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OBJECTIVE To establish HPLC fingerprint of Shuangdong capsules, and to study the spectral effect relationship of its anti-inflammatory effect. METHODS The fingerprints of 15 batches of Shuangdong capsules were established by HPLC,and the similarity evaluation was carried out; the foot swelling model was established to investigate the anti-inflammatory activity of Shuangdong capsules. The gray correlation analysis method was used to construct the spectral effect relationship for the anti- inflammatory effect of Shuangdong capsules using the swelling rate of rat foot and the levels of malondialdehyde (MDA), superoxide dismutase (SOD), prostaglandin E2 (PGE2), interleukin-6 (IL-6), IL-8, IL-1β and tumor necrosis factor-α (TNF-α) in right hindfoot tissues as the pharmacodynamic indexes of anti-inflammatory effects. RESULTS Overall 15 batches of Shuangdong capsules identified 20 common peaks, the similarities were all greater than 0.97, and a total of 8 chromatographic peaks were identified. According to the gray correlation analysis, the correlation degrees between the peak area and the foot swelling rate and the levels of MDA, SOD, PGE2, IL-6, IL-8, IL-1β and TNF-α in 15 batches of Shuangdong capsules were 0.621 1- 0.783 5, 0.564 3-0.827 9, 0.581 0-0.845 3, 0.564 9-0.855 0, 0.583 1-0.856 4, 0.576 5-0.863 5, 0.564 1-0.838 0 and 0.572 5- 0.851 3, respectively. Among them, the chemical components represented by peak 4 (geniposidic acid), peak 10 (chlorogenic acid) and the chemical composition represented by peak 2 were strongly correlated with anti-inflammatory efficacy indicators. CONCLUSIONS In this study, HPLC fingerprints of 15 batches of Shuangdong capsules were successfully established. Among them, geniposidic acid, chlorogenic acid may be its anti-inflammatory ingredients.
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Objective:To establish the HPLC fingerprint of Bolbostemmatis Rhizoma standard decoction; To determine the three effective components with similar structure by quantitative analysis of multi-components by single marker (QAMS); To evaluate the quality of Bolbostemmatis Rhizoma standard decoction.Methods:HPLC was adopted to establish the fingerprints of 15 batches of Bolbostemmatis Rhizoma standard decoction. The Chromatographic column was Waters XBridge Phenyl (4.6 mm×250 mm, 5 μm). The mobile phase was acetonitrile-0.1% phosphoric acid solution with gradient elution. Cluster analysis (HCA) and principal component analysis (PCA) were conducted based on the relative peak area of common peaks. The same method as the fingerprint was used to establish QAMS of tubeimoside A, B, C on Bolbostemmatis Rhizoma standard decoction.Results:There were 14 common peaks in the fingerprint of Bolbostemmatis Rhizoma standard decoction. It was confirmed that the peak 3 was L-tryptophan, the peak 11 was tubeimoside B, the peak 12 was tubeimoside C, and the peak 13 was tubeimoside A. 15 batches of Bolbostemmatis Rhizoma standard decoction from different origins were divided into 3 categories by HCA and PCA. There was no significant difference between QAMS and the external standard method (ESM) through the system suitability inspection. Conclusion:This method is accurate, reliable and has good specificity, which can effectively evaluate the quality of Bolbostemmatis Rhizoma standard decoction.
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Objective:To compare the chemical composition of decoction and granules of Sangju Decoction; To provide a method for quality evaluation of Sangju Decoction.Methods:HPLC was used to establish fingerprints, and a comprehensive comparative study was conducted on the traditional decoction and formula granules of Sangju Decoction from four aspects: chemical composition type, fingerprint similarity, chemical pattern recognition analysis, and representative index component content.Results:The fingerprint similarity of the 10 batches of traditional decoction was >0.988. 35 peaks were identified and 12 peaks were identified as common peaks (neochlorogenic acid for peak 7, chlorogenic acid for peak 10, cryptochlorogenic acid for peak 11, 1,3-dicaffeoylquinic acid for peak 13, rutin for peak 17, lenoside A for peak 19, lignan for peak 20, isochlorogenic acid B for peak 24, ammonium glycyrrhizate for peak 25). The fingerprint similarity of the formulation pellets was >0.983, and 29 characteristic peaks were identified. Compared with the traditional decoction, some batches of the granules lacked peaks 14, 26, 27, 30, 32 and 34, and clustering analysis (CA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) could distinguish between the two. The contents of the 10 index components neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, forsythia ester glycoside A, grass glycosides, isochlorogenic acid B, 3,5-O-dicaffeoylquinic acid, forsythia glycosides, monkshood glycosides in the traditional soup were higher than that in the granules, and the contents of rutin and ammonium glycyrrhizate in the granules were higher than that in traditional decoction.Conclusions:The content and composition of traditional decoction and formula granules of Sangju Decoction are significantly different. The combination of fingerprinting and chemical pattern identification effectively can effectively evaluate the difference between traditional decoction and formula granules of Sangju Decoction, which can lay a foundation for the quality control and rational clinical application of formula granules of Sangju Decoction.
