Analysis and evaluation of bioactive constituents from different parts of Epimedium brevicornum / 中国中药杂志

A comprehensive analytical method based on ultra-fast liquid chromatography coupled with triple quadrupole/linear ion trap tandem mass spectrometry(UFLC-QTRAP-MS/MS) was established for simultaneous determination of the content of 45 bioactive constituents including flavonoids, alkaloids, amino acids, phenolic acids, and nucleosides in Epimedium brevicornum. The multiple bioactive constituents in leaves, petioles, stems and rhizomes of E. brevicornum were analyzed. The gradient elution was performed at 30 ℃ in an XBridge~® C_(18) column(4.6 mm×100 mm, 3.5 μm) with 0.4% formic acid aqueous solution-acetonitrile as the mobile phase at a flow rate of 0.8 mL·min~(-1). Single factor experiment and response surface methodology were employed to optimize the extraction conditions. Multivariate statistical analyses including systematic cluster analysis(SCA), principal component analysis(PCA), partial least squares discriminant analysis(PLS-DA), and one-way analysis of variance(One-way ANOVA) were carried out to classify the samples from different parts and identify different constituents. Grey relation analysis(GRA) and entropy weight-TOPSIS analysis were performed to build a multi-index comprehensive evaluation model for different parts of E. brevicornum. The results showed that there was a good relationship between the mass concentrations of 45 constituents and the corresponding peak areas, with the correlation coefficients(r) not less than 0.999 0. The precision, repeatability, and stability of the established method were good for all the target constituents in this study, with the relative standard deviations(RSDs) less than 5.0%(0.62%-4.9%) and the average recovery of 94.51%-105.7%. The above results indicated that the bioactive constituents varied in different parts of E. brevicornum, and the overall quality followed the trend of leaves > petioles > rhizomes > stems. This study verified the rationality of the Chinese Pharmacopoeia(2020 edition) stipulating that the medicinal part of E. brevicornum is the leaf. Moreover, our study indicated that the rhizome had the potential for medicinal development. The established method was accurate and reliable, which can be used to comprehensive evaluate and control the quality of E. brevicornum. This study provides data reference for clarifying the medicinal parts and rationally utilizing the resources of E. brevicornum.

Simultaneous determination of multiple bioactive constituents in Abelmoschi Corolla by UFLC-QTRAP-MS/MS / 中国中药杂志

A comprehensive analytical method based on ultra-fast liquid chromatography coupled with triple quadrupole/linear ion trap tandem mass spectrometry(UFLC-QTRAP-MS/MS) was established for simultaneous determination of the content of 38 active components in Abelmoschi Corolla, including flavonoids, organic acids, nucleosides and amino acids, so as to investigate the effects of different harvesting and processing methods on multi-active components in Abelmoschi Corolla. The chromatographic separation was performed on a XBridg®C_(18) column(4.6 mm×100 mm, 3.5 μm) with(0.1% formic acid water) methanol-acetonitrile(1∶1) as the mobile phase for gradient elution at 30 ℃. The flow rate was 0.5 mL·min~(-1). The components were detected in a multiple-reaction monitoring(MRM) mode. The gray relational analysis(GRA) was used to comprehensively evaluate the multiple active components of Abelmoschi Corolla at different harvesting times and drying temperatures. The results showed that 38 components had a good linearity with correlation coefficients all above 0.999 0. The method featured a good precision, repeatability and stability with the relative stan-dard deviations(RSDs) of less than 5.0%. Recoveries ranged from 98.06% to 104.4% with RSD between 0.22% and 4.9%. The results of GRA indicated that a better quality in the samples collected on September 9 th. Samples dried at 90 ℃ had a better quality. The established method is accurate and reliable, and can be used to assess the internal quality of Abelmoschi Corolla. This study can provide basic materials for determining appropriate harvesting time and processing method of Abelmoschi Corolla.

