Fingerprint establishment of Huangqin decoction and study on spectrum-effect relationship of its antidermatophytic activity in different phase states / 中国药房

OBJECTIVE To establish the fingerprint of Huangqin decoction （HQD）， to separate the phase states and screen the active phase states of antidermatophytic activity so as to study the spectrum-effect relationship. METHODS HPLC method was adopted using baicalin as reference， the fingerprints of 10 batches of HQD were drawn and the similarity evaluation was carried out using the Similarity Evaluation System of Chromatographic Fingerprint of TCM （2012 edition） to determine the common peak； the phase states of HQD were separated and characterized by high-speed centrifugation and membrane dialysis. The minimum inhibitory concentrations （MIC） of HQD and its different phase states against Trichophyton mentagrophytes were determined simultaneously. Using the peak area of 37 common peaks as independent variable， MIC as dependent variable， Pearson correlation analysis was performed by using SPSS 21.0 software. RESULTS A total of 37 common peaks were obtained in HPLC fingerprints of 10 batches of HQD， with the similarity higher than 0.99. Ten components were identified， such as albiflorin， paeoniflorin， liquiritin apioside， baicalin， melaleuca glycoside A， wogonoside， baicalein， glycyrrhizic acid， wogonin and oroxylin A. HQD was split into 3 phase states， such as precipitation phase （HQD-P）， solution phase （HQD-S） and nano phase （HQD-N）. The morphology of HQD-P was irregular granular， and the average particle size was 4.670-91.522 μm. The morphology of HQD-S was uniform flakes， and no particle size was detected. HQD-N was spherical in shape and the particle size was （129.0±12.9） nm. MIC values of each phase state of HQD against T. mentagrophytes in different phase states were HQD-N （4.64 mg/mL） ＜HQD （5.85 mg/mL） ＜HQD-P （7.37 mg/mL） ＜HQD-S （12.89 mg/mL） at the same dosage. Pearson correlation analysis showed that the peak area of 25 of the 37 common peaks （including identified components） was significantly negatively correlated with MIC （absolute values of correlation coefficient＞0.95 and P＜0.05）. CONCLUSIONS The chemical composition of 10 batches of HQD is consistent； HQD-N is the active phase state of HQD. Ten components such as paeoniflorin， liquiritin apioside and baicalin may be the main active components of HQD. The antidermatophytic effect of HQD is closely related to its component content and physical phase state.

Screening of active ingredients for estrogenic effect of total flavonoids from Cuscutae Semen / 中国药房

OBJECTIVE To screen the active ingredient with estrogenic effect from total flavonoids of Cuscutae Semen. METHODS The estrogenic effect of total flavonoids from 10 batches of Cuscutae Semen was evaluated with mouse uterus coefficient and endometrial thickness as evaluation indexes， establish its fingerprint and calibrate the common peak. Common peak and spectrum-effect relationship of the above two indicators were analyzed by bivariate relationship analysis and grey correlation analysis to screen active components with estrogenic effect. UPLC-Q-TOF-MS technology was used to characterize the active components. RESULTS The estrogenic effect of total flavonoids from 10 batches of Cuscutae Semen was good. Twenty-eight and thirty-three common peaks of total flavonoids in Cuscutae Semen were obtained in the positive and negative ion modes respectively. The constituents represented by peaks 7，10，12-16，26 in positive ion mode and peaks 2，5，8，9，12，16，19，22-26 in negative ion mode were highly correlated with the estrogenic effect of total flavonoids from Cuscutae Semen. Further identification showed that the active substances with estrogenic effect from the total flavonoids of Cuscutae Semen were 5，7，3′， 4′-tetramethoxyflavone， 6- O-（trans） p-coumarin-furanfructose-（2→1）-glucopyranoside， rutin， kaempferol-3，7-diglucoside， apigenin-7-O-glucoside， hyperoside， baicalin， quercitin， quercetin， apigenin， kaempferol， isorhamnetin， rhododendron， isoquercetin， kaempferol-3-furan arabinoside， 2，6-octadecanediacetylic acid. CONCLUSIONS A total of 16 chemical components with estrogenic effect are screened from total flavonoids of Cuscutae Semen in the study， which can provide reference for the development of phytoestrogens.

Application of partial least squares algorithm to explore bioactive components of crude and stir-baked hawthorn for invigorating spleen and promoting digestion / 中国中药杂志

This study was aimed at identifying the bioactive components of the crude and stir-baked hawthorn for invigorating spleen and promoting digestion, respectively, to clarify the processing mechanism of hawthorn by applying the partial least squares(PLS) algorithm to build the spectrum-effect relationship model. Firstly, different polar fractions of crude and stir-baked hawthorn aqueous extracts and combinations of different fractions were prepared, respectively. Then, the contents of 24 chemical components were determined by ultra-high performance liquid chromatography-mass spectrometry. The effects of different polar fractions of crude hawthorn and stir-baked hawthorn aqueous extracts and combinations of different fractions were evaluated by measuring the gastric emptying rate and small intestinal propulsion rate. Finally, the PLS algorithm was used to establish the spectrum-effect relationship model. The results showed that there were significant differences in the contents of 24 chemical components for different polar fractions of crude and stir-baked hawthorn aqueous extracts and combinations of different fractions, and the gastric emptying rate and small intestinal propulsion rate of model rats were improved by administration of different polar fractions of crude and stir-baked hawthorn aqueous extracts and combinations of different fractions. The bioactive components of crude hawthorn identified by PLS models were vitexin-4″-O-glucoside, vitexin-2″-O-rhamnoside, neochlorogenic acid, rutin, gallic acid, vanillic acid, citric acid, malic acid, quinic acid and fumaric acid, while neochlorogenic acid, cryptochlorogenic acid, rutin, gallic acid, vanillic acid, citric acid, quinic acid and fumaric acid were the bioactive components of stir-baked hawthorn. This study provided data support and scientific basis for identifying the bioactive components of crude and stir-baked hawthorn, and clarifying the processing mechanism of hawthorn.

