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OBJECTIVE To establish the characteristic chromatogram of Chaenomeles sinensis， determine the contents of rutin， hyperin and quercitrin， and to identify C. sinensis and C. speciosa. METHODS HPLC method was performed on Agilent 5 TC-C18 column， with acetonitrile-0.2% formic acid solution as the mobile phase for gradient elution， at the flow rate of 1.0 mL/min. The column temperature was 30 ℃ . The detection wavelength was 330 nm in characteristic chromatogram and 350 nm in content determination. The characteristic chromatogram of C. sinensis was established and similarity was evaluated by the Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 edition）. Hierarchical cluster analysis of 15 batches of C. sinensis （S1-S15） was performed by using SPSS 23.0 software. The contents of 3 flavones in 15 batches of C. sinensis and 7 batches of C. speciosa （S16-S22） were determined， while their characteristic chromatograms were compared. RESULTS The similarities of the characteristic chromatogram for 15 batches of C. sinensis ranged from 0.783 to 0.969， and 11 characteristic peaks were confirmed. Four constituents were identified as chlorogenic acid， rutin， hyperin and quercitrin. The medicinal materials in 15 batches of C. sinensis could be divided into 2 categories： S5-S8 were one category， and the others belonged to one category. The characteristic chromatogram of C. sinensis was obviously different from C. speciosa. The contents of rutin， hyperin and quercitrin in 15 batches of C. sinensis were 48.99-294.45， 3.49-102.55， 31.98-149.49 μg/g， respectively. The content of rutin in C. speciosa was lower than that in C. sinensis. None of hyperin （except for S20） and quercitrin were detected in C. speciosa. CONCLUSIONS The characteristic chromatogram and the method for content determination of 3 flavones in C. sinensis are established successfully and can be used for the quality control of C. sinensis and its identification from C. speciosa.
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Objective:To comprehensively evaluated the quality of Sargentodoxae Caulis from different habitats with a combination of indexes and characteristic chromatogram method from Chinese Pharmcopoeia (Edition 2020). Methods:The contents of water content, total ash, ethanolic extract, sulfur dioxide residue, heavy metals and harmful elements, total phenols, chlorogenic acid, salidroside and characteristic chromatogram of 17 batches of Sargentodoxae Caulis were determined. The quality of Sargentodoxae Caulis was comprehensively evaluated by combining chemical pattern recognition method. Results:The water content, total ash content, extracts, and content determination of 17 batches of Sargentodoxae Caulis from different habitats complyed with the provisions of the Chinese Pharmcopoeia (Edition 2020). There were differences in the contents of extracts, chlorogenic acid, and salidroside, among which the content of Anhui origin was higher. A total of 8 common peaks were identified from the 17 batches samples. Conclusion:Comprehensive evaluation of multiple indicators can demonstrate the quality of Sargentodoxae Caulis more correctly, and shows that the quality of Sargentodoxae Caulis from different habitats is different. The quality of Sargentodoxae Caulis from Anhui is better than that from other habitats.
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OBJECTIVE To compare characteristic chromatogram and the contents of multiple indicator components of Morus alba decoction powder and decoction at different decoction time， and to provide experimental basis for the development of M. alba decoction. METHODS Taking decoction powder and decoction at different decoction time as subject， HPLC characteristic chromatogram of 2 kinds of samples were established with Similarity Evaluation Software System of TCM Chromatographic Fingerprint （2012 version）， and similarity evaluation was performed. The contents of mulberroside A， geniposide， berberine， baicalin， quercetin and luteolin in decoction powder and decoction were determined by HPLC. The contents of each indicator component and the change of total content were as the evaluation indexes to compare the difference between the two substances during decoction. RESULTS The similarities of characteristic chromatogram of the two substances ranged from 0.943 to 1.000 and 0.975 to 0.998 at different decoction time， respectively. Six indicator components of the decoction powder dissolved faster and had higher contents. The contents of each indicator component in the decoction powder when decocting at 20 minutes was 1.1-1.5 times of the decoction when decocting at 50 min， and the total content in the decoction powder was 1.2 times of the decoction. CONCLUSIONS Compared with decoction， M. alba decoction powder has the advantages of shortening the decoction time and saving traditional Chinese medicine resources. The results of this study lay a research foundation for “Zungu” to develop its preparation.