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Objective:To establish UPLC fingerprint method and 2 contents determination methods of Buddleja officinalis; To provide a reference for improving the quality control standard and evaluation of Buddleja officinalis from different habitats.Methods:UPLC method was used to establish the fingerprints of 17 batches of Buddleja officinalis. The similarity evaluation, clustering analysis, principal component analysis and orthogonal partial least squares discriminant analysis were used to compare the quality differences of Buddleja officinalis from different habitats. The contents of acteoside and linarin in Buddleja officinalis were determined.Results:There were 12 common peaks in UPLC fingerprints of Buddleja officinalis, six of which were identified as echinacoside, acteoside, cynaroside, isoacteoside, linarin, and apigenin. The fingerprint similarity of 17 batches of Buddleja officinalis was more than 0.9; Buddleja officinalis from different habitats were classified into 2 groups. Five differential markers were determined by OPLS-DA analysis. The order of significance was acteoside > peak 3 > echinacoside > isoacteoside > linarin. Edgeworthia chrysantha was identified by the method of fingerprint as counterfeit. The results of content determination showed that the content of Buddleja officinalis in Hubei and Sichuan was the high and stable.Conclusion:The method can effectively analyze the differences of Buddleja officinalis from different habitats, and provide reference for the quality control of Buddleja officinalis.
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ObjectiveTo improve the quality standard of Yuanhu Zhitong oral liquid in order to strengthen the quality control of this oral liquid. MethodThin layer chromatography(TLC) was used for the qualitative identification of Corydalis Rhizoma and Angelicae Dahuricae Radix in Yuanhu Zhitong oral liquid by taking tetrahydropalmatine, corydaline reference substances and Corydalis Rhizoma reference medicinal materials as reference, and cyclohexane-trichloromethane-methanol(5∶3∶0.5) as developing solvent, Corydalis Rhizoma was identified using GF254 glass thin layer plate under ultraviolet light(365 nm). And taking petroleum ether(60-90 ℃) -ether-formic acid(10∶10∶1) as developing solvent, Angelicae Dahuricae Radix was identified using a silica gel G TLC plate under ultraviolet light(305 nm). High performance liquid chromatography(HPLC) was performed on a Waters XSelect HSS T3 column(4.6 mm×250 mm, 5 μm) with acetonitrile(A)-0.1% glacial acetic acid solution(adjusted pH to 6.1 by triethylamine)(B) as the mobile phase for gradient elution(0-10 min, 20%-30%A; 10-25 min, 30%-40%A; 25-40 min, 40%-50%A; 40-60 min, 50%-60%A), the detection wavelength was set at 280 nm, then the fingerprint of Yuanhu Zhitong oral liquid was established, and the contents of tetrahydropalmatine and corydaline were determined. ResultIn the thin layer chromatograms, the corresponding spots of Yuanhu Zhitong oral liquid, the reference substances and reference medicinal materials were clear, with good separation and strong specificity. A total of 12 common peaks were identified in 10 batches of Yuanhu Zhitong oral liquid samples, and the peaks of berberine hydrochloride, dehydrocorydaline, glaucine, tetrahydropalmatine and corydaline. The similarities between the 10 batches of samples and the control fingerprint were all >0.90. The results of determination showed that the concentrations of corydaline and tetrahydropalmatine had good linearity with paek area in the range of 0.038 6-0.193 0, 0.034 0-0.170 0 g·L-1, respectively. The methodological investigation was qualified, and the contents of corydaline and tetrahydropalmatine in 10 batches of Yuanhu Zhitong oral liquid samples were 0.077 5-0.142 9、0.126 1-0.178 2 g·L-1, respectively. ConclusionThe established TLC, fingerprint and determination are simple, specific and reproducible, which can be used to improve the quality control standard of Yuanhu Zhitong oral liquid.