Optimization of processing methods for Terminalia chebula based on fingerprint，antioxidant spectrum-effect correlation and multi-component contents / 中国药房

OBJECTIVE To compare the similarities and differences between raw and different preparations of Terminalia chebula based on fingerprint， antioxidant spectrum-effect correlation and multi-component contents， and to provide a reference for searching for modern processing methods of T. chebula that are similar to classical ancient methods. METHODS Ten batches of raw and different preparations of T. chebula （single stir-fried products， bran-roasted products， sand-scorched products， ash-roasted products， stir-fried charcoal products， and wine-steamed products） were used as test samples. The high-performance liquid chromatography （HPLC） fingerprints of different samples were established by using the Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 edition）， the chromatographic peaks were identified， and chemometrics analysis was carried out. At the same time， HPLC method was used to determine the contents of 8 identified components. The antioxidant capacity of raw and different preparations of T. chebula was determined by DPPH free radical scavenging method， and the spectrum- effect relationship was analyzed. RESULTS A total of 20 common peaks were identified in the fingerprints of the raw and different preparations of T. chebula， and the similarity of each sample was ＞0.9. Nine common peaks were identified from the raw and different preparations of T. chebula， including chromatographic peak 2 （chebulic acid）， 3 （gallic acid）， 6 （punicalagin A）， 8 （punicalagin B）， 12 （corilagin）， 15 （chebulagic acid）， 18 （ellagic acid）， 19 （1，2，3，4，6-O-pentagalloyl glucose）， 20 （chebulinic acid）， etc. Compared with crude drug， the contents of the above 8 components （punicalagin A and B are recorded as punicalagin） in different preparations of T. chebula were changed， and the changes of the contents of the stir-fried charcoal and wine-steamed products were more obvious than those of other processed products. Chemometric analysis showed that the fingerprints of stir-fried charcoal and wine-steamed products of T. chebula were obviously distinguished from other processed products， and the fingerprint information of raw products and other processed products of T. chebula was partially overlapped. Four main differential components （chebulinic acid， chebulagic acid， gallic acid， ellagic acid） were obtained between raw and processed products of T. chebula； and four main effective components （chebulinic acid， chebulagic acid， gallic acid， corilagin） were obtained by analyzing the spectrum-effect relationship of antioxidant activity. The single stir-fried product of T. chebula showed the strongest antioxidant activity. CONCLUSIONS The single stir-frying method is a modern processing method of T. chebula which is similar to the classical ancient method and is more excellent.

Screening the effective components in treating dampness stagnancy due to spleen deficiency syndrome and elucidating the potential mechanism of Poria water extract / 中国天然药物

Poria is an important medicine for inducing diuresis to drain dampness from the middle energizer. However, the specific effective components and the potential mechanism of Poria remain largely unknown. To identify the effective components and the mechanism of Poria water extract (PWE) to treat dampness stagnancy due to spleen deficiency syndrome (DSSD), a rat model of DSSD was established through weight-loaded forced swimming, intragastric ice-water stimulation, humid living environment, and alternate-day fasting for 21 days. After 14 days of treatment with PWE, the results indicated that PWE increased fecal moisture percentage, urine output, D-xylose level and weight; amylase, albumin, and total protein levels; and the swimming time of rats with DSSD to different extents. Eleven highly related components were screened out using the spectrum-effect relationship and LC-MS. Mechanistic studies revealed that PWE significantly increased the expression of serum motilin (MTL), gastrin (GAS), ADCY5/6, p-PKAα/β/γ cat, and phosphorylated cAMP-response element binding protein in the stomach, and AQP3 expression in the colon. Moreover, it decreased the levels of serum ADH, the expression of AQP3 and AQP4 in the stomach, AQP1 and AQP3 in the duodenum, and AQP4 in the colon. PWE induced diuresis to drain dampness in rats with DSSD. Eleven main effective components were identified in PWE. They exerted therapeutic effect by regulating the AC-cAMP-AQP signaling pathway in the stomach, MTL and GAS levels in the serum, AQP1 and AQP3 expression in the duodenum, and AQP3 and AQP4 expression in the colon.

Analysis of the spectrum-effect relationship of different processed products of Epimedii folium on cytotoxicity / 国际中医中药杂志

Objective:To explore the effects of different processed products of Epimedii Folium on cytotoxicity and the material basis of toxicity. Methods:By using the SRB method to investigate the effects of different processed products of Epimedii Folium on the proliferation of HaCaT cells; and based on the grey correlation analysis method to establish the data spectrum effect relationship of HPLC fingerprint spectrum-toxicity so as to determine the toxic components and processing methods. Results:The value of cytotoxicity IC 50 of different processed products of Epimedii Folium is as follow: vinegar fried> oil fried > original > single fried > salt fried > wine fried. Among the 12 characteristic chromatographic peaks, Peak No.3 (magnoflorine, correlation value: 0.870) and Peak No.6 (epimedin C, correlation value: 0.851) are highly correlated with the toxicity value. Conclusions:Both vinegar fried and oil fried Epimedii Folium have the effect of reducing toxicity. Magnoflorine and epimedin C may be the main toxic components in Epimedii Folium. The study provides scientific basis for the research on the process optimization of Epimedii Folium concocting to reduce toxicity.

