Study on characteristic chromatogram of Chaenomeles sinensis and content determination of 3 flavones / 中国药房

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.

Study on comprehensive quality evaluation of Sargentodoxae Caulis from different habitats / 国际中医中药杂志

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.

Comparison of characteristic chromatogram and the contents of multiple indicator components of Morus alba decoction powder and decoction at different decoction time / 中国药房

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.

Species differences of Baishouwu based on characteristic chromatogram and content determination of acetophenones / 药学实践杂志

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.

Characteristic chromatogram and index components content of substance benchmark of Xuanfu Daizhe Decoction / 中国中药杂志

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.

Stability research of Xiaoer Ganmaoning Oral Liquid based on chromatographic peak matching and image sensory evaluation / 中国中药杂志

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.

Quality standard of Lobeliae Chinensis Herba / 中国中药杂志

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.

Establishment of characteristic chromatogram and content determination of 4 index components in Jianpi yifei biyan prescription standard decoction / 中国药房

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.

HPLC-DAD Characteristic Chromatogram of Baphicacanthis Cusiae Rhizoma et Radix / 中国药学杂志

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.

Comparative study on HPLC characteristic chromatogram of water extracts of Arisaematis Rhizoma and its processed products / 中草药

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.

Comparison of chemical compositions before and after wine-frying of Scutellaria baicalensis based on HPLC characteristic chromatogram, UPLC-Q-TOF/ MS qualitative and multi-component quantitative analysis / 中草药

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.

Study on characteristic chromatogram and determination of indicative components of salt-fried Phellodendri Chinensis Cortex and Yihuang Decoction / 中草药

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.

Identification of Morindae Officinalis Radix root cortex and woody core based on UPLC characteristic chromatogram and chemical pattern recognition method / 中草药

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.

Based on appearance and index components to study on standards of commodity classes of Saposhnikovia Radix / 中草药

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.

HPLC characteristic chromatograms and quantitative analysis of Kunxian Capsules / 中草药

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.

Study on quality evaluation method of classical prescription Mahuang Decoction primary standard substances / 中国中药杂志

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.

Identification based on HPLC and anti-inflammatory targets as well as related constituents analysis of Asarum heterotropoides var. mandshuricum and A. sieboldii / 中国中药杂志

The present work is to establish an HPLC characteristic chromatograms of Asarum heterotropoides var. mandshuricum(AH) and A. sieboldii(AS), combined with cluster analysis for the identification of the two species, and predict their potential anti-inflammatory related targets by network pharmacological method. Eighty-nine samples(12 batches of AS and 77 batches of AH) were analyzed, and 11 characteristic peaks were identified by reference substances, UV spectrum and LC-MS. Cluster analysis showed that AS and AH were divided into two groups, and the ratio of characteristic peak areas can be used to distinguish them. When the ratio of characteristic peak sarisan to kakuol was greater than 5, it was AS, and when the ratio was less than 2, it was AH. The network pharmacological analysis of 119 constituents of Asari Radix et Rhizoma suggested that the anti-inflammatory effect of Asari Radix et Rhizoma might be related to COX-2, COX-1, iNOS, MAPK14, NR3 C1, PPARG and TNF. Among them, COX-2 is a relatively key target, which interacted with the characteristic constituents, asarinin, sesamin, safrole, methyleugenol and sarisan. The characteristic constituents asarinin and sesamin also interacted with the iNOS and MAPK14. Safrole and sarisan can also interact with iNOS, COX-1 and LAT4 H. Methyleugenol also showed interaction with COX-1 and LAT4 H. Since asarinin and sesamin interacted with three targets, COX-2, iNOS and MAPK14, it implied that they were the main active constituents for the anti-inflammatory activity of Asari Radix et Rhizoma. The COX-2 inhibitory activities of asarinin and sesamin were further studied by molecular docking and bioassay. The HPLC method established was simple, feasible and reliable, with predicted anti-inflammatory targets and anti-inflammatory constituents, which could provide a reference for improving the quality evaluation system of Asari Radix et Rhizoma.

Evaluation of key production processes of Schizonepetae Herba formula granules based on characteristic chromatogram and quantitative transfer relationship / 中国中药杂志

The aim of this paper was to evaluate the key production processes of Schizonepetae Herba formula granules based on the new model of combining characteristic chromatogram with quantitative transfer relationship. The rationality of production process design was evaluated by studying the intermediates in different processes of formula granules, analyzing the loss of index component pulegone in each step, and establishing the characteristic chromatogram. The content of pulegone in 10 batches of standard decoction ranged between 0.067% and 0.124%(70%-130% of the average value), and the transfer rate of pulegone was 44.58%-93.97%. After the improvement of the production process, the content of pulegone in Schizonepetae Herba formula granules was 0.093%, and the transfer rate of pulegone was 68.38%, which was consistent with the parameters range of standard decoction. This study emphasized the integrality of the research process of traditional Chinese medicine(TCM) formula granules, and provided a new idea for the quality control of TCM with content determination as the main evaluation index for a long time.

Evaluation of key production processes of Chan Taoren formula granules based on characteristic chromatogram and D-amygdalin transfer rate / 中国中药杂志

This study aimed to establish characteristic chromatogram and content determination method for Chan Taoren formula granules,evaluate the production processes of Chan Taoren formula granules based on the correlation of characteristic chromatogram and the transfer rate of D-amygdalin,and clarify the key control points. The optimized analytical method was carried out on a Waters CORTECS C18 column(4. 6 mm×150 mm,2. 7 μm) with acetonitrile-0. 1% phosphoric acid aqueous solution as the mobile phase at a flow rate of 0. 6 m L·min-1. The detection wavelength was 207 nm,and the column temperature was 20 ℃ ． As compared with the standard decoction of Chan Taoren,there were five characteristic peaks in the decoction pieces,extracts,concentrates,spray-dried powders and formula granules,basically consistent in relative retention time and peak pattern; in addition,the transfer rate of D-amygdalin from Chan Taoren pieces to the formula granules was within the transfer rate range of standard decoction. The average transfer rate of D-amygdalin was 56.65%,72.85%,94.58% and 99.29% respectively in the extraction,concentration,spray drying and granulation processes. Therefore,the factors affecting D-amygdalin in the extraction process were further studied. The results showed that D-amygdalin was easily converted to L-amygdalin in the water extraction process,leading to a low transfer rate of D-amygdalin in this process.D-amygdalin was unstable under alkaline conditions and prone to isomerization. Both liquid to solid ratio and extraction time had significant effects on the extraction rate of D-amygdalin. In this study,the key links in the production process of Chan Taoren formula granules was clarified based on the characteristic chromatogram and the quantity transmission of D-amygdalin,which provided a theoretical basis for production and quality control.

Optimization of Purification Process of Total Alkaloid Extract of Berberis dictyophylla Cortex by Macroporous Resin and Investigation of Its Quality Standard / 中国实验方剂学杂志

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.