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Objectives To provide scientific basis for the quality control of different species of Baishouwu by establishing the HPLC fingerprint of domestic of Baishouwu and determining the main active components of acetophenones. Methods HPLC-DAD method was used to determine the HPLC fingerprints of domestic of Baishouwu. Then, the content of 4 kinds of acetophenones in Baishouwu was determined. The column was Diamonsil C18（250mm×4.6mm, 5μm）with the mobile phase of methanol and 0.1% phosphoric acid at a flow rate of 1.0 ml/min. The detection wavelength of p-Hydroxyl acetophenone, baishouwu benzophenone, 2',4'-Dihydroxy acetophenone was set at 260 nm and 2',5'-Dihydroxy acetophenone at 280 nm respectively. Results The similarity and cluster analysis in HPLC fingerprint showed that the constituents were significantly different among C. bungei, C. auriculatum and C. wilfordii. The content of total acetophenones in C. wilfordii was significantly higher than that in other localities of C. auriculatum and C. bungei. Conclusions Acetophenone could be used as the evaluation index to evaluate the quality of Baishouwu in different origins. The content of total acetophenone in C. wilfordii is the highest, which could be used as the best quality resource of Baishouwu.
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OBJECTIVE To establish HPLC characteristic chro matogram of Jianpi yifei biyan prescription standard decoction ， to select the quality control index components and determine their contents. METHODS HPLC method combined with Similarity Evaluation System of TCM Chromatographic Fingerprint （2004 edition）were used to establish the characteristic chromatogram of 10 batches of Jianpi yifei biyan prescription standard decoction ；the similarity evaluation and common peaks identification were also carried out. Using common peak area of characteristic chromatogram as variables ，SPSS 26.0 software and SIMCA 14.1 software were used to perfor m cluster analysis （CA），principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA）；differential components with variable important i n pro jection（VIP）value greater than 1.5 were screened；the contents of cimifugin and differential components were determined by the same method. RESULTS A total of 24 common characteristic peaks were identified ， and the similarities of 10 batches of samples were higher than 0.960；eight characteristic peaks were identified by comparison with reference substance. CA and PCA results revealed that the samples were classified into 3 categories.OPLS-DA analysis showed that 3 components with VIP value greater than 1.5， which were prim-O-glucosylcimifugin （peak 2），calycosin 7-O-β-D-glucopyranoside （peak 4） and 5-O-methylvisammioside （peak 6） in descending order. The linear ranges of prim- O- glucosylcimifugin，calycosin 7-O-β-D-glucopyranoside，cimifugin and 5-O-methylvisammioside were 0.010 7-0.213 0，0.007 8- 0.156 0，0.008 0-0.160 0，0.009 8-0.195 0 μg（r＞0.999），respectively. RSD values of precision ，repeatability and stability tests （24 h） were all less than 2％. Average recoveries were 105.98％（RSD＝1.75％，n＝6），98.06％（RSD＝3.87％，n＝6），96.38％（RSD＝ 4.03％ ，n＝6） and 104.17％（RSD＝1.27％ ，n＝6）. The contents of the above 4 components in 10 batches of samples were 12.12-18.87，3.86-6.40，3.10-4.27 and 11.17-15.79 μ g/mL，respectively. CONCLUSIONS The established HPLC characteristic chromatographic method is stable and feasible ，it can be used for the quality control of Jianpi yifei biyan prescription standard decoction. Prim- O-glucosylcimifugin，calycosin 7-O-β-D-glucopyranoside，cimifugin and 5-O-methylvisammioside can be used as the index components for quality control of the standard decoction.
