Simultaneous quantitation of seven alkaloids in processed Fuzi decoction by rapid resolution liquid chromatography coupled with tandem mass spectrometry

A rapid analytical method based on rapid resolution LC coupled with MS/MS was first established to quantify seven alkaloids in processed Fuzi decoction. The chromatographic method was optimized to allow simultaneous analysis of all analytes in 5 min and demonstrated good linearity (r > 0.9995), repeatability (RSD < 4.36%), intra‐ and interday precisions (RSD < 5.07%) with good accuracies (97.76–105.08%) and good recovery (95.0–107.5%) of seven alkaloids, namely higenamine, benzoylhypaconine, benzoylmesaconine, benzoylaconine, aconitine, hypaconitine, and mesaconitine. The LODs for these markers were in the range of 2.30–17.00 pg/mL. Quantitative analysis of the seven alkaloids in Baifupian decoction and Heishunpian decoction showed that the content of the seven marker chemicals varied significantly and concluded that the quality of Fuzi was greatly affected by different processed methods. The developed method could be used as a rapid, sensitive, and reliable approach for assessment of the quality of processed Fuzi and related decoction.     

Qualitative and quantitative analysis of the chemical constituents in Mahuang‐Fuzi‐Xixin decoction based on high performance liquid chromatography combined with time‐of‐flight mass spectrometry and triple quadrupole mass spectrometers

High‐performance liquid chromatography coupled with time‐of‐flight mass spectrometry (HPLC‐TOF/MS) and high‐performance liquid chromatography–triple quadrupole mass spectrometry (HPLC‐QQQ/MS/MS) were utilized to clarify the chemical constituents of Mahuang‐Fuzi‐Xixin Decoction. There are 52 compounds, including alkaloids, amino acids and organic acids were identified or tentatively characterized by their characteristic high resolution mass data by HPLC‐QQQ/MS/MS. In the subsequent quantitative analysis, 10 constituents, including methyl ephedrine, aconine, songrine, fuziline, neoline, talatisamine, chasmanine, benzoylmesaconine, benzoylaconine and benzoylhypaconine were simultaneously determined by HPLC‐QQQ/MS/MS with multiple reaction monitoring mode. Satisfactory linearity was achieved with wide linear range and fine determination coefficient (r > 0.9992). The relative standard deviations (RSD) of inter‐ and intra‐day precisions were <3%. This method was also validated by repeatability, stability and recovery with RSD <3% respectively. A highly sensitive and efficient method was established for chemical constituents studying, including identification and quantification of Mahuang‐Fuzi‐Xixin decoction.     

近红外光谱结合随机森林算法：一种快速有效的附子产地溯源策略

可靠的原产地认证方法对于保护指定产地的高价值中药材（例如道地药材、地理标志产品等）至关重要。附子作为著名的传统中药和川产道地药材，疗效显著，临床应用广泛，在国内外市场需求量很大。不同产地的附子疗效和价格有所不同，大众很难通过传统经验进行准确鉴别，基于植物代谢组学模式下的质谱检测技术，测试样本制备过程繁琐冗长、操作复杂、检测时间长，且重现性偏低。近红外光谱作为一种成熟、快速、无损的检测技术，被机器学习集成后为中药材在线质量监管和控制带来新途径。基于近红外光谱技术结合随机森林算法建立了一种不同产地附子无损鉴别模型。在四川、陕西和云南等主要栽培区域共采集了255份附子样本，采用傅里叶变换近红外光谱获得所有样本的漫反射光谱信息。采用单一和组合光谱预处理方式以消除光谱中的多种干扰，并筛选出最佳预处理方式，以此为输入指标建立随机森林模型。采用灵敏度、特异度和平衡精度等指标评价了模型的综合性能。结果表明：Savitzky-Golay平滑+多元散射校正为最佳预处理方式；仅采用全波长数据，RF模型对3组省级的样本的预测准确率超过了90%，预处理后预测准确率达98.39%；对于市/县一级样本，RF模型同样具有优秀的判别能力，准确率大于75%。模型对道地产区周边栽培区域的样本，识别率达100%。过滤出前100个特征波数，重新优化模型，模型对各市/县级区域的识别精度超过85%，尤其是对一些产自高原样本的识别能力得到了明显提升。研究中采用了环境友好型溯源策略，分析速度更快，样品损失更少，精度更高，为不同产地附子快速、高效的鉴别提供了新模式，为后续附子及其相关炮制品的鉴别和溯源提供了参考。     

UPLC‐ESI/MS determination of 17 active constituents in two categorized formulas of traditional Chinese medicine,Sanhuang Xiexin Tang and Fuzi Xiexin Tang: application in comparing the differences in decoctions and macerations

A rapid and sensitive UPLC‐ESI/MS method was established and validated to determine 17 active constituents (aconitine, hypaconitine, mesaconitine, benzoylaconine, benzoylhypaconine, benzoylmesaconine, berberine, palmatine, jatrorrhizine, coptisine, baicalein, wogonin, baicalin, wogonoside, emodin, aloe‐emodin and rhein) in Sanhuang Xiexin Tang (SXT) and Fuzi Xiexin Tang (FXT), which are two classic compound recipes from Xiexin Tang categorized formulas in traditional Chinese medicien. The separation was performed on a UPLC BEH C₁₈ column gradient eluted using acetonitrile and 0.1% formic acid as mobile phase. ESI/MS was operated in positive ([M + H]⁺) in selected ion recording mode for analysis of alkaloids and flavones, while in negative ([M ? H]^?) selected ion recording mode for anthraquinones. All of the 17 constituents exhibited good linearity in a relatively wide concentration ranges with the lowest limits of detection of 0.38 ng/mL. All of the relative standard deviation values of intra‐ and inter‐precisions and stabilities of 17 constituents were within 5%. The method was successfully applied to determine 17 active constituents in decoctions and macerations of SXT and FXT. The results indicated that different preparative methods resulted in significant diversity in concentrations of the 17 analytes. Herb–herb interaction appeared between aconitum alkaloids in Aconiti Lateralis Radix Preparata and another three herbs. Copyright © 2013 John Wiley & Sons, Ltd.     

Simultaneous determination of six constituents in Mahuang Fuzi Xixin by UPLC-PDA–MS/MS

Mahuang Fuzi Xixin (MFX), a classic recipe in traditional Chinese medicine, belongs to an exterior-relieving formula. For quality control of the MFX products, qualitative analysis using ultra-high performance liquid chromatography with diode-array detector–tandem mass spectrometry (UPLC-PDA–MS/MS) was undertaken. Six compounds from the MFX were simultaneously detected. Among them, astragalin and kaempferol 3-rutinoside were quantified through the UPLC–MS/MS method, while asarinin, sesamin, kakuol and methyleugenol were quantified through the UPLC-PDA method. This method can be applied to the quantitative determination of the six compounds in the MFX.     

大孔树脂对麻黄细辛附子汤生物碱成分静态吸附及解吸附性能研究

采用高效液相色谱法(HPLC)测定麻黄细辛附子(MXF)汤复方水提液中盐酸麻黄碱、盐酸伪麻黄碱、苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱含量,考察了4种大孔树脂(HPD–100,HPD–722,HPD–400,HPD–600型)对5种生物碱的静态吸附、解吸附性能,筛选出分离纯化麻黄细辛附子汤中生物碱的最佳树脂。结果显示,HPD–722型树脂对5种生物碱的吸附过程稳定且吸附量大,具有较高的解吸附率(87.26%),优于其它3种树脂。HPD–722型大孔树脂适用于富集麻黄细辛附子汤中的生物碱类化合物。