Simultaneous determination of eleven volatile components in Cinnamomi Oleum by GC-MS / 中国中药杂志

A gas chromatography-triple quadrupole mass spectrometry(GC-MS) method was established for the simultaneous determination of eleven volatile components in Cinnamomi Oleum and the chemical pattern recognition was utilized to evaluate the quality of essential oil obtained from Cinnamomi Fructus medicinal materials in various habitats. The Cinnamomi Fructus medicinal materials were treated by water distillation, analyzed using GC-MS, and detected by selective ion monitoring(SIM), and the internal standards were used for quantification. The content results of Cinnamomi Oleum from various batches were analyzed by hierarchical clustering analysis(HCA), principal component analysis(PCA), and orthogonal partial least squares-discriminant analysis(OPLS-DA) for the statistic analysis. Eleven components showed good linear relationships within their respective concentration ranges(R~2>0.999 7), with average recoveries of 92.41%-102.1% and RSD of 1.2%-3.2%(n=6). The samples were classified into three categories by HCA and PCA, and 2-nonanone was screened as a marker of variability between batches in combination with OPLS-DA. This method is specific, sensitive, simple, and accurate, and the screened components can be utilized as a basis for the quality control of Cinnamomi Oleum.

Dynamic changes of total acid and bacterial communities during the traditional fermentation of Hong Qu glutinous rice wine

Background: Hong Qu glutinous rice wine (HQGRW) is brewed under non-aseptic fermentation conditions, so it usually has a relatively high total acid content. The aim of this study was to investigate the dynamics of the bacterial communities and total acid during the fermentation of HQGRW and elucidate the correlation between total acid and bacterial communities. Results: The results showed that the period of rapid acid increase during fermentation occurred at the early stage of fermentation. There was a negative response between total acid increase and the rate of increase in alcohol during the early fermentation stage. Bacterial community analysis using high-throughput sequencing technology was found that the dominant bacterial communities changed during the traditional fermentation of HQGRW. Both principal component analysis (PCA) and hierarchical clustering analysis revealed that there was a great difference between the bacterial communities of Hong Qu starter and those identified during the fermentation process. Furthermore, the key bacteria likely to be associated with total acid were identified by Spearman's correlation analysis. Lactobacillus, unclassified Lactobacillaceae, and Pediococcus were found, which can make significant contributions to the total acid development (| r| N 0.6 with FDR adjusted P b 0.05), establishing that these bacteria can associate closely with the total acid of rice wine. Conclusions: This was the first study to investigate the correlation between bacterial communities and total acid during the fermentation of HQGRW. These findings may be helpful in the development of a set of fermentation techniques for controlling total acid.

Hplc fingerprint and chemical pattern recognition of valeriana jatamansi / 中国药学杂志

OBJECTIVE: To establish the HPLC fingerprint of Valeriana jatamansi and provide a reference for its effective quality control. METHODS: The HPLC-DAD analysis was performed on Diamonsil C18 column (4.6 mm×250 mm, 5 μm), with acetonitrile (A)-0.1% formic acid (B) solution as the mobile phase for gradient elution, the detection wavelength was set at 327 nm (0-33 min) and 256 nm (33-90 min), the flow rate was 1.0 mL•min-1, and the column temperature was maintained at 30 ℃. The fingerprints of 25 batches of Valeriana jatamansi samples were analyzed by similarity analysis, hierarchical clustering analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (PLS-DA). RESULTS: The fingerprints of 25 batches of Valeriana jatamansi samples were established. There were 36 common peaks in the fingerprints and nine common peaks were identified by reference substances. The fingerprints similarity of 18 batches of samples was over 0.9, and the samples were classified into two groups. Six components were the main markers that cause differences in different batches of samples, including valepotriate, acevaltrate, isochlorogenic acid A, and some others. CONCLUSION: HPLC fingerprint combined with recognition of chemical pattern can reflect the intrinsic quality of Valeriana jatamansi, which may provide reference for the quality control and evaluation of Valeriana jatamansi.