Spectrum-effect relationship of anti-gastric ulcer effect of Shaoyao gancao decoction / 中国药房

OBJECTIVE To study the sp ectrum-effect relationship of anti-gastric ulcer effect of Shaoyao gancao decoction. METHODS Eleven batches of Shaoyao gancao decoction were prepared ；gastric ulcer model was established by anhydrous ethanol modeling method. Using Weikangling as positive control ，the effects of Shaoyao gancao decoction on the contents of defensive factors [nitric oxide （NO），epidermal growth factor （EGF），superoxide dismutase （SOD）] and attack factor [malondialdehyde （MDA）] in gastric tissue of model rats were investigated. HPLC fingerprints of 11 batches of Shaoyao gancao decoction were established and similarity evaluation was performed with Similarity Evaluation System of Traditional Chinese Medicine Chromatographic Fingerprint （2004A edition ）； common peaks were identified by comparing with mixed control. The spectrum-effect relationship of Shaoyao gancao decoction against gastric ulcer was analyzed based on the grey correlation analysis. RESULTS Eleven batches of Shaoyao ganyao decoction could significantly decrease the content of MDA in gastric tissue ，while increased the contents of NO ，EGF and SOD in gastric ulcer model rats （P＜0.01），and had a certain inhibitory effect on the gastric ulcer. There were 23 common peaks in chromatograms of 11 batches of samples ，and the similarity with the control fingerprint was not less than 0.9. By comparing with mixed control ，7 common peaks were identified ，namely gallic acid （peak 5），albiflorin（peak 9），paeoniflorin（peak 10），liquiritin（peak 12），isoliquiritin apioside （peak 14），isoliquiritoside（peak 15）， glycyrrhizic acid （peak 22）. The average correlation degree of 7 identified common peaks and pharmacodynamic indexes were greater than 0.6，of which peak 22（glycyrrhizic acid ），peak 10（paeoniflorin）and peak 12（liquiritin）had the largest correlation ， and their values were 0.807，0.772 and 0.770 respectively. RESULTS The anti-gastric ulcer effect of Shaoyao gancao decoction is the result of the synergistic effect of multiple components ，among which glycyrrhizic acid ，paeoniflorin and liquiritin may be the main pharmacodynamic components.

Study on anticoagulant material basis and mechanism of Trichosanthis Semen and its shell and kernel based on spectrum-effect relationship integrated molecular docking / 中国中药杂志

This study explored the anticoagulant material basis and mechanism of Trichosanthis Semen and its shell and kernel based on spectrum-effect relationship-integrated molecular docking. High performance liquid chromatography(HPLC) fingerprints of Trichosanthis Semen and its shell and kernel were established. Prothrombin time(PT) and activated partial thromboplastin time(APTT) in mice in the low-and high-dose(5, 30 g·kg~(-1), respectively) Trichosanthis Semen, the shell, and kernel groups were determined as the coagulation markers. The spectrum-effect relationship and anticoagulant material basis of Trichosanthis Semen and its shell and kernel were analyzed with mean value calculation method of Deng's correlation degree(MATLAB) and the common effective component cluster was obtained. Then the common targets of the component cluster and coagulation were retrieved from TCMSP, Swiss-TargetPrediction, GenCLiP3, GeneCards, and DAVID, followed by Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment of the targets. The main anticoagulant molecular mechanism of the component cluster was verified by SYBYL-X 2.1.1. The spectrum-effect relationship of Trichosanthis Semen and its shell and kernel was in positive correlation with the dosage. The contribution of each component to anticoagulation was not the same, suggesting that the material basis for anticoagulation was different, but they have common effective components(i.e. common material basis), such as adenine(peak 3), uracil(peak 4), hypoxanthine(peak 6), xanthine(peak 9), and adenosine(peak 11). Network pharmacology showed that these components can act on multiple target proteins such as NOS3, KDR, and PTGS2, and exert anticoagulant effect through multiple pathways such as VEGF signaling pathway. They involved the biological functions such as proteolysis, cell component such as cytosol, and molecular functions. The results of molecular docking showed that the binding free energy of these components with NOS3(PDB ID: 1 D0 C), KDR(PDB ID: 5 AMN), and PTGS2(PDB ID: 4 COX) was ≤-5 kJ·mol~(-1), and the docking conformations were stable. Spectrum-effect relationship-integrated molecular docking can be used for the optimization, virtual screening, and verification of complex chemical and biological information of Chinese medicine. Trichosanthis Semen and its shell and kernel have the common material basis for anticoagulation and they exert the anticoagulant through multiple targets and pathways.