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The methods for determining the characteristic chromatogram and index components content of Xuanfu Daizhe Decoction were established to provide a scientific basis for the quality evaluation of substance benchmarks and preparations. Eighteen batches of Xuanfu Daizhe Decoction were prepared with the decoction pieces of different batches and of the same batch were prepared respectively, and the HPLC characteristic chromatograms of these samples were established. The similarities of the chromatographic fingerprints were analyzed. With liquiritin, glycyrrhizic acid, 6-gingerol, ginsenoside Rg_1, and ginsenoside Re as index components, the high performance liquid chromatography was established for content determination with no more than 70%-130% of the mass average as the limit. The results showed that there were 19 characteristic peaks corresponding to the characteristic chromatograms of 18 batches of Xuanfu Daizhe Decoction, including 8 peaks representing liquiritin, 1,5-O-dicaffeoylqunic acid, ginsenoside Rg_1, ginsenoside Re, 1-O-acetyl britannilactone, ginsenoside Rb_1, glycyrrhizic acid, and 6-gingerol, and the fingerprint similarity was greater than 0.97. The contents of liquiritin, glycyrrhizic acid, 6-gingerol, and ginsenosides Rg_1 + Re in the prepared Xuanfu Daizhe Decoction samples were 0.53%-0.86%, 0.61%-1.2%, 0.023%-0.068%, and 0.33%-0.66%, respectively. Except for several batches, most batches of Xuanfu Daizhe Decoction showed stable contents of index components, with no discrete values. The characteristic chromatograms and index components content characterized the information of Inulae Flos, Ginseng Radix et Rhizoma, Glycyrrhizae Radix et Rhizoma, and Zingiberis Rhizoma Recens in Xuanfu Daizhe Decoction. This study provides a scientific basis for the further research on the key chemical properties of substance benchmark and preparations of Xuanfu Daizhe Decoction.
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Benchmarking , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/chemistry , Ginsenosides/analysis , Glycyrrhizic Acid/analysis , Quality ControlABSTRACT
The high performance liquid chromatography(HPLC) characteristic chromatogram of Xiaoer Ganmaoning Oral Liquid(oral liquid for short) was established. The medicinal materials corresponding to characteristic peaks, their index components and ranges of similarity with the reference chromatograms were clarified. The similarity between the characteristic chromatograms of 10 batches of the oral liquid and the reference chromatogram was higher than 0.994. Eighteen characteristic peaks were identified, which were derived from different medicinal materials including Scutellariae Radix, Arctii Fructus, Lonicerae Japonicae Flos, Gardeniae Fructus and Forsythiae Fructus. Further, 11 characteristic peaks were assigned by the comparison with reference substances as chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A, isochlorogenic acid C, baicalin, baicalein, wogonin, scutellarin, forsythiaside A and arctiin. Also, the characteristic chromatogram of precipitate in the oral liquid was established, and the similarity between characteristic chromatograms of 10 batches of the precipitate and the reference chromatogram was higher than 0.940. The 14 characteristic peaks originating from the precipitate and those from the oral liquid were consistent in retention time, and the content of all index components in the precipitate was lower than 5% of that in the oral liquid. Moreover, the stability of precipitate during the accelerated stability test was explored with filtration and Matlab-based image sensory evaluation. The precipitate mass and precipitation degree both increased over the stability test duration significantly. The stability of the oral liquid was used as a model system in this study to establish the integrated quality control system which related to medicinal materials, preparations and precipitate with HPLC characteristic chromatograms and image sensory evaluation, which lays a foundation for the exploration of the quantity value transfer of the oral liquid.