Fingerprint study of Picrorhiza scrophulariiflora by UPLC coupled with chemometrics method / 中草药

Objective To establish the UPLC fingerprint for effective quality control and scientific evaluation of Picrorhiza scrophulariiflora. Methods The analysis was performed on Waters ACQUITY UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 μm), using acetonitrile-0.5% glacial acetic acid aqueous solution as mobile phase for gradient elution, with the flow rate at 0.3 mL/min, the column temperature at 32 ℃, and the detection wavelength at 295 nm. Total of 25 batches of P. scrophulariiflora and its adulterants were analyzed. Similarity evaluation combined with hierarchical clustering analysis (HCA) and principal components analysis (PCA) were used to evaluate the quality of herbs from different batches. Ultra-performance liquid chromatography- quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF/MS) was used for qualitative analysis in the positive and negative ion modes. Results There were significant differences in fingerprint chromatogram among P. scrophulariiflora and its adulterants. There were 16 common peaks in UPLC fingerprint of 22 batches of P. scrophulariiflora, and 12 peaks among which were carried out for chemical components identification with the similarity at 0.939-0.998. Twenty-two samples could be classified into three clusters. The PCA result was consistent with that of HCA. The four symbolic compounds in samples were verified by PLS-DA analysis, which identified that No.1, 12, 9 peaks were picroside I, picroside III, and scrophenoside C. Conclusion The establishment of UPLC fingerprint and the recognition of chemical pattern of P. scrophulariiflora can provide a more comprehensive reference for the quality control of herbs.

Comprehensive evaluation of Yiqi Fumai Injection quality by UPLC-Q-TOF- MSE fingerprint and chemical pattern recognition / 中草药

Objective To establish a UPLC-Q-TOF-MSE fingerprint of Yiqi Fumai Injection (YQFM) for providing reference for visual, easy and overall control of its quality. Methods The chromatographic separation was performed on a Waters Acquity UPLC HSS T3 (100 mm × 2.1 mm, 1.8 μm) column with the mobile phase consisting of acetonitrile and 0.1% formic acid for gradient elution. The flow rate was 0.3 mL/min, and the column temperature was 30 ℃. The capillary voltage was set at 2.5 kV. The nebulization gas was set to 800 L/h at 400 ℃, and the source temperature was 100 ℃. The BPC obtained with negative ion ESI mass spectra were selected for the fingerprint analysis. Similarity evaluation was used to evaluate the quality of YQFM from different batches. Based on the intensities of the ions for common peaks, HCA and PCA were performed using SPSS 19.0 and Simca-P software. Results The UPLC-Q-TOF-MSE fingerprint of YQFM was established by using 28 batches of sample and 18 common peaks were found, of which 15 mutual peaks from Ginseng Radix et Rhizoma rubra, three mutual peaks from Ophiopogonis Radix. Compared with the reference substances and references, 16 of the common peaks were identified and the similarity of 28 batches samples were over 0.970. 28 batches of YQFM could be divided into four grades when the sum of squared Euclidean distance is 5-10 in the result of HCA; PCA got seven principal components through dimension reduction and accumulative contribution rate reached 84.989%. By fitting the load factor model of the first principal component, ten markers greatly impacting on the quality were found. The comprehensive evaluation function of YQFM in different batches was constructed according to the principal component score. Among 28 batches of YQFM, the comprehensive score of S28 was the best, closely followed by S22, S11 and S9, while S14 and S13 was the worst. Conclusion The utilization of UPLC-Q-TOF-MSE fingerprint coupled with chemical pattern recognition could objectively and effectively assess the quality of YQFM, can provide a more comprehensive reference for the quality control of YQFM.