Contributions of flavonoids from citri reticulatae pericarpium to gastric hormones, CD3~+ and TFF3 mRNA expression in rats with spleen deficiency intervened by Liujunzi Decoction / 中国中药杂志

The present study established the spectrum-effect relationship model of flavonoids in Citri Reticulatae Pericarpium(CRP) from 15 batches of Liujunzi Decoction and statistically analyzed the correlation between chemical peaks and efficacy to identify the main effective components. HPLC fingerprints of flavonoids in CRP from 15 batches of Liujunzi Decoction were established. HPLC analysis was carried out on the Venusil XBP C_(18)(L) column(4.6 mm×250 mm, 5 μm) at 30 ℃ with acetonitrile-water(containing 0.1% formic acid) as mobile phase for gradient elution, a flow rate of 1.0 mL·min~(-1), and detection wavelength of 300 nm to obtain chemical fingerprints. Additionally, the effects of flavonoids from CRP in 15 batches of Liujunzi Decoction on the content of GAS, MTL, and VIP, TFF3 mRNA expression, and percentage of CD3~+ T-cells of model rats with spleen deficiency were determined. The spectrum-effect relationship model was established by gray correlation analysis. The results showed that the main characteristic peaks with great contribution to the regulation of gastrointestinal tract were peak 16(vicenin-2), peak 63(sinensetin), peak 64(isosinensetin), peak 65(nobiletin), peak 67(3,5,6,7,8,3',4'-heptemthoxyflavone), peak 68(tangeretin), and peak 69(5-desmethylnobiletin). Therefore, there was a linear correlation between flavonoids from CRP in Liujunzi Decoction and the efficacy, and the medicinal effect was achieved by multi-component action. This study is expected to provide a new idea for exploring the material basis of the effect, i.e., regulating qi prior to replenishing qi, of CRP in Liujunzi Decoction.

Establishment of carotenoid fingerprint in Lycium barbarum and study on its antioxidant activity spectrum- effect relationship / 中国药房

OBJECTI VE To establish the high performan ce liquid c hromatography（HPLC）fingerprint of carotenoid in Lycium barbarum，and to investigate the spectrum-effect relationship between its common peak and antioxidant activity. METHODS HPLC method was adopted. The fingerprints of carotenoid in 34 batches of L. barbarum from different producing areas were established by Similarity Evaluation System of TCM Fingerprint （2012 edition），and similarity evaluation and common peak identification were carried out. Taking scavenging rate of DPPH free radical as index ，in vitro antioxidant activity of carotenoid in L. barbarum was investigated. The spectrum-effect relationship between the common peaks of carotenoids in L. barbarum and antioxidant activity was analyzed by grey correlation method. RESULTS There were 4 common peaks in the fingerprints of carotenoids in 34 batches of L. barbarum ，and the similarity was not less than 0.903. Peak 1 was identified as zeaxanthin ，and peak 4 as zeaxanthin dipalmitate. The scavenging rates of them to DPPH free radical were 1.792％-3.160％. The common peaks of carotenoids in L. barbarum were positively correlated with scavenging rate of DPPH free radical ，and the correlation degree was greater than 0.6；the correlation degree of peak 2 and peak 4（zeaxanthin dipalmitate ）with scavenging rate of DPPH free radical was greater than 0.8. According to the correlation degree ，the contribution of each common peak to scavenging rate of DPPH free radical was determined as peak 2＞ peak 4（zeaxanthin dipalmitate ）＞peak 1（zeaxanthin）＞peak 3. CONCLUSIONS In this study ，HPLC fingerprint of carotenoid in L. barbarum is successfully established ，and two common peaks are identified. The chemical components represented by peak 2 and zeaxanthin palmitate may be the material basis of antioxidant activity of carotenoid in L. barbarum .

Screening of effective antioxidant components from Trichosanthes extract and assessment of their antioxidant activity / 南方医科大学学报

OBJECTIVE@#To screen the effective antioxidant components in Trichosanthes extract based on the mean value of Deng's correlation degree and assess the antioxidant activity of the identified components.@*METHOD@#High-performance liquid chromatography (HPLC) was used to obtain the fingerprints of Trichosanthes extract, and the clearance rates of DPPH · and O2-· by 3, 9 and 27 mg/mL Trichosanthes extract were determined. The antioxidant spectrum effect of Trichosanthes extract was analyzed by calculating the mean value of Deng's correlation degree to screen the effective antioxidant component group. According to the contents of each known components in the antioxidant effective component group, mixed solutions of the components were prepared and tested for their clearance rates of DPPH · and O2-·.@*RESULTS@#The 36 common peaks in HPLC fingerprints of Trichosanthes extract showed different degrees of correlation with DPPH · and O2-· clearance. The common peaks with a correlation degree greater than the median value included peaks 21, 36, 8, 31, 14, 5, 27, 2, 24, 15, 18, 33, 22, 34, 35, 19, 28 and 25. The 5 components, namely kaempferol (peak 36), isoquercitrin (peak 8), luteolin (peak 31), rutin (peak 5) and apigenin (peak 35), were tentatively identified to constitute the effective antioxidant component group with a mass ratio 3∶2∶2∶ 1∶1 in Trichosanthes extract. The prepared mixed solutions of antioxidant effective component group (6.12, 2.04, and 0.68 μg/mL) showed clearance rates of DPPH · of 65.4%, 64.0% and 61.0%, and clearance rates of O2-· of 12.9%, 9.5% and 8.3%, respectively.@*CONCLUSION@#We identified the material basis for the antioxidant activity of Trichosanthes and screened the antioxidant effective component group in Trichosanthes extract.