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Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal/chemistry , Quality Control , Scutellaria baicalensis/chemistryABSTRACT
In light of related methods in Chinese Pharmacopoeia(2020 edition), this study established the quality standard for Lobeliae Chinensis Herba. The TLC identification method was established with silica gel GF_(254) thin layer plate, diosmin standard, linarin standard, and the reference material of Lobeliae Chinensis Herba. The loss on drying, total ash, acid-insoluble ash, and ethanol-soluble extracts of 18 batches of Lobeliae Chinensis Herba samples were determined according to the general principles in Chinese Pharmacopoeia. Then, HPLC was adopted in the establishment of characteristic chromatogram and content determination. The results showed that the established method can achieve good separation for diosmin, linarin, and lobetyolin. Based on the results of detection for 18 batches of Lobeliae Chinensis Herba samples, the draft quality standard was established, which was expected to provide reference for the revision of this medicinal herb in Chinese Pharmacopoeia.
Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/standards , Lobelia/chemistry , Plants, Medicinal/chemistryABSTRACT
OBJECTIVE: To establish the characteristic chromatogram of Baphicacanthis Cusiae Rhizoma et Radix and its decoction pieces by HPLC for the identification of authentic and counterfeit products. METHODS: High performance liquid chromatography (HPLC) was used with Agilent Zorbax C18(4.6 mm×250 mm, 5 μm). The mobile phase was acetonitrile-water with gradient elution. The detector was a secondary tube array (DAD). The column temperature was maintained at 35 ℃, the flow rate was 1.0 mL•min-1, and the injection volume was 10 μL. RESULTS: Fifteen batches of genuine crude drug and twelve batches of genuine decoction pieces were determined. Five common peaks were found, among which three peaks were 2-benzoxazolinone, indigo and indirubin. CONCLUSION: The established characteristic chromatogram of Baphicacanthis Cusiae Rhizoma et Radix can effectively distinguish the authentic from the counterfeit. The methodological demonstration shows that the method is accurate, stable and reproducible.
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Objective: To establish and analyze the HPLC characteristic chromatogram for water extracts of Arisaematis Rhizoma (AR) and its processed products, Arisaematis Rhizoma Preparatum (ARP) and Arisaema Cum Bile (ACB). The research provided reliable method and scientific basis for their quality control. Methods: The separation was performed on an Agilent C18 (200 mm × 4.6 mm, 5 μm) column with gradient elution of 0.2% acetic acid water and 0.2% acetic acid acetonitrile. The similarity was analyzed with software “Similarity Evaluation System for Chromatographic Fingerprint of TCMs (Version 2012)”. The cluster analysis was performed by SPSS 23.0. The principal component analysis was performed by SIMCA 14.1. Results: HPLC characteristic chromatogram for water extracts of AR, ARP, and ACB were established. There were 19, 20 and 13 common peaks in AR, ARP and ACB, respectively. A total of eight characteristic peaks were identified as F1 (xanthine), F3 (uracil), F4 (hypoxanthine), F5 (uridine), F6 (guanosine), F7 (adenosine), F14 (schaftoside), and F16 (isoschaftoside), respectively. The intragroup similarities of AR, ARP, and ACB were all above 0.8 and each was clustered into one type, and the intergroup similarities among AR and its processed products were all below 0.4. The main components of AR were F16 (isoschaftoside), F14 (schaftoside), F17and F15. The main components of ARP were F16 (isoschaftoside) and F1 (xanthine). The main components of ACB were F3 (uracil), F2, F5 (uridine) and F1 (xanthine). Conclusion: The method can effectively identify AR, ARP and ACB, and provide a scientific basis for their quality control.