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OBJECTIVE To establish an HPLC fingerprint of Xiao’er resuqing oral liquid, and to determine the contents of twelve index components. METHODS HPLC method was adopted. The determination was performed on Venusil MP C18 column with mobile phase consisting of acetonitrile-0.1% phosphate aqueous solution (gradient elution) at a flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm, the column temperature was 30 ℃, the injection volume was 10 μL. HPLC fingerprint of Xiao’er resuqing oral liquid was established by using the Similarity Evaluation System of Chromatographic Fingerprint of TCM (2012 edition) to evaluate the similarity. The contents of 12 components were determined, including (R, S)-goitrin, 3,5-O-dicaffeoyl quinic acid, puerarin, forsythin, forsythoside A, chlorogenic acid, baicalin, saikosaponins d, wogonoside, baicalein, emodin and chrysophanol. RESULTS The similarity of HPLC fingerprints of 13 batches of Xiao’er resuqing oral liquid was greater than 0.97, and 14 common peaks were confirmed. The contents of the above 12 index components in 13 batches of Xiao’er resuqing oral liquid were as follows: 0.078-0.172, 1.564-2.736, 1.338-2.578, 0.426-0.872, 1.477-2.628, 1.396-2.447, 4.052-9.146, 0.367- 0.692, 1.974-4.674, 1.274-2.969, 0.085-0.167 and 0.155-0.307 mg/mL. CONCLUSIONS The established HPLC fingerprint and content determination methods have high accuracy and high specificity, which can be used for the quality evaluation of Xiao’er resuqing oral liquid.
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AIM To identify the chemical components of Longmu Qingxin Mixture by UPLC-Q-TOF-MS and study its material basis for the treatment of attention deficit hyperactivity disorder.METHODS The sample was detected by mass spectrometry in positive and negative ion mode on a Waters CORTECS? UPLC? T3 chromatographic column.The data were analyzed with Peakview 1.2 software and matched with the Natural Products HR-MS/MS Spectral Library 1.0 database,and the components were identified in combination with literature reports.The material basis of Longmu Qingxin Mixture for the treatment of attention deficit hyperactivity disorder was analysed according to the identified components.RESULTS Forty chemical components were identified,including 11 flavonoids,6 monoterpene glycosides,4 triterpene saponins,3 phenolic acids,6 alkaloids etc.,which mainly derived from Radix Astragali,Radix Paeoniae Alba,Radix Scutellariae,licorice root,Ramulus Uncariae cum,etc.,baicalein,formononetin,astragaloside Ⅳ and rhynchophylline may be the material basis for the therapeutic effect of Longmu Qingxin Mixture.CONCLUSION UPLC-Q-TOF-MS can quickly identify the chemical components of Longmu Qingxin Mixture.Flavonoids,triterpene saponins and alkaloids may be the material basis for Longmu Qingxin Mixture for the treatment of attention deficit hyperactivity disorder,which can provide the basis for its material basis research,quality standard establishment and pharmacological study of the dismantled formula.
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Objective To analyze the quality of 22 batches of Fritillariae thunbergii bulbus Formula Granules from 12 different manufacturers by using water-extraction reference substance of Fritillariae thunbergii bulbus(ZBM ERS ST)and water-extraction reference substance of Fritillariae hupehensis bulbus(HBBM ERS ST)as references.Methods Ethyl acetate-methanol-triethylamine-water(17∶1∶1∶0.5)was used as the developing solvent for high-performance thin-layer chromatography(HPTLC)fingerprint analysis.The high-performance liquid chromatography(HPLC)fingerprint analysis was performed on a Agilent Eclipse XDB-C18 column(4.6 mm×250 mm,5 μm)with the gradient mobile phase consisted of acetonitrile-0.03%diethylamine solution.The column temperature was set at 25℃and evaporative light-scattering detector was used.The determination was conducted according to standard test method for measurement of Fritillariae thunbergii bulbus Formula Granules(Guangdong PFKL00117).Results The results of HPTLC and HPLC analysis showed that there are significant differences among the 22 batches of Fritillariae thunbergii bulbus Formula Granules.There were 4 batches of Fritillariae thunbergii bulbus Formula Granules from 3 manufacturers among them showed fingerprint characteristics of Fritillariae hupehensis bulbus.The total amount of peimine and peiminine in the remaining 18 batches of Fritillariae thunbergii bulbus Formula Granules was 0.291-3.179 mg·g-1,which were quite different.Conclusion Currently,the quality of Fritillariae thunbergii bulbus Formula Granules on the market varies greatly.Standardized water-extract reference substance has better applicability for the analysis of the quality of Fritillariae thunbergii bulbus Formula Granules than the control medicinal materials.
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Objective To establish a method for simultaneous determination of HPLC fingerprint and multi-target ingredients in Atractylodis Macrocephalae Rhizoma(AMR),in order to provide reference for its quality control.Methods HPLC-DAD multi-wavelength switching method was used to establish fingerprint of AMR,similarity evaluation combined with hierarchical clustering analysis(HCA),principal components analysis(PCA)and discriminant analysis of partial least squares(PLS-DA)were used to carry out chemometric study.The contents of differential component such as atractylenolide Ⅰ,Ⅱ,Ⅲ and atractylon were determined simultaneously.Results The HPLC fingerprint of 37 batches of AMR was established.Nine common peaks were marked,and 4 of them were identified as atractylon,atractylenolide Ⅰ,Ⅱ,Ⅲ.The similarity degrees were between 0.539 and 0.996,the quality of AMR from different origin and different batches varies greatly.Atractylon,atractylenolide Ⅰ,Ⅱ,Ⅲ and one unknown component(peak 9)are the important factors affecting the quality of AMR.Conclusion The combination methods of HPLC fingerprint and simultaneous determinations of multiple components are simple,stable,accurate and reliable,which can provide reference for the quality evaluation of AMR and the improvement of quality standard,as well as lay a foundation for the basic research of its pharmacodynamic substances and related compound.