Establishment of HPLC Fingerprint of Different Polar Parts of Zhuang Medicine Calonyction muricatum and Spectrum-effect Relationship of Its Analgesic and Anti-inflammatory Effects / 中国药房

OBJECTIVE：To establish HPLC fingerprints of different polar parts of Zhuang medicine Calonyction muricatum ， and to study its spectrum-effect relationship with analgesic and anti-inflammatory effects. METHODS ：The total part ，ethyl acetate part，n-butanol part and water part of C. muricatum were prepared. HPLC fingerprints of different polar parts were established by HPLC method combined with the Similarity Evaluation System of TCM Chromatogramtic Fingerprint （2012A），and the common peaks were identified. Using writhing times and ear swelling degree in mice as analgesic and anti-inflammatory indexes ，analgesic and anti-inflammatory activity of different polar parts of C. muricatum were investigated. The correlation of the common peaks of HPLC fingerprint with analgesic and anti-inflammatory indexes was analyzed by grey correlation analysis ，bivariate correlation analysis and partial least square （PLS） method. RESULTS ： There were 11 common peaks for the different polar parts of C. muricatum ，and 5 components were identified by reference comparison，i.e. neochlorogenic acid （peak 3），chlorogenic acid （peak 5）， cryptochlorogenic acid （peak 6）， isochlorogenic acid A （peak 10），isochlorogenic acid C （peak 11）. The grey correlation analysis showed that the correlation between all common peaks and analgesic and anti- inflammatory effects were greater than 0.6 （except the correlation between peak 6 and analgesic effects ），showing correlation relationship ；the correlation of peaks 3，7 and 10 with analgesic and anti-inflammatory effects were all greater than 0.8，which was highly related. Bivariate correlation analysis showed that the correlation of peak 1，3，4，7，9，10，11 with analgesic and anti-inflammatory effects were all greater than 0.6，showing correlation relationship. PLS method showed that peaks 1，3，4，7，9，10，11 contributed greatly to playing an analgesic and anti-inflammatory role. CONCLUSIONS ：HPLC fingerprints of different polar parts of C. muricatum is established and five common peak components were identified. Neochlorogenic acid ，isochlorogenic acid A ，isochlorogenic acid C and chemical components represented by peaks 1，4，7，9 may be the pharmacodynamic substances of C. muricatum to exert analgesic and anti-inflammatory effects.

Establishment of UPLC Fingerprint of Poria cocos Aqueous Extract and Study on Its Spectrum-effect Relationship with Sedative and Hypnotic Effect / 中国药房

OBJECTIVE：To establis h the UPLC fingerprint of Poria co cos aqueous extract ，and to investigate its relationship with sedative and hypnotic effect. METHODS ：Ten batches of P. cocos from different areas were extracted with water to obtain the aqueous extract. UPLC method was adopted. The determination was performed on Waters HSS-C 18 column with mobile phase consisted of 0.1％ phosphoric acid solution-acetonitrile-methanol （gradient elution ） at the flow rate of 0.4-0.2 mL/min. The detection wavelengths were set at 210 and 242 nm. The column temperature was 40 ℃，and sample size was 2 μL. The fingerprints of 10 batches of P. cocos aqueous extracts were established by using the Similarity Evaluation System of TCM Fingerprint （2012A version），and the common peaks were identified. The sedative and hypnotic effects of 10 batches of P. cocos aqueous extracts from different areas under the synergistic action of pentobarbital sodium were investigated by taking the sleeping rate ，sleep latency and sleep duration of mice as the single efficacy index. After data transformation of single efficacy index and total efficacy （single indexes calculated by analytic hierarchy process ），grey correlation analysis was used to analyze the correlation between the common peaks in fingerprint of P. cocos aqueous extract and the single efficacy index and total efficacy. RESULTS ：There were 24 common peaks in 10 batches of aqueous extract of P. cocos ， and 11 components were identified ， i.e. 16 α-hydroxydehydrotrametenolic acid （peak 6），16α-hydroxytrametendic acid （peak 7），poricoic acid B （peak 9），dehydrotumulosic acid（peak 10），poricoic acid A （peak 12），polyporenic acid C （peak 15），3-O-acetyl-16α-hydroxydehydrotrametenolic acid （peak 17），dehydropachymic acid （peak 20），pachymic acid （peak 21），dehydrotrametenolic acid （peak 22），dehydroeburicoic acid （peak 24）. Grey correlation analysis showed ，the correlation between 24 peaks and sleep duration was greater than 0.6（0.611 5- 0.811 8）；the correlation between 24 peaks and sleep latency was greater than 0.6（0.605 9-0.790 4），except for peaks 14，24 and 2；the correlation of 24 peaks between sleeping rate was greater than 0.6（0.606 4-0.721 6），except for peaks 23，19，17 and 5； the correlation of 24 peaks between total efficacy was greater than 0.6（0.619 0-0.781 2），except for peaks 2，5，19. The top 10 chromatographic peaks related to the total efficacy were peak 15（polyporenic acid C ），peak 16，peak 8，peak 11，peak 12 （poricoic acid A ）， peak 1， peak 7 （16 α-hydroxytrametendicacid）， peak 3， peak 9 （poricoic acid B ） and peak 20 （dehydropachymic acid ）. CONCLUSIONS ：UPLC fingerprint of P. cocos aqueous extract was established and 11 components were identified. Ten components such as polyporus acid C are closely related to the total efficacy of sedation and hypnosis ，which preliminarily reveal the material basis of the sedative and hypnotic effect of P. cocos .