Flavonoids rather than alkaloids as the diagnostic constituents to distinguish Sophorae Flavescentis Radix from Sophorae Tonkinensis Radix et Rhizoma: an HPLC fingerprint study / 中国天然药物

Sophorae Flavescentis Radix (Sophora flavescens Ait., SFR) and Sophorae Tonkinensis Radix et Rhizoma (S. tonkinensis Gapnep., STR) are two commonly used traditional Chinese medicines from Sophora (Leguminosae) plants, which are believed to possess similar bioactive components with entirely different clinical applications. In order to find out the characteristic chemical constituents potentially leading to the unique medicinal properties claimed for each of the two closely related TCMs, an HPLC fingerprint method was developed for analyses of the alkaloid and flavonoid constituents of SFR and STR, respectively, which were further evaluated and compared through similarity calculation and hierarchical clustering analysis (HCA). The results from the present study showed that the alkaloid fingerprints of the two herbs were similar, with many components co-existing in both drugs and various batches of samples from different species being mixed together in the HCA dendrogram. However, their flavonoid constituents were totally different with specific fingerprints being yielded for each herb, and further HCA analysis showed that the tested samples could almost be clearly divided into two groups based on their origins of species. The results from the present study indicated that the flavonoid constituents could serve as the differentially diagnostic constituents of SFR and STR and might potentially attributed to their distinct therapeutic effects.

Analysis on multiple pharmaceutical ingredients and antioxidant capacities of Prunellae Spica based on multivariate statistical analysis / 中国中药杂志

Prunellae Spica is a perennial edible and medicinal plant, rich in antioxidant substances. Total flavonoids (TFC), Phenolics (TPC), triterpenoids (TSC), polysaccharides (PC) and their antioxidant capacities (by the FRAP, DPPH and ABTS⁺ methods) of ethyl acetate fraction, n-butanol fraction and other fractions of aqueous extract from Prunellae Spica were investigated in this study. Then the multivariate statistical method was adopted to analyze the relationship between the multiple pharmaceutical ingredients and antioxidant capacities of Prunellae Spica. The results showed that ethyl acetate fraction had relatively high concentration of TFC (0.61±0.10) g·g⁻¹DW, TPC (0.52±0.09) g·g⁻¹DW, and TSC (0.21±0.03) g·g⁻¹DW, with high scavenging capacity of DPPH (3.1±0.38) mmol·L⁻¹·g⁻¹DW and FRAP (2.56±0.35) mmol·L⁻¹·g⁻¹DW. Hierarchical clustering analysis (HCA) and principal component analysis (PCA) results indicated the information from chemical compositions and antioxidant capacity can represent the "differences" of different fractions. Canonical correlation analysis (CCorA) revealed a high positive correlation between the amounts of multiple chemical compositions and the antioxidant capacities (r=0.970 0), and the first canonical variate had been reached. Moreover, ABTS⁺ method showed a low response to the compositions of different fractions, so this method may not be suitable for evaluation of Prunellae Spica antioxidant capacities, while DPPH evaluation method was more suitable for TSC and TPC. The results of this study have important reference significance for the evaluation method on antioxidant activity of Prunellae Spica in the field of food or medicine as well as for the development of related extracts.

Flavonoids rather than alkaloids as the diagnostic constituents to distinguish Sophorae Flavescentis Radix from Sophorae Tonkinensis Radix et Rhizoma: an HPLC fingerprint study / 中国天然药物

Sophorae Flavescentis Radix (Sophora flavescens Ait., SFR) and Sophorae Tonkinensis Radix et Rhizoma (S. tonkinensis Gapnep., STR) are two commonly used traditional Chinese medicines from Sophora (Leguminosae) plants, which are believed to possess similar bioactive components with entirely different clinical applications. In order to find out the characteristic chemical constituents potentially leading to the unique medicinal properties claimed for each of the two closely related TCMs, an HPLC fingerprint method was developed for analyses of the alkaloid and flavonoid constituents of SFR and STR, respectively, which were further evaluated and compared through similarity calculation and hierarchical clustering analysis (HCA). The results from the present study showed that the alkaloid fingerprints of the two herbs were similar, with many components co-existing in both drugs and various batches of samples from different species being mixed together in the HCA dendrogram. However, their flavonoid constituents were totally different with specific fingerprints being yielded for each herb, and further HCA analysis showed that the tested samples could almost be clearly divided into two groups based on their origins of species. The results from the present study indicated that the flavonoid constituents could serve as the differentially diagnostic constituents of SFR and STR and might potentially attributed to their distinct therapeutic effects.

