基于双波长融合HPLC图谱的银屑病外用方中7种活性成分定量分析  

Quantitative analysis of seven active components in psoriasis external prescription based on two-wavelength fusion HPLC spectra

作　　者：洪亮 陈海明[2] 卢传坚[2] 李绍平[1] 赵静[1] HONG Liang;CHEN Hai-ming;LU Chuan-jian;LI Shao-ping;ZHAO Jing(State Key Laboratory of Quality Research in Chinese Medicine,University of Macao,Macao 999078,China;Guangdong Provincial Hospital of Chinese Medicine,Guangzhou 510120,China)

机构地区：[1]澳门大学中药质量研究国家重点实验室,中国澳门999078 [2]广东省中医院,广州510120

出　　处：《药物分析杂志》2022年第1期127-132,共6页Chinese Journal of Pharmaceutical Analysis

基　　金：国家自然科学基金(No.81673389);国家重点研发计划(2019YFC1711300);广东省重点领域研发计划(2020B1111110006);澳门科学技术发展基金项目(034/2017/A1和0017/2019/AKP);广州市科技局(201807010051);澳门大学科学研究基金项目(MYRG2018-00083-ICMS,MYRG2019-00128-ICMS和CPG2021-00009-ICMS)。

摘　　要：目的:测定银屑病外用方中7个主要活性成分(新落新妇苷、落新妇苷、新异落新妇苷、异落新妇苷、黄杞苷、异黄杞苷、柠檬苦素)含量。方法:采用Waters Xbrige BEH C;色谱柱(150 mm×4.6 mm,3.5μm),以水(A)-乙腈(B)为流动相,梯度洗脱(0~5 min,17%B;5~10 min,17%B→20%B;10~15 min,20%B;15~25 min,20%B→100%B),流速1.0 mL·min^(-1),柱温30℃,进样量5μL,检测波长为292 nm(0~19 min,新落新妇苷、落新妇苷、新异落新妇苷、异落新妇苷、黄杞苷、异黄杞苷)、210 nm(19~25 min,柠檬苦素),使用MATLAB软件编程,对CSV格式数据进行双波长融合。结果:新落新妇苷、落新妇苷、新异落新妇苷、异落新妇苷、黄杞苷、异黄杞苷和柠檬苦素线性范围分别为39.06~1250μg·mL^(-1)(r=0.9999)、26.25~840μg·mL^(-1)(r=0.9997)、18.64~596.4μg·mL^(-1)(r=0.9998)、6.31~202μg·mL^(-1)(r=0.9996)、2.53~80.8μg·mL^(-1)(r=0.9998)、1.33~42.6μg·mL^(-1)(r=0.9997)和5.08~162.4μg·mL^(-1)(r=0.9996),平均加样回收率(n=6)为96.1%~102.2%,RSD为0.48%~1.7%。银屑病外用方自制及医院提供的样品中新落新妇苷、落新妇苷、新异落新妇苷、异落新妇苷、黄杞苷、异黄杞苷和柠檬苦素的含量分别为1.06、1.21、0.87、0.30、0.09、0.07、0.22 mg·mL^(-1)和0.35、0.32、0.17、0.08、0.04、0.03、0.21 mg·mL^(-1)。结论:该方法能够快速、准确地测定银屑病外用方中7个活性成分含量,可用于银屑病外用方的质量控制。Objective:To determine the contents of 7 active components(neoastilbin,astilbin,neoisoastilbin,isoastilbin,engeletin,isoengeletin,limonin)in the psoriasis external prescription.Methods:The column wasWaters Xbrige BEH C;(150 mm×4.6 mm,3.5μm).The mobile phase consisted of solvent A(water)and solvent B(acetonitrile)with gradient elution(0-5 min,17%B;5-10 min,17%B→20%B;10-15 min,20%B;15-25 min,20%B→100%B).The flow rate was 1.0 mL·min^(-1).The column temperature was 30℃.Injection volume was 5μL.The detection wavelength for neoastilbin,astilbin,neoisoastilbin,isoastilbin,engeletin and isoengeletin were set at 292 nm during 0~19 min,and 210 nm for limonin during 19~25 min.MATLAB was used for two-wavelength fusion of CSV data.Results:The calibration curves of neoastilbin,astilbin,neoisoastilbin,isoastilbin,engeletin,isoengeletin and limonin were all in good linearity over the ranges of 39.06-1250μg·mL^(-1)(r=0.9999),26.25-840μg·mL^(-1)(r=0.9997),18.64-596.4μg·mL^(-1)(r=0.9998),6.31-202μg·mL^(-1)(r=0.9996),2.53-80.8μg·mL^(-1)(r=0.9998),1.33-42.6μg·mL^(-1)(r=0.9997)and 5.08-162.4μg·mL^(-1)(r=0.9996),respectively,and the average recoveries(n=6)were 96.1%~102.2%with RSD of 0.48%-1.7%.The contents of neoastilbin,astilbin,neoisoastilbin,isoastilbin,engeletin,isoengeletin and limonin in samples self-made and provided by a hospital were 1.06,1.21,0.87,0.30,0.09,0.07,0.22 mg·mL^(-1)and 0.35,0.32,0.17,0.08,0.04,0.03,0.21 mg·mL^(-1),respectively.Conclusion:This established method can determine the contents of 7 active ingredients in psoriasis external prescription quickly and accurately,and is also helpful for the quality control of psoriasis external prescription.

关 键 词：银屑病外用方 新落新妇苷 落新妇苷 新异落新妇苷 异落新妇苷 黄杞苷 异黄杞苷 柠檬苦素多波长融合 高效液相色谱法 

分 类 号：R917[医药卫生—药物分析学]