Material Basis and Mechanism of Epimedii Folium Against Osteoporosis Based on Spectrum-Effect Relationship and Network Pharmacology / 中国实验方剂学杂志

Objective:To study the material basis and potential molecular mechanism of Epimedii Folium against osteoporosis. Method:The chemical components in 14 batches of Epimedii Folium were analyzed by ultra-performance liquid chromatography-quadrupole-time of flight-tandem mass spectrometry （UPLC-Q-TOF-MS/MS）. With the activity of alkaline phosphatase （ALP） as the pharmacodynamic index，the partial least squares regression analysis （PLSR） was conducted to establish a model uncovering the spectrum-effect relationship between UPLC-Q-TOF-MS/MS spectral peak and ALP activity and screen the active components against osteoporosis. Online databases such as the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP），Comparative Toxicogenomics Database （CTD），Database for Annotation，Visualization，and Integrated Discovery （DAVID） and Cytoscape 3.6.1 were employed to predict the possible mechanism of action of Epimedii Folium against osteoporosis. Result:A total of 61 peaks and 56 compounds were identified by UPLC-Q-TOF-MS/MS. PLSR showed that icariin，baohuoside Ⅰ，epimedin A，sagittatoside A，and baohuoside Ⅱ might be the active components for Epimedii Folium to inhibit osteoporosis considering their strong correlation with ALP activity. As revealed by the network pharmacological analysis of the five components mentioned above，Epimedii Folium<italic> </italic>mainly regulated seven targets such as tumor necrosis factor （TNF），androgen receptor （AR），and estrogen receptor 1 （ESR1） and eight key pathways like endocrine and other factor-regulated calcium reabsorption，vascular endothelial growth factor （VEGF） signaling pathway，and transient receptor potential （TRP） channels to exert its anti-osteoporosis effect. Conclusion:The exploration of material basis and potential molecular mechanism of Epimedii Folium against osteoporosis based on UPLC-Q-TOF-MS/MS，spectrum-effect relationship，and network pharmacology has provided an experimental basis for the scientific explanation and clinical application of Epimedii Folium in treating osteoporosis.

Study on Spectrum-effect Relationship of Anti-inflammatory and Analgesic Effects of Ethyl Acetate Extract from Zhuang Medicine Stahlianthus involucratus / 中国药房

OBJECTIVE：To i nvestigate the spectrum-effect relationship of analgesic and anti-inflammatory effects of ethyl acetate extract from Zhuang medicine Stahlianthus involucratus from different habitats. METHODS ：Ten batches of S. involucratus from different habitats were used as samples to investigate the anti-inflammatory and analgesic activities of ethyl acetate extracts by xylene induced ear swelling test and acetic acid induced writhing test in mice. HPLC fingerprints of 10 batches of ethyl acetate extract from S. involucratus were established and their similarity was evaluated by using Similarity Evaluation System of TCM Chromatogram Fingerprint （2012 edition），and the common peaks were identified by comparison with the control. The spectrum-effect relationship of anti-inflammatory and analgesic effects of ethyl acetate extract from S. involucratus were analyzed on the basis of Pearson correlation coefficient （auricle swelling degree and writhing times in 15 min as pharmacodynamic indexes ）and Grey relational analysis （inhibition rate of ear swelling and analgesic rate as pharmacodynamic indexes ）. RESULTS ： batches of ethyl acetate extract from S. involucratus had obvious anti-inflammatory and analgesic effects ； inhibition rates of ear swelling in mice were 46.43％-55.16％，and the analgesic rates of mice were 45.56％-52.72％. A total of 18 common peaks were identified in 10 batches of samples ，andthe similarity between them and the control fingerprint was 0.994-0.997. Compared with substance control ，the pea ks 1，2 and 4 were identified as protocatechuic acid ， p-hydroxy- 0771-4953513。E-mail：liangjie1101@126.com benzoic acid and p-hydroxybenzaldehyde，respectively. Results of Pearson correlation analysis showed that peak 10 and peak 18 were significantly negative correlated with auricle swelling degree and writhing times in 15 min（r were values －0.853，－0.738，P values were 0.002，0.015，respectively）. Results of Gray correlation degree analysis showed that the correlation degree of 18 common peaks with inhibition rate of ear swelling and analgesic rate were all greater than 0.65；among them ，peaks 14，1（protocatechuic acid ），17，9，4（p-hydroxybenzaldehyde），2（p-hydroxybenzoic acid ）， 16，7 and 6 showed the relatively high correlation degree （correlation degree ＞0.7）；peak 1（protocatechuic acid ），17，14，9，16，2 （p-hydroxybenzoic acid ）and 4（p-hydroxybenzaldehyde）showed the relatively high correlation degree （correlation degree ＞0.7）. CONCLUSIONS：The ethyl acetate extract of S. involucratus show good anti-inflammatory and analgesic effects. Peak 1 （protocatechuic acid ），2（p-hydroxybenzoic acid ），10，14，17，18 may be its main active ingredients.

Stoichiometric Analysis in Fingerprint of Aqueous Extract of Citri Reticulatae Pericarpium and Spectrum-effect Relationship with Diabetic Cognitive Dysfunction / 中国实验方剂学杂志