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OBJECTIVE To compare the changes of chemical components of Morus alba leaves, screen differential markers, and determine their contents, so as to provide reference for quality control of M. alba leaves before and after baked with honey. METHODS The fingerprints of M. alba leaves before and after baked with honey were established by high-performance liquid chromatography (HPLC). The common peaks of the fingerprints were identified and the similarity was evaluated. The differential markers of M. alba leaves before and after baked with honey were screened by principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) using common peak are of raw material and product baked with honey of M. alba leaves as index. The quantitative analysis was carried out. RESULTS Twenty-three and twenty-four common peaks were identified from the HPLC fingerprint spectra of ten batches of raw material and ten batches of product baked with honey of M. alba leaves, respectively. The similarities of HPLC fingerprints for raw material and product baked with honey of M. alba leaves were all greater than 0.97. The results of PCA showed that raw material and product baked with honey of M. alba leaves could be divided into two categories. The results of OPLS-DA showed that the variable importance in projection of peak 2, peak H (5- hydroxymethylfurfural), peak 1, peak 17 (isochlorogenic acid C) and peak 16 were all greater than 1. The average contents of differential marker of isochlorogenic acid C in raw material and product baked with honey of M. alba leaves were 0.093 6 and 0.127 8 mg/g, respectively; there was statistical significance (P<0.05). CONCLUSIONS Five differential markers such as isochlorogenic acid C are obtained. The content of isochlorogenic acid C in M. alba leaves is significantly increased after baked with honey.
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OBJECTIVE To analyze the compositional differences between Fructus Tritici Levis and Triticum aestivum, and to provide reference for identification and quality control of both. METHODS Twenty batches of Fructus Tritici Levis and three batches of T. aestivum were collected, and their fingerprints were acquired by high-performance liquid chromatography and the similarities were evaluated by the Evaluation System of Similarity of Chromatographic Fingerprints of Traditional Chinese Medicine (2012 version). Cluster analysis (CA), principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were performed to analyze the difference of Fructus Tritici Levis and T. aestivum from different regions, and the differential components were screened. The contents of the six identified components in Fructus Tritici Levis and T. aestivum were determined. RESULTS The similarities of the fingerprints of Fructus Tritici Levis ranged from 0.928 to 0.996, and the relative similarities of T. aestivum with Fructus Tritici Levis ranged from 0.761 to 0.773. A total of 19 common peaks were calibrated, and six components including linolenic acid, linoleic acid, 5-heptadecylresorcinol, 5-nonadodecylresorcinol, 5- heneicosylresorcinol, and 5-tricosylresorcinol were identified. The results of CA and PCA showed that Fructus Tritici Levis and T. aestivum could be clearly distinguished; the distribution of Fructus Tritici Levis from Anhui province was relatively concentrated. The results of OPLS-DA showed that linolenic acid, linoleic acid, and other six unknown compounds were the differential components between Fructus Tritici Levis and T. aestivum. The average contents of the six identified components in Fructus Tritici Levis were 0.100 9, 1.094 0, 0.005 1, 0.030 9, 0.098 2,and 0.024 8 mg/g, respectively; the contents of linolenic acid and linoleic acid in Fructus Tritici Levis were significantly higher than those in T. aestivum (P<0.05).CONCLUSIONS The established qualitative and quantitative methods are simple and reliable, and can be used for the identification and quality evaluation of Fructus Tritici Levis and T. aestivum. The identified differential components, such as linolenic acid and linoleic acid, can also provide clues for the differentiation and pharmacological study of Fructus Tritici Levis and T. aestivum.