Simultaneous Determination of Multiple Constituents in Panacis Japonici Rhizoma from Different Habitats by UFLC-QTRAP-MS/MS / 中国药学杂志

OBJECTIVE: To develop a method for the determination of saponins, amino acids and nucleosides in Panacis Japonici Rhizoma from different habitats by UFLC-QTRAP-MS/MS. METHODS: The chromatographic separation was performed on an XBridge C18 coulumn(4.6 mm×100 mm,3.5 μm) at 30 ℃ with gradient elution of 0.1% formic acid solution-0.1% formic acid acetonitrile, and the flow rate was 0.8 mL•min-1, using multiple-reaction monitoring(MRM) mode. RESULTS: The 33 constituents showed good linearity(r>0.999 0) in the range of the tested concentrations; the precision, repeatability and stability were good; the average recovery rates were between 96.95% and 101.8%, and the relative standard deviations were less than 5%. CONCLUSION: The established method is accurate and reliable, which can be used as a reference for the quality evaluation and control of Panacis Japonici Rhizoma.

Comparative analysis of multiple index constituents in Ophiopogonis Radix and Liriopes Radix / 中国中药杂志

An analytical method based on UFLC-QTRAP-MS/MS was established for simultaneous determination of thirty-three components including steroidal saponins, homoisoflavonoids, amino acids and nucleosides in Ophiopogonis Radix. Thirty-three target components of commercial medicinal materials of Maidong were comparative analysis. Synergi™ Hydro-RP 100 column (2.0 mm × 100 mm, 2.5 μm) was used with 0.1% formic acid solution-0.1% formic acid acetonitrile for gradient elution at a flow rate of 0.4 mL·min⁻¹. In addition, multiple reaction monitoring (MRM) mode was employed. The data were comprehensively processed and analyzed with hierarchical clustering analysis(HCA), principal component analysis(PCA) and partial least squares discriminant analysis(PLS-DA) methods. All components showed good linearity(>0.999 0) within the tested ranges. The average recoveries were between 96.23%-102.0%, and the relative standard deviation(RSD) were less than 5%. The results showed that there were significant differences in components between Ophiopogonis Radix and Liriopes Radix, with seven components obviously different. This method was useful for providing basis for the comprehensive evaluation and intrinsic quality control of Ophiopogonis Radix and Liriopes Radix , and may provide a new method reference for the identification of Ophiopogonis Radix and Liriopes Radix.

Stimultaneous determination of multiple bioactive constituents in Panacis Japonici Rhizoma processed by different methods and grey relational analysis / 中国中药杂志

A method, for determination of saponins, amino acids and nucleosides in Panacis Japonici Rhizoma of ultra fast liquid chromatography with triple quadrupole linear ion trap mass spectrometry (UFLC-QTRAP-MS/MS), was established to investigate the effect of different processing methods on the target components of Panacis Japonici Rhizoma. The chromatographic separation was performed on a XBridgeC₁₈(4.6 mm×100 mm, 3.5 μm) at 30 °C with a gradient elution of 0.1% formic acid solution-0.1% formic acid acetonitrile, and the flow rate was 0.8 mL·min⁻¹, using multiple-reaction monitoring (MRM) mode. The grey relational analysis was adopted for the analysis of different processing samples. The results showed that the thirty-three constituents were in a good linear range and the correlation coefficient was greater than 0.999 0; the precision, repeatability and stability were good; the average recovery rates were between 95.33% and 101.8%, and the relative standard deviations were less than 5%. The result of grey relational analysis showed that the complete rhizomes without peeling, which were adopted for the microwave dried method, had the best quality. The established method was accurate and reliable, which could be used to appraise the quality of Panacis Japonici Rhizoma. Our study may lay the way for the processing method of Panacis Japonici Rhizoma in optimization,normalization and standardization.

Comparative Analysis of Ginsenosides in Ginseng Radix et Rhizoma and Ginseng Radix et Rhizoma Rubra by UFLC-QTRAP-MS/MS / 中国药学杂志

