolic acid in the leaves of Stevia rebaudiana bertoni by HPLC[J].Chin J Pharm Anal,2017,37(2):219-223.
[10]刘敏彦,高淑丽,刘丽华,等.HPLC法同时测定不同产地金银花和山银花中6种有机酸成分[J].中药材,2013,36(2):196-198.Liu YM,Gao SL,Liu LH,et al.Simultaneous determination of six organic acids in Lonicerae Flos and Lonicerae Flos in different habitats by HPLC[J].J Chin Mater,2013,36(2):196-198.
[11]李丹,李会军,高雯,等.一测多评法测定山银花提取物中6种绿原酸类成分[J].中国中药杂志,2014,39(6):2305-2310.Li D,Li HJ,Gao W,et al.Determination of six chlorogenic acids in Lonicerae flos extract by quantitative analysis multi-components with single marker[J].China J Chin Mater Med,2014,39(6):2305-2310.
[12]董俊兴.抗乙肝和艾滋病新药二咖啡酰奎尼酸进入临床[J].科技中国,2006,124(4):63.Dong JX.The new drug-diacinic acid anti-hepatitis b and AIDS entered the clinic[J].Sci Technol China,2006,124(4):63.
[13]吴卫华,康桢,欧阳东生.绿原酸的药理学研究进展[J].天然产物研究与开发,2006,18:691-694.Wu WH,Kang Z,Ouyang DS.Progresses in the pharmacology of chlorogenic acid[J].Nat Prod Res Dev,2006,18:691-694.
[14]张骏,杨小利,王红勇.绿原酸对心肌缺血-再灌注损伤的防护作用基础研究[J].中国药业,2015,17(24):3-5.Zhang J,Yang XL,Wang HY.Protective effect of chlorogenic acid against myocardial ischemia/reperfusion injury[J].China Pharm,2017,17(24):3-5.
[15]朱鹏,苗潇磊,陈勇.绿原酸、隐绿原酸和新绿原酸在中性和碱性p H条件下的降解动力学[J].药学学报,2016,51(1):122-126.Zhu P,Miao XL,Chen Y.Degradation kinetics of chlorogenic acid,cryptochlorogenic acid,and neochlorogenic acid at neutral and alkaline p H values[J].Acta Pharm Sin,2016,51(1):122-126.