HPLC Fingerprint Analysis of Caulis of Chimonanthus nitens Combined with Chemometrics Method / 中国药学杂志

OBJECTIVE: To establish the high performance liquid chromatography (HPLC) fingerprint of the caulis of Chimonanthus nitens and evaluate the product quality by chemometrics analysis method. METHODS: The method was developed on an Amethyst C18-H column(4.6 mm×250 mm, 5 μm)by gradient elution with acetonitrile-water (containing 0.1% formic acid) as mobile phase at a flow rate of 0.8 mLmin-1. The column temperature was maintained at 28℃, and the detection wavelength was set at 254 nm. The main characteristic peaks was identified by comparing the retention time and UV absorption characteristics. Then 10 batches of the caulis of Chimonanthus nitens were evaluated by similarity assay, HCA, and PCA. RESULTS: The HPLC fingerprint of the caulis of Chimonanthus nitens was established and three main peaks were identified. The similarity of 10 batches of the caulis of Chimonanthus nitens was about 0.978 0 to 0.991 9. CONCLUSION: The established method can be used for the quality control of the caulis of Chimonanthus nitens.

Study on fingerprint analysis of Xihuang Pills by HPLC-ELSD / 中草药

Objective: To establish the quality evaluation of fingerprints of Xihuang Pills by HPLC-ELSD. Methods: The chromatography conditions were defined as waters C18 column (250 mm × 4.6 mm, 5 μm); Mobile phase was methanol-0.5% acetic acid, gradient elution; temperature of column was set at 30℃. The ELSD conditions were as follows: the temperature of drift tube was 45℃, the gas speed was 1.5 L/min. Ten batches of Xihuang Pills samples were analyzed for similarity analysis (SA), hierarchical clustering analysis (HCA) and principle component analysis (PCA). Results: The chromatographic fingerprint was completed with 25 recognizable peaks, and the samples with great differences and the compounds with greater impact on the quality were obtained through HCA and PCA. Conclusion: HPLC fingerprint combining with pattern recognition could reflect the intrinsic quality to provide a scientific basis for the quality control of Xihuang Pills.

Content Analysis and Comparative Study of Amino Acids in Ejiao from 18 Manufacturers / 中国药房

OBJECTIVE:To provide reference for quality evaluation of Ejiao. METHODS:The contents of 17 amino acids in 18 batches of Ejiao from 18 manufacturers were analyzed by automatic amino acid analyzer;the content data was processed by prin-cipal component analysis (PCA) and hierarchical clustering analysis (HCA),in order to understand the relationship among these amino acids and categorize the Ejiao products. RESULTS:Except for a batch of Ejiao,the other 17 batches contained 17 various amino acids,including 7 necessary amino acids(Thr,Val,Met,Ile,Leu,Phe and Lys)for human being,especially the contents of Gly and Pro were higher. The total content of amino acids was arranged from 66.1% to 82.0%,showing great difference. Four main components were extracted by PCA,the cumulative contribution of which was 89.578%,and accordingly it may be consid-ered that Pro,Ala,Glu,Asp,Leu,Ile and Thr were characteristic amino acids of Ejiao products. In hierarchical clustering analy-sis,3 tree figures of commercial Ejiao product were achieved and Euclidean distance was varied among 0-25. CONCLUSIONS:There is great difference in total control of amino acid in commercial Ejiao product;Ejiao products from various manufacturers dif-fer in their amino acid contents. Ejiao is rich in collagen,and therefore,using amino acid determination would be helpful to moni-tor its quality.