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Objective: To compare the chemical compositions of Scutellariae Radix (SR) before and after wine-frying, and provide a reference for establishing a comprehensive quality evaluation method for SR decoction pieces. Methods: Characteristic chromatogram of SR and wine-processed Scutellariae Radix (wpSR) were established using HPLC, the control characteristic chromatogram maps were generated, the common peaks were calibrated and the similarity was evaluated. Chemical compositions of SR and wpSR were identified by UPLC-Q-TOF/MS. The content profiles of 11 flavonoids (baicalin, baicalein, wogonoside, wogonin, oroxylin A, oroxyloside, scutellarin, apigenin, hispidulin, luteoloside and chrysin) of SR and wpSR was determined by UPLC-TQMS, and PCA was used to analysis the content of 11 flavonoids. Results: The characteristic chromatogram of SR and wpSR were established, and nine common peaks were calibrated. The similarity of two pieces were all greater than 0.947. Through the analysis of the multistage tandem mass spectrometry, retention time matching combined with the software of database search and literatures, 50 compositions were found and 44 compositions were identified in two pieces. The quantitative results showed that the content of flavonoid glycosides such as baicalin and wogonoside decreased slightly, while the content of flavonoid aglycones such as baicalein and wogonin increased slightly. After multivariate statistical analysis, two pieces were divided into two types. The differences of the content of baicalin, wogonoside and oroxyloside may be the main factors causing the change of chemical compositions of wine-frying of SR. Conclusion: Chemical profiles did not change after wine-frying of SR, but the content profiles of some compositions were changed. The established method can provide reproducible, efficient, as well as accurate analysis for Chinese medicinal materials. And it should be an eligible tool for the quality evaluation of SR.
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Objective: To establish a method for the characteristic chromatogram and determination of indicative components of salt-fried Phellodendri Chinensis Cortex (sPCC) and Yihuang Decoction (YHD). Methods: An HPLC was established for characteristic chromatogram analysis and determination of four indicative components (phellodendrine, 4-O-feruloylquinic acid, 5-O- feruloylquinic acid and berberine) of sPCC and YHD. Results: The characteristic chromatogram of sPCC and YHD were established by HPLC from 10 batches. Among five common peaks, four indicative components (phellodendrine, 4-O-feruloylquinic acid, 5-O-feruloylquinic acid and berberine) were identified and determined. The mass fraction of phellodendrine, 4-O-feruloylquinic acid, 5-O-feruloylquinic acid and berberine in sPCC ranged from 0.841%-1.314%, 0.358%-1.841%, 2.495%-5.498%, and 4.259%-7.007%, respectively; The mass concentration of phellodendrine, 4-O-feruloylquinic acid, 5-O-feruloylquinic acid and berberine in YHD ranged from 69.71-117.80, 107.85-165.79, 252.96-348.20, and 213.61-361.45 μg/mL, respectively; and the mass fraction of phellodendrine, 4-O-feruloylquinic acid, 5-O-feruloylquinic acid and berberine converted from YHD based on the mass of sPCC ranged from 0.509%-0.865%, 0.788%-1.212%, 1.849%-2.545%, and 1.561%-2.655%, respectively. Conclusion: The established method not only provides more complete reference and basis for the quality control, but also provides a certain basis for the research of effect substance basis of sPCC and YHD.
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Objective: A method for identification of root cortex and woody core of Morindae Officinalis Radix (MOR) was established based on Ultra Performance Liquid Chromatography (UPLC) characteristic chromatogram and chemical pattern recognition technique. Methods: Using UPLC technique, the characteristic chromatograms of iridoids and oligosaccharides of root cortex and woody core of MOR were established, combined with the similarity analysis, variance analysis, cluster analysis, principal component analysis (PCA) methods for chemical pattern recognition research. Results: The UPLC characteristic chromatograms of iridoids and oligosaccharides of different parts of MOR were established, and 12 and 20 common characteristic peaks were confirmed, respectively. The UPLC characteristic chromatograms of root cortex and woody core of MOR were obviously different. Conclusion: UPLC characteristic chromatograms of iridoids and oligosaccharides of MOR combined with chemical pattern recognition analysis method can reflect the difference of root cortex and woody core of MOR integrally, comprehensively and truly, which provides more sufficient basis for the necessity of removing woody core from MOR.