OBJECTIVE To develop a comprehensive analytical method based on UFLC-QTRAP-MS/MS for simultaneous determination of protopanaxadiol [ginsenoside Rb1, Rc, Rb2, Rd, F2, 20(S)-Rg3, 20(R)-Rg3, CK], protopanaxatriol [ginsenoside Re, Rg1, Rf, 20(S)-Rg2, 20(S)-Rh1, 20(R)-Rg2, 20(R)-Rh1, F1] and oleanolic(ginsenoside Ro) in Ginseng Radix et Rhizoma and Ginseng Radix et Rhizoma Rubra. METHODS Under the optimized chromatographic conditions, good separation for seventeen target compounds was obtained on a SynergiTM Hydro-RP 100 column(2.1 mm×100 mm, 2.5 μm) at 40 ℃ with 0.1% aqueous formic acid (A)/acetonitrile (B) as the mobile phase by gradient elution at a flow rate of 0.4 mL·min-1. The target compounds were analyzed under multiple reaction monitoring (MRM) mode with an ESI source operated in negative ion mode, and principal component analysis (PCA) and hierarchical cluster analysis (HCA)were used for data processing. RESULTS The calibration curves of the 17 components had good linearity (r>0.999 0). The precision, repeatability and stability were all satisfying.The average recoveries of standard addition for the compounds were between 96.69% and 102.01%,and the relative standard deviations were less than 5%. The results of PCA and HCA showed that Ginseng Radix et Rhizoma and Ginseng Radix et Rhizoma Rubra were clearly distinguished.The main compositions with significant difference were ginsenoside 20(S)-Rg3, 20(R)-Rg3, 20(S)-Rh1, 20(R)-Rh1, and 20(R)-Rg2. CONCLUSION The established method could provide a new technique for the comprehensive evaluation and quality control of Ginseng Radix et Rhizoma and Ginseng Radix et Rhizoma Rubra, at the same time, it would pave the way for discovering the material basis contributing to the different properties and efficacies of the two medicinal materials.

Effects of Different Processing Methods on Lignans and Organic Acids in Schisandrae Chinensis Fructus / 中国药学杂志

OBJECTIVE: The influence of different processing methods on the quality of Schisandrae Chinensis Fructus was investigated based on simultaneous determination of multiple bioactive constituents combined with multivariate statistical analysis.METHODS: An analytical method based on UFLC-QTRAP-MS/MS was used for the simultaneous determination of fifteen components including lignans, such as schizantherin B, schisandrol B, schizandrin C, γ-schisandrin, deoxyschizandrin, schisantherin, schisandrin, schisanhenol, gomisin D, gomisin J, and angeloylgomisin H and organic acids, such as (S)-malic acid, D (-)-tartaric acid, protocatechuic acid, and quinic acid in Schisandrae Chinensis Fructus under different processing methods. Besides, clustering analysis and grey relation analysis(GRA) have been applied to comprehensively analyze and evaluate the results of different processing methods according to the content of 15 components.RESULTS: The results showed that fifteen components had good linearity, and correlation coefficients were more than 0.999 1. The method exhibited good precision, repeatability and stability. The average recoveries were between 96.64% and 99.96%, and the relative standard deviations were less than 5%. In addition, GRA results indicated that the quality of oven drying samples were better than the quality of oven drying after steaming samples. The quality of S7 was the best, followed by S10.CONCLUSION: The established method was accurate and reliable, which could be used to appraise the quality of Schisandrae Chinensis Fructus. Our study may lay the way for the processing method of Schisandrae Chinensis Fructus in optimization, normalization and standardization.

Simultaneous determination of eleven amino acids in Gynostemma pentaphyllum by UFLC-QTRAP-MS/MS / 中成药

AIM To establish a UFLC-QTRAP-MS/MS method for the simultaneous content determination of phenylalanine,isoleucine,methionine,valine,proline,tyrosine,aspartic acid,histidine,arginine,lysine and glutamic acid in Gynostemma pentaphyllum (Thunb.) Makino.METHODS The analysis of aqueous extract of G.pentaphyllum was performed on a Waters XBridge Amide column (2.1 mm × 100 mm,3.5 μm),with the mobile phase comprising of water-acetonitrile (containing 0.2％ formic acid) flowing at 0.6 mL/min in a gradient elution manner.RESULTS Eleven amino acids showed good linear relationships within their own ranges (r ＞0.998 9),whose average recoveries were 90.74％-103.05％ with the RSDs of 0.61％-5.00％.CONCLU-SION This accurate,stable and reproducible method can be used for the quality control of G.pentaphyllum.