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OBJECTIVE To analyze the compositional differences between Fructus Tritici Levis and Triticum aestivum, and to provide reference for identification and quality control of both. METHODS Twenty batches of Fructus Tritici Levis and three batches of T. aestivum were collected, and their fingerprints were acquired by high-performance liquid chromatography and the similarities were evaluated by the Evaluation System of Similarity of Chromatographic Fingerprints of Traditional Chinese Medicine (2012 version). Cluster analysis (CA), principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were performed to analyze the difference of Fructus Tritici Levis and T. aestivum from different regions, and the differential components were screened. The contents of the six identified components in Fructus Tritici Levis and T. aestivum were determined. RESULTS The similarities of the fingerprints of Fructus Tritici Levis ranged from 0.928 to 0.996, and the relative similarities of T. aestivum with Fructus Tritici Levis ranged from 0.761 to 0.773. A total of 19 common peaks were calibrated, and six components including linolenic acid, linoleic acid, 5-heptadecylresorcinol, 5-nonadodecylresorcinol, 5- heneicosylresorcinol, and 5-tricosylresorcinol were identified. The results of CA and PCA showed that Fructus Tritici Levis and T. aestivum could be clearly distinguished; the distribution of Fructus Tritici Levis from Anhui province was relatively concentrated. The results of OPLS-DA showed that linolenic acid, linoleic acid, and other six unknown compounds were the differential components between Fructus Tritici Levis and T. aestivum. The average contents of the six identified components in Fructus Tritici Levis were 0.100 9, 1.094 0, 0.005 1, 0.030 9, 0.098 2,and 0.024 8 mg/g, respectively; the contents of linolenic acid and linoleic acid in Fructus Tritici Levis were significantly higher than those in T. aestivum (P<0.05).CONCLUSIONS The established qualitative and quantitative methods are simple and reliable, and can be used for the identification and quality evaluation of Fructus Tritici Levis and T. aestivum. The identified differential components, such as linolenic acid and linoleic acid, can also provide clues for the differentiation and pharmacological study of Fructus Tritici Levis and T. aestivum.
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OBJECTIVE To establish high performance liquid chromatography (HPLC) fingerprint of Xiaohe syrup and determine the contents of 10 effective ingredients in them. METHODS With 12 batches of Xiaohe syrup as samples, ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was adopted with Athena C18 (250 mm×4.6 mm, 5 μm) as the chromatographic column, acetonitrile-0.1% phosphoric acid aqueous solution as mobile phase for gradient elution. The flow rate was 1.0 mL/min, and the detection wavelength was 210 nm. Similarity Evaluation System for Traditional Chinese Medicine Chromatographic Fingerprint (2012A version) was imported to establish the fingerprint of Xiaohe syrup and evaluate the similarity. The content determination was performed on ACQUITY UPLC BEH C18( 100 mm×2.1 mm, 1.7 μm) chromatographic column, with 0.01% formic acid acetonitrile-0.01% formic acid water as mobile phase for gradient elution at a flow rate of 0.4 mL/min; combined with high-resolution mass spectrometer, positive and negative ions were scanned with an electric spray ion source to determine the content of each main component in 12 batches of Xiaohe syrup. RESULTS A total of 33 common peaks were calibrated in 12 batches of samples, with similarities greater than 0.97; 10 chromatographic peaks were confirmed, namely flavonoid glycosides, paeoniflorin, ferulic acid, naringin, rosmarinic acid, neohesperidin, salvianolic acid B, tetrahydropalmatine, saikosaponin A, and saikosaponin D. The results of content determination showed that the above 10 components had good linear relationships within their respective mass concentration ranges (all R 2>0.999), with contents ranging from 0.35 to 0.64, 3.15 to 5.61, 0.11 to 0.17, 1.68 to 3.17, 1.59 to 1.90, 1.15 to 1.64, 0.78 to 1.48, 0.11 to 0.26, 0.06 to 0.13, and 0.33 to 0.61 mg/mL, respectively. CONCLUSIONS The main components of 12 batches of Xiaohe syrup are similar, but the contents vary; HPLC fingerprint and UPLC-MS/MS content determination method established in this study can be used for comprehensive quality evaluation of Xiaohe syrup.
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Objective To establish the HPLC fingerprint and pesticide residue detection methods for different kinds of rhubarb, and evaluate the quality of rhubarb comprehensively. Methods 20 batches of three types of rhubarb were collected and analyzed by high-performance liquid chromatography. The mobile phase was methanol-0.1 % phosphoric acid solution; gradient elution; column temperature of 35 ℃; detection wavelength of 254 nm; flow rate 1.0 ml/min. And cluster analysis was performed on the results. Direct extraction method was used and high-performance liquid chromatography-tandem mass spectrometry and gas chromatography-tandem mass spectrometry were established, 33 prohibited pesticides from different sources and origins of rhubarb were detected. Results The similarity among the fingerprint spectra of three sources of rhubarb, namely Rheum palmatum L., Rheum tanguticum Maxim.ex Balf., and Rheum officinale Bail1., and their control fingerprint spectra was>0.95. 20 batches of rhubarb samples were divided into 3 categories by cluster analysis. 33 prohibited pesticides were detected in rhubarb samples from different regions. Conclusion The quality of three kinds of rhubarb was significantly different. The established HPLC fingerprint and the method of banning agricultural residues were stable, reliable, simple and accurate, which could provide a basis for quality control evaluation of rhubarb.