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Objective: A new table of standards of commodity classes of Saposhnikovia Radix is formed through literature market and local area survey. To explore the scientific nature and practicability of new standards of commodity classes of Saposhnikovia Radix, according to the appearance and index components related to quality of Saposhnikovia Radix, through statistical analysis and comparison of characteristic spectrum. Methods: Samples of different specifications and grades were collected from Northeast China, Inner Mongolia, Hebei, and the other main production areas, mainstream medicine market. Measuring samples appearance, extractum, characteristic spectrum of four chromones and analyzing data and discussing the relationship between standards of commodity classes and the appearance and index components to provide datas and theoretical support for the new table. Results: According to appearance character, the standards of commodity classes of Saposhnikovia Radix divided into three specifications of wild, imitates wild, cultivated were science based. At the same time, it is suggested to increase the content of the cimifugin as the index to distinguish between wild and cultivated Saposhnikovia Radix. Conclusion: There were significant differences in appearance and index components between different standards of Saposhnikovia Radix. Whether different classes of Saposhnikovia Radix only depend on size remains to be discussed.
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Objective: In order to provide a scientific basis for the quality control of Kunxian Capsules (KC), HPLC characteristics chromatogram combined with multi-components determination was established. Methods: The analysis was performed on Agilent Zorbax SB-C18 column (250 mm × 4.6 mm, 5 μm), using acetonitrile and 0.1% phosphoric acid solution as the mobile phase at a flow rate of 0.8 mL/min for gradient elution, the column temperature was 33 ℃, and the detection wavelength was 270 nm. The fingerprints of 15 batches of KC were established and evaluated by the similarity evaluation system of TCM (2012A version), hierarchical cluster analysis and discriminant analysis of partial least squares. Furthermore, the content of hyperoside, epimedin A, epimedin B, epimedin C, icariin and baohuoside Ⅰ were determined. Results: The HPLC fingerprint with 21 common peaks of KC was established, and the similarities of samples were over 0.9. The linearity relationships separated with hyperoside, epimedin A, epimedin B, epimedin C, icariin and baohuoside Ⅰ were good, and the contents of the above-mentioned components in 15 batches of preparations were 2.817-7.527, 7.287-9.103, 8.730-18.675, 33.377-70.371, 35.297-50.291 and 4.059-9.079 mg/g, respectively. Conclusion: The combination methods of HPLC characteristic chromatograms and simultaneous determinations of multiple components are rapid, simple and reproducible, which can provide methodological reference for the quality control of KC.
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This study aimed to establish the HPLC characteristic chromatogram and content determination method for index components with the primary standard substances of the classical prescription Mahuang Decoction, and to provide data basis for the establishment of its quality standard and the development and utilization of compound preparations. First, HPLC was used to establish the material reference chromatograms of Mahuang Decoction, and 15 batches of standard samples of Mahuang Decoction were determined. Their similarity was calculated by the median method. Secondly, the content of the standard substances was determined and a simplecontent determination method was established by HPLC. Relevant methodology was investigated, and the extraction ratio, index component transfer rate and moisture content of 15 batches of primary standard samples were calculated. The results showed that the two sets of HPLC methods had their own characteristics. The six chromatographic peaks identified from the 10 common peaks in the former characteristic chromatogram covered all the herbal medicines in the standard substances, which can better indicate the quality characteristics of the standard substances of Mahuang Decoction. The latter method(content determination method) was simple and practical, so it was suitable for establishing the quality standard of its compound preparation. Two sets of methods were jointly used to evaluate the quality of 15 batches of Mahuang Decoction. The results were as follows: the similarity of 15 batches of samples was greater than 0.90; the average extraction ratio was 18.1%; the average moisture content was 9.7%; the average content and transfer rate of the standard ingredients ephedrine hydrochloride and total pseudoephedrine hydrochloride were 2.3% and 26.7% respectively, and the average content and transfer rate of amygdalin were 2.2% and 48.3% respectively. None of the data showed dispersion(beyond 70%-130% of the mean value), which met the application data requirements for the substance standards of ancient classical Chinese herbal compound preparations(draft for comments). Based on the above research, the primary substance quality standard of Mahuang Decoction was established in order to provide reference for the development and research of the compound preparation of Mahuang Decoction.
Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Ephedra sinica , Prescriptions , Quality Control , Reference StandardsABSTRACT
Objective: To establish HPLC and multiple components determination method of Sarcandrae Herba Dispensing Granules (SHDG), in order to compare the difference of the quality in various SHDG samples and provide an effective method to ensure the quality of SHDG. Methods: HPLC-UV method was used to establish the characteristic chromatogram of SHDG, and acetonitrile-0.2% formic acid solution was used as the mobile phase with the gradient elution. The common peaks were identified by comparison with the reference standards and HPLC-Q/TOF. At the same time, the method for simultaneous determination of chlorogenic acid, isofraxidin, and rosmarinic acid was established with the same approach. Chemometrics software Chempattern was employed to analyze the data. Through cluster analysis and principal component analysis, different preparations from different manufacturers and different batches from the same manufacturer were classified, and the main components causing the differences were clarified. Results: The SHDG fingerprint was established to confirm and identify seven characteristic peaks, namely, neochlorgenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isofraxidin, rosmarinic acid-4-O-β-D-glucoside, and rosmarinic acid. The main chromatographic peaks of SHDG can be completely separated within 55 min. There was a good linear relationship between the peak area and concentration of chlorogenic acid, isofraxidin, and rosmarinic acid. The average recovery rates were 98.92% (RSD 1.54%), 98.20% (RSD 1.12%), and 99.58% (RSD 1.12%), respectively. The chlorogenic acid content of 18 batches of samples was 0.33-1.39 mg/g, the content of isocyanidine was 1.31-2.74 mg/g, and the content of rosmarinic acid was 1.11-4.54 mg/g. The similarity between 18 batches of samples and the common mode was 0.688-0.992. There were certain differences in the content of chlorogenic acid, isofraxidin, rosmarinic acid in the SHDG of various batches and manufacturers. Conclusion: The proposed specific HPLC characteristic chromatogram and quantitation method of three components for SHDG offered more comprehensive reference for quality control of the crude drug.
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Objective: To optimize purification process of total alkaloid extract of Berberis dictyophylla cortex by macroporous resin,and to establish its quality standard. Method: Acid dye colorimetry was used to investigate the purification process of total alkaloid extract of B. dictyophylla cortex,the process parameters included concentration of sample solution,speed of sampling,diameter-height ratio of resin column,water washing amount,concentration and dosage of eluent,flow rate of elution,etc.In order to determine the optimum process,HPLC was employed to determine the contents of four alkaloids(magnoflorine,jatrorrhizine hydrochloride,palmatine hydrochloride,and berberine hydrochloride) with mobile phase of acetonitrile-0.1% phosphoric acid aqueous solution for gradient elution and detection wavelength at 270 nm.After being purified,quality standard of total alkaloid extract of B. dictyophylla cortex was investigated according to the requirements in the 2015 edition of Chinese Pharmacopoeia. Result: Optimal purification conditions were as following:10 g of HPD100 macroporous adsorption resin with a column diameter-height ratio of 1:8,sampling solution concentration of 11 g·L-1,the loading flow rate of 1 mL·min-1,sampling solution volume of 50 mL,washed with 4 BV of water(1 BV=15 mL) and added 9 BV of 30% ethanol,after being purified,the transfer rate of total alkaloids was>80%,and its purity was>65%.The quality standard of total alkaloid extract of B. dictyophylla cortex was established,there were 19 common peaks in the characteristic chromatogram,and the overall similarity was>0.99. Conclusion: This optimized purification process is stable and feasible, and the established quality standard is controllable.