Fingerprint of Sarcandrae Herba by UPLC coupled with chemometrics methods / 中草药

To establish the UPLC fingerprint for effective quality control of Sarcandrae Herba. The fingerprint of Sarcandrae Herba was developed with ultra-performance liquid chromatography, and the Acquity UPLC HSS T3 column (50 mm × 2.1 mm, 1.8 μm) was used in the gradient elution with a mobile phase of acetonitrile-water (both containing 0.1% formic acid): The flow rate was 0.5 mL/min, the column temperature was 35℃, and the detection wavelength was at 330 nm. Similarity evaluation combined with hierarchical clustering analysis (HCA) and principal components analysis (PCA) were used to evaluate the quality of herbs from different areas. The fingerprint of Sarcandrae Herba was established with good precision, reproducibility, and stability obtaining within 23 min, and nine peaks in the fingerprint were designed. Eighteen samples could be classified into two clusters. The PCA result was consistent with the HCA. A brief list of five differential compounds is presented, including rosmarinic acid, newchlorogenic acid, and so on. The establishment of UPLC fingerprint of Sarcandrae Herba and the application of chemical pattern recognition can provide a more comprehensive reference for the quality control of herbs.

UPLC fingerprint of Paeoniae Alba Radix from different regions / 中草药

Objective: To establish a UPLC fingerprint method of Paeoniae Alba Radix, and provide comprehensive evaluation of their quality in different regions. Methods: The UPLC chromatographic column was Acquity UPLC® HSS T3 (100 mm × 2.1 mm, 1.8 μm). The mobile phase was acetonitrile-0.05% phosphoric acid water with gradient elution. The detection wavelength was 230 nm and column temperature was 30 ℃ with the flow rate of 0.4 mL/min. Similarity analysis, hierarchical clustering analysis, and principal component analysis were undertaken to study 23 sets of UPLC fingerprints of Paeoniae Alba Radix. Results: A specific UPLC fingerprint of Paeoniae Alba Radix was established and eight common peaks were designated. The results showed that the qualities of the 23 sets of Paeoniae Alba Radix were not stable and the samples collected from same region and different regions both had certain differences. Conclusion: UPLC fingerprint is an available and convenient method which can be used to access the quality of Paeoniae Alba Radix rapidly.

HPLC fingerprint and chemical pattern recognization of Kudiezi injection / 中国药学杂志

OBJECTIVE: To develop a method to establish the fingerprint of traditional Chinese medicines Kudiezi injection by HPLC, and Assess the quality of Kudiezi injection by chemical pattern recognization techniques. METHODS: HPLC method was used to establish Kudiezi injection standard fingerprint consisting of 15 common peaks and 14 compounds were identified. The fingerprint was further analyzed by chemometric methods including similarity analysis (SA), hierarchical clustering analysis (HCA), principal component analysis (PCA). RESULTS: The similarity of 14 batches of Kudiezi injection was greater than 0.94. The samples were clearly classified into two groups by using PCA and HCA, respectively, and the results of classification were consistent. CONCLUSION: The developed method of chromatographic fingerprint is noval, simple and reliable. The results indicate that the chemical pattern recognization is suitable for the analysis of fingerprint data, which can be applied as one measure for the quality evaluation of Kudiezi injection.