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OBJECTIVE To identify the chemical constituents of Rhamni Songoricae Fructus and to establish their fingerprints and the method for simultaneous determination of four constituents to comprehensively evaluate the quality of Rhamni Songoricae Fructus. METHODS The chemical constituents in Rhamni Songoricae Fructus were qualitatively analyzed by ultra-performance liquid chromatography quadrupole-time-of-flight tandem mass spectrometry. The fingerprints of 15 batches of Rhamni Songoricae Fructus were established by HPLC and chemometric analysis was performed by using SPSS 26.0 and SIMCA 14.1 software; the contents of quercetin, kaempferol, kaempferide and emodin were determined by the same method. RESULTS A total of 35 constituents were identified, including 28 kinds of flavonoids, 5 kinds of anthraquinones and 2 kinds of organic acids. A total of 19 common peaks were identified in the HPLC fingerprints, recognizing quercetin, kaempferol, kaempferide and emodin. The similarities between HPLC fingerprints of 15 batches of samples and control chromatograms were greater than 0.9. The results of cluster analysis showed that 15 batches of samples were divided into 2 classes, of which S1-S5, S7 and S9 were one class and the rest were one class, similar to the results of principal component analysis. The results of the orthogonal partial least squares- discriminant analysis showed that the variable importance projections of peaks No. 2, 6, 1, 11 (quercetin), 3, 14, 8, 10, 19 (emodin), 5 were all greater than 1. The contents of quercetin, kaempferol, kaempferide and emodin ranged from 0.710 to 10.478 mg/g, 0.236 to 0.660 mg/g, 0.334 to 3.039 mg/g, and 0.261 to 0.504 mg/g. CONCLUSIONS The constructed chemical constituent identification, fingerprint and content determination methods are simple, feasible and reproducible, which combined with chemometric analysis can be used for comprehensive evaluation of the quality of Rhamni Songoricae Fructus.
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ObjectiveTo determine the pharmacodynamic substance basis of Epimedii Folium(EF) and Epimedii Wushanensis Folium(EWF) in promoting osteogenic differentiation, and to establish a method to analyze the material basis of Chinese materia medica based on the correlation between chemical fingerprint and cellular metabolomics. MethodThe chemical fingerprints of 15 batches of EF with 4 species and 3 batches of EWF were analyzed by ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive Orbitrap-MS), and partial least squares-discriminant analysis(PLS-DA) was used to analyze the peak areas of chemical fingerprints of samples. The effects of different samples on proliferative activity of MC3T3-E1 osteoblast precursors, as well as the activity of alkaline phosphatase(ALP) in osteoblasts were detected by cell counting kit-8(CCK-8) and enzyme-linked immunosorbent assay(ELISA). At the same time, UPLC-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) was used to analyze the effects of different samples on the metabolomics of MC3T3-E1 cells, then metabolic peak table of osteogenic differentiation cells was constructed, and pharmacodynamic index mean Y0 was introduced into the peak table. PLS was used to calculate mean Y0 of each group, and the mean Y0 was added to the peak table of chemical fingerprint to construct the correlation between chemical fingerprint and cell metabolome, the pharmacodynamic components of EF and EWF that promote bone differentiation were screened according to variable importance in the projection(VIP) value>1. The pharmacodynamic effects of EF and EWF were evaluated according to the mean Y0 of each group. ResultThe chemical fingerprints of EF with different origins and EWF were completely separated. Compared with the blank group, the activity of MC3T3-E1 cells in EF and EWF groups was significantly increased, the activity of ALP in the Epimedium brevicornu(Gansu province), E. koreanum and E. pubescens groups was significantly increased(P<0.05). The results of cell metabolomics showed that the blank group and the model group had an obvious trend of separation. EF with different origins and EWF had different distance from the model group, indicating that EF with different origins and EWF had different effect on promoting osteogenic differentiation. Chemical fingerprint-cell metabolomics integration analysis screened 9 components closely related to the efficacy of EF and EWF, including diphylloside B, epimedin C, icariin, baohuoside Ⅰ, yinyanghuo B, β-anhydroicaritin, magnoflorine, cryptochlorogenic acid and quercetin. E. koreanum had the strongest effect on promoting osteogenic differentiation. ConclusionThis study determined that the material basis of EF and EWF promoting osteogenic differentiation were mostly flavonoids, alkaloids and organic acids, which provided ideas and methods for the screening of pharmacodynamic components and the prediction of therapeutic effect of Chinese materia medica.