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Objective: To establish HPLC characteristic chromatogram of standard pieces of Polygoni Multiflori Radix, and compare with the HPLC characteristic chromatograms of original pieces, raw materials and control materials of Polygoni Multiflori Radix. Method: HPLC analysis was carried out on a Waters BEH-C18 column (4.6 mm×250 mm, 5 μm) with the mobile phase of methanol(A)-0.1%phosphoric acid(B) for gradient elution (0-20 min, 15%-30%A; 20-35 min, 30%-40%A; 35-55 min, 40%-75%A; 55-75 min, 75%-100%A) at a flow rate of 0.8 mL·min-1. The detection wavelength was set at 270 nm and the column temperature was maintained at 30℃. The injection volume was 10 μL. Moreover, similarity and cluster analysis of HPLC characteristic chromatograms of four samples of Polygoni Multiflori Radix were performed. Result: HPLC characteristic chromatogram of standard pieces of Polygoni Multiflori Radix composed of seven peaks was established. The similarity was 0.999 between characteristic chromatograms of standard pieces and original pieces, which was better than similarities among characteristic chromatograms of raw materials, control materials and original pieces. There was no obvious difference on number of peaks between characteristic chromatograms of standard pieces and original pieces, while an obvious difference on number of peaks between chromatograms of control materials and original pieces was found. In addition, standard pieces, original pieces and control materials could generally gather into one class, while raw materials could generally gather into another class. Conclusion: Compared with the raw materials and control materials, the established HPLC characteristic chromatogram of standard pieces can better reflect the internal quality of original pieces, which can be used for the quality control of decoction pieces of Polygoni Multiflori Radix.
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Objective: To optimize the HPLC characteristic spectrum of flavonoid glycosides of Dendrobium officinale,and identify the common and specific components of different provenances. Method: Kromasil 100-5 C18 column was adopted, with tetrahydrofuran-acetonitrile-methanol (10:22:5)-0.05% phosphoric acid as mobile phase (gradient elution). The detection wavelength was 340 nm,the column temperature was 30℃,and the flow rate was 1.0 mL ·min-1. Result: 13 flavonoid characteristic peaks were marked in 27 batches of D. officinale,and 7 characteristic peaks of 6 flavonoid C-glycosides (vicenin Ⅱ,vicenin Ⅰ,schaftoside,isoschaftoside,violanthin and isoviolanthin) and one flavonoid O-glycosides (rutin) was identified. 7-11 characteristic peaks were detected in different batches of samples. Among them,vicenin Ⅱ was a relatively stable common peak in different source samples,and the characteristic peaks of rutin,schaftoside and isoschaftoside were quite different. According to the relative abundance of the characteristic peaks,the samples could be divided into three categories. Among them,the first category had 10 batches of samples,which mainly came from Danxia landforms of Guangdong,Jiangxi,Fujian and Zhejiang (Wuyi) Province (which called "Danxia landform species") and characterized by detection of obvious peak of rutin. The second category had 11 batches of samples,which mainly came from Yunnan and Guangxi Province (which included "Yunnan Guangnan species" and "Guangxi Tiepilan species") and characterized by detection of violanthin and isoviolanthin. And the third category had 6 batches of samples, which were mainly derived from Zhejiang Province (which called "native species from Zhejiang") and characterized by detection of different degrees of rutin peak, but it was difficult to detect the characteristic peaks of violanthin and isoviolanthin. HPLC characteristic chromatograms of D. officinale in bionics wild cultivation and greenhouse of "Danxia landform species" and "Guangxi Tiepilan species" were compared. The results showed that the characteristic peaks in D. officinale planted in greenhouse could be detected stably,which verified the reliability of the source in D. officinale. Conclusion: The analytical method has a better separation effect on flavonoids of D. officinale, with a good reproducibility. The commonness and specificities of flavonoid glycosides components of D. officinale from different categories have basically confirmed. This suggests that Vicenin Ⅱ is suitable to be a reference peak for characteristic chromatogram. Both the relative abundance of rutin and the detection or relative abundance of violanthin and isoviolanthin peaks could be used as a reference to judge the categories of D. officinale in "Danxia landform species" or "Tiepilan species from Yunnan, South Guangdong and Guangxi" or "native species from Zhejiang".