超高效液相色谱-紫外检测法测定肾复康合剂中落新妇苷和芦丁的含量
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摘要
建立了超高效液相色谱(UPLC)测定肾复康合剂中主成分落新妇苷和芦丁的含量。采用反相高效液相色谱法对落新妇苷和芦丁进行色谱分离和快速定量测定。以ACQUITY UPLCTM C18色谱柱(2.1 mm×100 mm,1.7μm)为分离柱,柱温35℃;以乙腈-0.1%甲酸水溶液进行梯度洗脱;流速0.35 m L/min;进样量5μL;检测波长为204 nm。通过光谱图、保留时间和峰面积参数对落新妇苷和芦丁进行定性定量检测。落新妇苷在2～75μg/m L的范围内线性关系良好(R~2>0.998);方法检出限为1μg/m L,方法定量限为2μg/m L;芦丁在4～150μg/m L的范围内线性良好(R~2>0.996);方法检出限为2μg/m L,方法定量限为4μg/m L,完全满足检测需求。该方法色谱分离较好,分析速度较快,前处理简单,适用于肾复康合剂中落新妇苷和芦丁的定性定量检测。
This study was aimed to establish a high performance liquid chromatography(UPLC-UV)test method for the determination of astilbin and rutin in shenfukang mixture.UPLC was used to isolate and quantitatively determine of astilbin and rutin.ACQUITY UPLCTM C18 chromatographic column(2.1 mm×100 mm,1.7μm)was used as the separation column at column temperature 35℃;Gradient elution was performed with acetonitrile-0.1%formic acid solution;The flow rate is 0.35 m L/min,the injection volume is 5μL,and the detection wavelength is 204 nm.In this paper,the quantitative and qualitative analysis of astilbin and rutin was carried out by spectrogram and retention time and peak area parameters.In the range of 2～75μg/m L,the linear relationship of astilbin is good(R~2>0.998);the detection limit of the method is 1μg/m L,and the quantitative limit of the method is 2μg/m L;In the range of 4～150μg/m L,the linear relationship of rutin is good(R~2>0.996);The detection limit of the method is 2μg/m L(S/N>3),and the quantitative limit of the method is 4μg/m L(S/N>10),fully meet testing requirements.The method has good separation performance,fast analysis speed and simple pretreatment,suitable for the qualitative and quantitative detection of astilbin and rutin in shenfukang mixture.
This study was aimed to establish a high performance liquid chromatography(UPLC-UV)test method for the determination of astilbin and rutin in shenfukang mixture.UPLC was used to isolate and quantitatively determine of astilbin and rutin.ACQUITY UPLCTM C18 chromatographic column(2.1 mm×100 mm,1.7μm)was used as the separation column at column temperature 35℃;Gradient elution was performed with acetonitrile-0.1%formic acid solution;The flow rate is 0.35 m L/min,the injection volume is 5μL,and the detection wavelength is 204 nm.In this paper,the quantitative and qualitative analysis of astilbin and rutin was carried out by spectrogram and retention time and peak area parameters.In the range of 2～75μg/m L,the linear relationship of astilbin is good(R~2>0.998);the detection limit of the method is 1μg/m L,and the quantitative limit of the method is 2μg/m L;In the range of 4～150μg/m L,the linear relationship of rutin is good(R~2>0.996);The detection limit of the method is 2μg/m L(S/N>3),and the quantitative limit of the method is 4μg/m L(S/N>10),fully meet testing requirements.The method has good separation performance,fast analysis speed and simple pretreatment,suitable for the qualitative and quantitative detection of astilbin and rutin in shenfukang mixture.
引文
[1]国家药典委员会.中华人民共和国药典(2015年版一部)[S].National Pharmacopoeia Committee. Shenfukang Capsule Standard,Chinese Pharmacopoeia(1 part,2015)[S].
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[3]严爱娟,郭增喜,张文婷,等.土茯苓落新妇苷含量测定药典方法的改进[J].中药材,2016,8:1822-1823.Yan A J,Guo Z X,Zhang W T,et al. Improvement of test method for content of Rhizoma Smilacis glabrae and astilbin in Chinese Pharmacopoeia[J]. Chinese Traditional Medicine,2016,8:1822-1823.
[4]白梅,刘为萍,李素珍. HPLC测定不同产地土茯苓中落新妇苷和黄杞苷含量[J].药物分析杂志,2013,(8):1352-1354.Bai M,Liu W P,Li S Zh. Determination of content of astilbin and engelitin in Rhizoma Smilacis glabrae from different place[J]. Journal of Medicine Analysis,2013,(8):1352-1354.
[5]张笑颜,詹丽茵,曾祥腾.高效液相色谱法测定不同产地土茯苓药材中落新妇苷的含量[J].中国医药导报,2010,(7):90-91.Zhang X Y,Zhan L Y,Zeng X T. Using HPLC to test the content of astilbin in Rhizoma Smilacis glabrae from different place[J]. China Medicine Herald,2010,(7):90-91.
[6]隋欣,王人悦.落新妇苷稳定性的研究[J].食品研究与开发,2015,8:10-12.Sui X,Wang R Y. Study of astilbin stability[J].Food Study and Research,2015,8:10-12.
[2]农业部兽药评审中心.兽药研究技术指导原则[S].Center for Veterinary Drug Evaluation,Ministry of Agriculture.Guidance for Veterinary Drug Research Technology[S].
[3]严爱娟,郭增喜,张文婷,等.土茯苓落新妇苷含量测定药典方法的改进[J].中药材,2016,8:1822-1823.Yan A J,Guo Z X,Zhang W T,et al. Improvement of test method for content of Rhizoma Smilacis glabrae and astilbin in Chinese Pharmacopoeia[J]. Chinese Traditional Medicine,2016,8:1822-1823.
[4]白梅,刘为萍,李素珍. HPLC测定不同产地土茯苓中落新妇苷和黄杞苷含量[J].药物分析杂志,2013,(8):1352-1354.Bai M,Liu W P,Li S Zh. Determination of content of astilbin and engelitin in Rhizoma Smilacis glabrae from different place[J]. Journal of Medicine Analysis,2013,(8):1352-1354.
[5]张笑颜,詹丽茵,曾祥腾.高效液相色谱法测定不同产地土茯苓药材中落新妇苷的含量[J].中国医药导报,2010,(7):90-91.Zhang X Y,Zhan L Y,Zeng X T. Using HPLC to test the content of astilbin in Rhizoma Smilacis glabrae from different place[J]. China Medicine Herald,2010,(7):90-91.
[6]隋欣,王人悦.落新妇苷稳定性的研究[J].食品研究与开发,2015,8:10-12.Sui X,Wang R Y. Study of astilbin stability[J].Food Study and Research,2015,8:10-12.