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ObjectiveTo establish a qualitative analysis method for the chemical constituents of the reference sample of Xiao Xumingtang, and to establish the fingerprint of 15 batches of Xiao Xumingtang, so as to evaluate the quality consistency among batches. MethodAccording to the key information of Xiao Xumingtang in the Key Information Table of Ancient Famous Classical Formulas(25 Formulas), the reference sample of this formula was prepared, and it was detected by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS). The chemical components were identified by self-constructed database, consulting relevant literature, and comparing with the reference substances, and the components were assigned by comparing with single drug samples and negative samples lacking single drug. The fingerprint of the reference sample of Xiao Xumingtang was established using high performance liquid chromatography(HPLC), and the common peaks were assigned and identified through single drug samples and negative samples lacking single drug. ResultBased on the information of MS fragments, relevant literature, and database retrieval, a total of 64 compounds were identified and inferred from the reference sample of Xiao Xumingtang, including 31 flavonoids, 8 terpenoids, 12 triterpenoid saponins, 2 phthalides, 3 phenylpropanoids, 2 gingerols, 5 alkaloids, and 1 cyanoside. Among them, 21 were derived from Scutellariae Radix, 10 from stir-fried Glycyrrhizae Radix et Rhizoma, 9 from Ginseng Radix et Rhizoma, 8 from Paeoniae Radix Alba, 4 from Saposhnikoviae Radix, 3 from Stephaniae Tetrandrae Radix, 3 from Chuanxiong Rhizoma, 2 from Aconiti Lateralis Radix Praeparata, 2 from Zingiberis Rhizoma Recens, 1 from Ephedrae Herba, and 1 from Armeniacae Semen Amarum. The established HPLC fingerprint of the reference sample of Xiao Xumingtang had 23 common peaks, among which, peaks 1 and 2 were derived from Paeoniae Radix Alba, peaks 3 and 7 from Saposhnikoviae Radix, peaks 4, 8 and 9 from Stephaniae Tetrandrae Radix, peaks 10, 17, 18, 20 and 21 from stir-fried Glycyrrhizae Radix et Rhizoma, peaks 11-16, 19 and 22 from Scutellariae Radix, peak 5 from Chuanxiong Rhizoma, peak 23 from Zingiberis Rhizoma Recens, peak 6 was the common component of stir-fried Glycyrrhizae Radix et Rhizoma and Scutellariae Radix. A total of 10 compounds including albiflorin(peak 1), paeoniflorin(peak 2), cimicifugoside(peak 3), 5-O-methylvisammioside(peak 7), baicalin(peak 11), sec-O-glucosylhamaudol(peak 13), oroxylin A-7-O-β-D-glucuronide(peak 15), wogonoside(peak 16), glycyrrhizic acid(peak 21) and 6-gingerol(peak 23) were identified. The similarities of 15 batches of reference samples were>0.999, indicating that the reference samples had good consistency. ConclusionThrough the identification of the chemical constituents in the reference sample of Xiao Xumingtang, it is clear that the composition of the samples is mainly composed of flavonoids and triterpenoid saponins. The established fingerprint can basically reflect the overall chemical characteristics of the reference sample of Xiao Xumingtang, which can provide a basis for the quality research of its compound preparations.
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ObjectiveTo analyze and determine the differential components of freeze-dried and sun-dried Panacis Quinquefolii Radix(PQR), and to compare the differences in their pro-angiogenic activities. MethodFingerprints of freeze-dried and sun-dried PQR were established based on ultra performance liquid chromatography(UPLC), and chemometrics methods such as principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were combined to determine the differential saponin composition of the two decoction pieces, and six representative saponins were selected and their contents in freeze-dried and sun-dried PQR were determined by UPLC. Transgenic zebrafish line Tg(fli1a∶EGFP) embryos fertilized for 24 h were selected, and different doses of 70% methanol extracts of freeze-dried and sun-dried PQR(10, 30 mg·L-1) were used to intervene in normal zebrafish and in a zebrafish model of intersegmental vascular(ISV) injury induced by vascular endothelial growth factor(VEGF) receptor tyrosine kinase inhibitor Ⅱ(PTK787), then the development of subintestinal vein(SIV) and ISV of zebrafish was observed, SIV diameter, mean number of crossings and mean number of germinations were determined, and the ISV vascular index was calculated, in order to compare the pro-angiogenic activities of the two decoction pieces. ResultThe similarity of the fingerprints of freeze-dried and sun-dried PQR decoction pieces was>0.950, and 17 common peaks were identified, of which 6 common peaks were designated as peak 6(ginsenoside Rg1), peak 7(ginsenoside Re), peak 8(ginsenoside Rb1), peak 11(ginsenoside Rc), peak 13(ginsenoside Rb2), and peak 16(ginsenoside Rd), respectively. A total of 11 differential saponin components were screened by PCA and OPLS-DA, indicating that there were some differences in the contents of the components in the two decoction pieces. The results of determination showed that the contents of ginsenosides Rg1, Re, Rb1 and Rb2 in freeze-dried PQR were higher than those in sun-dried PQR, while the contents of ginsenosides Rc and Rd were lower than those in sun-dried PQR(P<0.05, P<0.01). In the study of the pro-angiogenic effect on normal zebrafish embryos, compared with the blank group, and the SIV vessel diameter, mean germination rate and mean crossover rate were significantly higher in the high-dose groups of freeze-dried and sun-dried PQR(P<0.01), and the vessel diameter, mean numbers of crossings and germinations in the freeze-dried PQR group were higher than those of the sun-dried PQR group(P<0.05). In the study of the pro-angiogenic effect on zebrafish embryos with ISV injury, the development of ISV in the model group was significantly inhibited when compared with the blank group, compared with the model group, different dose groups of freeze-dried and sun-dried PQR could promote the growth and sprouting of ISV, and the number of normal blood vessels in the freeze-dried PQR group was significantly higher than that in the sun-dried PQR group at the same dosage(P<0.05). ConclusionFreeze-drying can effectively avoid the loss and secondary transformation of ginsenosides in PQR, and its angiogenic activity is better than that of sun-dried PQR, which can provide a reference for the production and development of high-quality PQR decoction pieces.