Evaluation preparation technology of Xiaochaihu granules using fingerprint-peak pattern matching / 药物分析学报

An approach was proposed to evaluate preparation technology by means of fingerprint-peak matching technology of high performance liquid chromatography with diode array detector （HPLC-DAD）. Similarity and hierarchical clustering analysis （HCA） were applied to identify the 15 batches of Xiaochaihu granules from different manufacturers and our laboratory, and peak pattern matching between the composite formulae and Radix Bupleuri Chinensis, which was one of the main ingredients of Xiaochaihu granules, was utilized to evaluate the preparation technology of Xiaochaihu granules via the indexes of the relative deviation of retention time （RT） and UV spectrum feature similarity of their corresponding peaks. Eleven matching peaks were found between Xiaochaihu granules and Radix Bupleuri Chinensis. However, the saikosaponin A and saikosaponin D, which are the important active components in Radix Bupleuri Chinensis, were not found in Xiaochaihu granules from any manufacturers. The peak areas of 11 characteristic peaks of Xiaochaihu granules samples formed a matrix of 11 × 15. The result of HCA showed that Xiaochaihu granules samples were divided into four kinds of category. Xiaochaihu granules samples from the same manufacturer were basically clustered of the same category. The results suggested that the saikosaponin A and saikosaponin D are prone to structural transformation under the condition of decoction and in the presence of the organic acidic components. These active components, existing in raw herb, might transform to a series of non-active secondary saikosaponin due to unfavourable preparation technology. So the conventional decoction-based preparation technology of Xiaochaihu granules might greatly affect its quality and therapeutic effectiveness. This study demonstrates that fingerprint-peak matching technology can not only be used for quality control of this composite formulae, but also provide some guidance for preparation technology of Xiaochaihu granules.

Evaluation preparation technology of Xiaochaihu granules using fingerprint-peak pattern matching / 药物分析学报

An approach was proposed to evaluate preparation technology by means of fingerprint-peak matching technology of high performance liquid chromatography with diode array detector (HPLC-DAD).Similarity and hierarchical clustering analysis (HCA) were applied to identify the 15 batches of Xiaochaihu granules from different manufacturers and our laboratory,and peak pattern matching between the composite formulae and Radix Bupleuri Chinensis,which was one of the main ingredients of Xiaochaihu granules,was utilized to evaluate the preparation technology of Xiaochaihu granules via the indexes of the relative deviation of retention time (RT) and UV spectrum feature similarity of their corresponding peaks.Eleven matching peaks were found between Xiaochalhu granules and Radix Bupleuri Chinensis.However,the saikosaponin A and saikosaponin D,which are the important active components in Radix Bupleuri Chinensis,were not found in Xiaochaihu granules from any manufacturers.The peak areas of 11 characteristic peaks of Xiaochaihu granules samples formed a matrix of 11 × 15.The result of HCA showed that Xiaochaihu granules samples were divided into four kinds of category.Xiaochaihu granules samples from the same manufacturer were basically clustered of the same category.The results suggested that the saikosaponin A and saikosaponin D are prone to structural transformation under the condition of decoction and in the presence of the organic acidic components.These active components,existing in raw herb,might transform to a series of non-active secondary saikosaponin due to unfavourable preparation technology.So the conventional decoction-based preparation technology of Xiaochaihu granules might greatly affect its quality and therapeutic effectiveness. This study demonstrates that fingerprint-peak matching technology can not only be used for quality control of this composite formulae,but also provide some guidance for preparation technology of Xiaochaihu granules.

Classification of A Medically Important Group of Gamasid Mites by Numerical Taxonomy in Yunnan, China / 热带医学杂志

Objective To determine the taxonomic position of medically important gamasid mites and to compare numerical taxonomy with the traditional systematics in the classification of gamasid mites. Methods Based on hierarchical cluster analysis, numerical taxonomy was applied to study fifty-seven species of medically important gamasid mites. Results The results of cluster analysis based on squared euclidean distance showed that Hirstionyssus Fonseca and Echinonyssus Hirst should be separated from Laelapidae Berlese and formed an independent family. The taxonomic position of the remaining species remains the same as those in the traditional systematics. Conclusion Numerical taxonomy can objectively reflect the taxonomic position of the medically important gamasid mites. The result of classification by numerical taxonomy is consistent with traditional systematics for gamasid mites.