<b>Objective::To establish the HPLC fingerprint of aqueous extract of Citri Reticulatae Pericarpium, and research the spectrum-effect relationship in diabetic cognitive dysfunction rats. <b>Method::The HPLC fingerprints of 21 batches of Citri Reticulatae Pericarpium were established, with acetonitrile (A)-0.1%of phosphoric acid (B) as the flow phase for gradient elution. The detection wavelength was 270 nm. Acomparative analysis was performed for the results based on similarity evaluation, SPSS 24.0 two-dimension clustering analysis and SIMCA 14.1 principal component analysis of fingerprint software. Thediabetic rat model wasduplicated by one-time intraperitoneal injection with streptozotocin (STZ) at the dose of 50 mg·kg^-1. After successful replication of the model, thediabetic rats were given aqueous extract of Citri Reticulatae Pericarpium by gavage. The blood glucose value was determined at regular intervals. The cognitive function was analyzed with Morris water maze experiment at the 12 week. Grey relational analysis (GRA) and partial least squares regression analysis were used to study the spectrum-effect relationship. <b>Result::Totally 16 common peaks of water extract of Citri Reticulatae Pericarpium were confirmed, five substances were identified, and 21 species of Citri Reticulatae Pericarpium of different varieties and origins were divided into 2 categories. Compared with the model group, the blood glucose of rats in each group treated with water extract of Citri Reticulatae Pericarpiumwere reduced, and their cognitive function was improved.According to the grey correlation analysis and the least partial square regression analysis, the peaks No. 19, 15, 4, 17 and 6 were significantly correlated with the cognitive function of diabetic rats. <b>Conclusion::There were some differences in aqueous extract of Citri Reticulatae Pericarpium between different varieties and origins. It is convenient and fast to make a comprehensive quality evaluation of aqueous extract of Citri Reticulatae Pericarpium of different varieties and producing areas by stoichiometry, and aqueous extract of Citri Reticulatae Pericarpium can reduce the blood glucose level and improve the cognitive function of diabetic cognitive dysfunction rats, its mechanisms of action are resultedfrom the joint effects of multiple effective components, and peaks No. 19, 15, 4, 17 and 6 are components closely related to the cognitive function of diabetic rats.

Establishment of Fingerprint and Spectrum-effect Relationship Study on Anti-oxidantion Activity of Anemarrhena asphodeloides and Its Different Processed Products / 中国药房

OBJECTIVE：To establish the fingerprint of ethanol extract and acetone extract from Anemarrhena asphodeloides and its different processed products ，and to investigate the spectrum-effect relationship between the fingerprint and the antioxidant activity. METHODS ：HPLC method and HPLC-ELSD method were adopted. The determination was performed on Thermo BDS Hypersil C 18 column with mobile phase consisted of acetonitrile- 0.2％ acetic acid at the flow rate of 1.0 mL/min. The column temperature was 30 ℃，and the detection wavelength was set at 258 nm. The sample size was 10 μL. The determination was performed on XDB-C 18 columnwith mobile phase consisted of acetonitrile-0.1％ acetic acid （gradient elution ）at the flow rate of 0.9 mL/min. The column temperature was 30 ℃ . The temperature of atomizer was 40 ℃ and the flow rare of N 2 was 1.6 mL/min. The sample size was 10 μL. Using mangiferin and timosaponin B Ⅱ as reference ，Fingerprint Similarity Eva- com luation System of TCM Chromatogram （2004A edition ）was adopted to draw the fingerprint of ethanol extract and acetoneextract from 20 batches of A. asphodeloides and its different processed products to confirm common peaks. Using scave nging rate of 1，1-diphenyl-2-trinitrophenylhydrazine（DPPH）radical as index，antioxidant activities of ethanol extract and acetone extract from 20 batches of A. asphodeloides and its processed products were investigated. Using scavenging rate of DPPH radical as dependent variable ，common peak area as independent variable ，PLSR was used to analyze the spectrum-effect relationship of ethanol extract and acetone extract from A. asphodeloides with antioxidantion activity. RESULTS ：Eight peaks （M1-M8）were identified in the fingerprints of ethanol extracts from 20 batches of processed A. asphodeloides . Mangiferin （chromatogram peak M 7）was identified with similarity of 0.389-1.000；seven comon peaks （S1-S7）and timosaponin B Ⅱ（peak S 5）were identified in the fingerprint of acetone extract ，and the similarity was 0.044-0.999. DPPH radical scavenging rate of ethanol extract from 20 batches of A. asphodeloides and its processed products was 21.23％- 81.39％，and A. asphodeloides was significantly lower than salt-processed A. asphodeloides with salt wine-processed A. asphodeloides （P＜0.001）；and that of acetone extract was 49.73％-83.78％，and A. asphodeloides was significantly higher than stir-baked A. asphodeloides with salt ，wine or fire （P＜0.001）. The standardized regression coefficients of peaks M 2-M7 in the spectrum of ethanol extract from A. asphodeloides were all greater than 0，which was positively correlated with antioxidant activity. Only the variable importance projection （VIP）value of peak M 7 was greater than 1，which had an important contribution. The standardized regression coefficients of peaks S 4-S7 in the acetone extract spectrum of A. asphodeloides were greater than 0，and were positively correlated with antioxidant activity. The order of VIP values was peak S 5＞S6＞S4，and the VIP values were all greater than 1. CONCLUSIONS：The fingerprint of the different processed products A. asphodeloides and its antioxidant activity spectral effect relationship were successfully established ；mangiferin（peak M 7）may be the main antioxidant substance of ethanol extract from A. asphodeloides . Timosaponin B Ⅱ（peak S 5），peak S 6 and peak S 4 may be the main antioxidant substance in acetone extract from A. asphodeloides .