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ObjectiveTo investigate the material basis of homologous and heterogeneous effect of Aurantii Fructus Immaturus(AFI) and Aurantii Fructus(AF) based on the total statistical moment analysis and molecular connectivity index(MCI). MethodRelevant literature at home and abroad and Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) were consulted to establish the chemical composition database of AFI and AF, and set up their fingerprints by ultra-high performance liquid chromatography(UPLC), and the total statistical moments and similarity parameters of the fingerprint were calculated. According to MCI, all components of AFI and AF were divided into different component groups, the average values of 0-8th order(0χ-8χ) MCI of the common component groups of AFI and AF were calculated. ResultThe values of total zero-order moment(AUCT) of AFI and AF were (10.57±2.45)×106, (5.09±0.89)×106 μV·s, the values of total first-order moment(MCRTT) were (11.57±1.58), (12.10±1.29) min, the values of total second-order moments(VCRTT) were(24.49±2.30), (26.49±2.54) min2, respectively. It showed that qualitative and quantitative parameters of AFI and AF were significantly different. The components with high similarity such as neohesperidin, hesperidin and narirutin were screened as the common potential pharmacodynamic components of AFI and AF. The non-common components of AFI, such as alysifolinone and imperatorin, and the non-common components of AF, such as neoeriocitrin and isosakuranin, with high similarity were screened out as potential heterogeneous components of AFI and AF. The composition groups of AFI and AF were classified into six categories, and the similarities between the composition groups of AFI and AF and the total constituents were 0.872-0.979 and 0.918-0.997, the average values of 0χ-8χ MCI of alkaloids in AFI and AF were 3.65 and 3.14, the average values of 0χ-8χ MCI of flavonoids were 8.47 and 8.47, the average values of 0χ-8χ MCI of volatile oils were 2.71 and 3.48, respectively. It showed that there were some differences in MCI of chemical constituents(groups) between AFI and AF. ConclusionThe chemical constituents(groups) of AFI and AF not only differ in content and species, but also in structural characteristics and structure-activity relationship, which can provide a basis for further explaining the scientific connotation of homologous and heterogeneous effect of AFI and AF.
ABSTRACT
Objective To establish the method of fingerprint and content determination of multi-component for Xiangsha Yangwei pill by gas chromatography(GC).Methods The GC fingerprint of Xiangsha Yangwei pill was found,and the peak attribution was carried out.The contents of limonene,eucalyptol,camphor,borneol,bornyl acetate,patchouli alcohol,pogostone,and α-cyperone were determined.Results The fingerprint similarity of 56 batches of Xiangsha Yangwei pill were 0.33-0.99,28 common peaks were confirmed,and 14 known components were identified.Limonene,eucalyptol,camphor,borneol,bornyl acetate,patchouli alcohol,pogostone and α-cyperone showed good linearity within the determined ranges(14.30-286.08,24.52-490.44,16.14-322.88,9.40-187.95,15.39-307.83,25.78-515.60,19.95-398.90,and 24.87-497.30 μg·mL-1).The average recoveries were 101.20%,97.90%,93.97%,94.23%,102.94%,100.54%,99.16%,and 98.31%;with the RSDs were 2.41%,1.48%,1.65%,2.00%,1.93%,2.30%,2.07%,and 2.38%,respectively.The concentrations of eight components were 0.2-959.1,0.3-420.4,1.0-542.6,0.0-64.5,0.0-364.2,0.0-339.6,0.0-130.7,0.0-82.0 μg·g-1,respectively.Conclusion The fingerprint and multi-component determination method can be used for the quality control and evaluation of Xiangsha Yangwei pill.