玄参药材中多类型成分的定量分析方法建立及其在脉络宁口服液中的应用

Establishment of quantitative method for analyzing multi-type components of Scrophulariae Radix and its application in Mailuoning Oral Liquid

目的建立超高效液相色谱-串联三重四极杆质谱(UPLC-QqQ-MS/MS)方法同时测定玄参Scrophulariae Radix中多类型成分的含量,并将此方法应用于脉络宁口服液(Mailuoning Oral Liquid,MOL),为玄参药材和MOL的质量评价提供依据。方法采用Waters HSS T3色谱柱(100 mm×2.1 mm,1.8μm),以0.1%甲酸水溶液和乙腈为流动相进行梯度洗脱。质谱采用电喷雾离子源(electrospray ionization,ESI),在多反应监测模式(multiple reaction monitoring,MRM)下对环烯醚萜苷、苯丙素苷、有机酸和核苷4类成分进行定量分析,结合化学计量学分析评价不同批次样品的质量。结果化学计量学结果表明,不同地区的玄参药材得到了有效区分,其质量差异可能与其生长环境相关。此外,4个年份(2019、2020、2021和2022年)的MOL各自聚成1类,可能是当年使用的药材质量相对稳定使得同一年份的MOL质量更相近;通过正交偏最小二乘-判别分析(orthogonal partial least squares-discriminate analysis,OPLS-DA)进一步筛选出不同年份MOL中异毛蕊花糖苷、对甲氧基肉桂酸、腺苷、1-(3-hydroxy-4-methoxyphenyl)ethyl-3-α-L-rhamnopyranosyl-6-caffeoyl-β-D-glucopyranoside、对羟基苯甲酸、阿魏酸和原儿茶酸7个差异性化学成分。结论该方法特异性强、准确度高且稳定性好,可为MOL中臣药玄参的药材选择及复方整体质量评价提供科学依据。

Objective One method of ultra-performance liquid chromatography-tandem triple quadrupole mass spectrometry(UPLC-QqQ-MS/MS)was developed for the simultaneous determination of multi-type components in Xuanshen(Scrophulariae Radix),and this method was subsequently applied to Mailuoning Oral Liquid(脉络宁口服液,MOL),providing a basis for their quality evaluation.Methods The chromatographic separation was performed on a Waters HSS T3 column(100 mm×2.1 mm,1.8μm)with the mobile phase of water containing 0.1%formic acid solution and acetonitrile under gradient elution.The electrospray ionization source(ESI)and multiple reaction monitoring(MRM)method were used for MS quantitative analysis of four types of components including iridoid glycosides,phenylpropyl glycosides,organic acids and nucleosides,and chemometrics was adopted to analyze the quantitative results of different batches of samples.Results The chemometric results showed that samples of Scrophulariae Radix of different regions could be distinguished effectively,and their quality difference might be related to the growing environment.In addition,MOL of four years(2019,2020,2021 and 2022)were grouped into one category each,respectively,suggesting that the quality of medicinal materials in the indicated year was relatively stable.Seven differential chemical components,including isoacteoside,p-methoxycinnamic acid,adenosine,1-(3-hydroxy-4-methoxyphenyl)ethyl-3-α-L-rhamnopyranosyl-6-caffeoyl-β-D-glucopyranoside,p-hydroxybenzoic acid,ferulic acid and protocatechuic acid,were figured out as the markers of quality control by orthogonal partial least squares-discriminate analysis(OPLS-DA)in MOL of different production years.Conclusion This new method is specific,accurate,and stable,which could provide a scientific basis for the selection of Scrophulariae Radix and the quality evaluation of MOL.