Establishment of HPLC Fingerprint of Lonicera japonica and Study on Its Anti-inflammatory Spectrum-effect Relationship / 中国药房

OBJECTIVE：To establish fingerprint of Lonicera japonica ，and to study its anti-inflammatory spectrum-effect relationship. METHODS ：HPLC was adopted. The determination was performed on Diamonsil C 18 column with mobile phase consisted of 0.1％ formic acid solution-acetonitrile （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was 30 ℃，and detection wavelength was 238 nm. The sample size was 10 μL. Using chlorogenic acid as reference，HPLC fingerprint of 10 batches of L. japonica from different production areas was established according to TCM Chromatographic Fingerprint Similarity Evaluation System （2012 edition）. By comparing with reference substance ，chemical constituents corresponding to common peaks were identified ，and the similarity analysis was conducted. Acute and chronic inflammatory models of mice induced by xylene ，carrageenan and cotton ball were used to evaluate inhibition rate of 10 batches of L. japonica to ear，foot and granuloma swelling； the average value was calculated as the comprehensive pharmacodynamic index. The spectrum-effect relationship with HPLC fingerprint of L. japonica and anti-inflammatory effect was analyzed by grey relational analysis （GRA）and partial least squares regressiosn （PLSR）based on common peak area and comprehensive pharmacodynamic index . Chromatographic peaks with correlation＞0.7 and regression coefficient of PLSR model ＞0 were characteristic peaks. The percentage of peak areas of characteristic peaks to peak areas of common peak was calculated in 10 batches of L. japonica （e.g.“peak ratio ”）. RESULTS ： There were 25 common peaks in HPLC fingerprints of 10 batches of L. japonica ，with similarity of 0.775-0.994. Totally 9 peaks were confirmed ，i.e. rutin （peak 18），hyperoside（peak 20），isochlorogenc acid B （peak 22），galuteolin（peak 21），chlorogenc acid（peak 9），loganin（peak 10），neochlorogenic acid （peak 2），isochlorogenic acid C （peak 25），isochlorogenic acid A （peak 23）. All 10 batches of L. japonica had inhibitory effects on ear swelling ，foot swelling and granuloma ，with average inhibitory rate of 47.95％-56.52％. The correlation by GRA was peak 8＞12＞18＞16＞3＞11＞20＞22＞19＞21＞1＞9＞10＞13＞24＞14＞2＞ 17＞25＞23＞5＞4＞15，and all of correlations were greater than 0.7. The regression coefficient of PLSR for peaks 2，4，5，7，8， 10，12，13，14，15，16，17，18，20，21，22，24 were all greater than 0；those peaks were positively correlated with anti-inflammatory effect and were characteristic peaks except for peak 7； among them ，VIP values of peaks 5，8，10，16，18， 20，24 were greater than 1. The peak ratio of 10 batches of L. japonica was 58.61％-71.19％. CONCLUSIONS ：HPLC fingerprint of 10 batches of L. Japonica is successfully established. 10 batches of samples have similar components ，and the content of anti-inflammatory components is relatively high. The proportion of characteristic peaks to common peaks should not be less than 51.8％.

Establishment idea of Lonicerae Japonicae Flos fingerprint system based on Q-marker of Chinese materia medica / 中草药

The establishment of fingerprint system and the study of spectrum-effect relationship have become a very important development trend in quality evaluation of Chinese materia medica (CMM). It is based on the research of index components and can be used to evaluate the authenticity, quality and stability of crude drug, pieces and patent medicines of CMM. However, it is affected by many unstable factors and lacks of relevance to traditional Chinese medicine theories. Quality marker, in due course, provides an enlightening thought and way for the fingerprint system and spectrum-effect relationship studies. Taking Lonicerae Japonicae Flos as an example, this paper explores the researching ideas and establishes methods for fingerprinting research on CMM based on the characteristics of Q-markers.

Preliminary discovery of quality marker of herb Meconopsis integrifolia (non flower parts) based on spectrum-effect relationship / 中草药

Objective: To establish a study method for discovering quality marker of herb Meconopsis integrifolia (Non flower parts) based on spectrum-effect relationship for its quality control. Methods: UPLC fingerprints and UPLC-ESI-Q-TOF-MS/MS method were used to identify the chemical components of 10 batches of HMI extracts. The total antioxidant capacity, ABTS free radical scavenging ability, DPPH free radical scavenging ability and superoxide anion free radical suppression ability of 10 batches of sample were determined. Principal component analysis was used to screen antioxidant capacity index for HMI. Grey relational analysis was used to analyze the correlation between antioxidant capacity index and common peak of fingerprint spectrum. Then, quality marker could be preliminarily discovered by comprehensive analysis associated with antioxidant capacity index and efficacy-related constituents. Finally, the constituents were separated by HSCCC. Results: A total of 29 common peaks were identified from fingerprints of HMI extracts. 18 constituents were identified by UPLC-ESI-Q-TOF-MS/MS analysis, including 16 flavonoids and two alkaloids. DPPH scavenging ability was screened by principal component analysis which could reflect the antioxidant capacity of HMI extracts. Grey relational analysis showed that the correlations between the 29 common peaks and DPPH scavenging ability were all greater than 0.5. The quality markers of HMI were quercetin 3-O-β-D-glucopyrannosy-(1→6)-β-D-glucopyranoside and quercetin 3-O-[2'''-O-acetyl-β-D-galactopyranosyl-(1→6)-β-D-glucopyranoside] by comprehensive analysis. Among them, quercetin 3-O-[2'''-O-acetyl-β-D-galactopyranosyl-(1→6)-β-D- glucopyranoside] is a new compound. Conclusion: Study on the quality marker of HMI based on the spectrum-effect relationship is of great significance for elucidating the pharmacodynamic material basis, screening core quality marker related to medicinal effect, and ensuring the safety and rational application of traditional medicines.