Quality evaluation of Indigo Naturalis based on multi-component quantification combined with chemometrics / 中国药房

OBJECTIVE To evaluate the quality of Indigo Naturalis， and to provide reference for the quality control of Indigo Naturalis. METHODS UPLC-MS/MS method was used to determine the contents of 6 indole alkaloids （indigo， indirubin， isatin， tryptanthrin， indole and indole-3-carboxaldehyde） in Indigo Naturalis from different origins. Cluster analysis， principal component analysis and partial least squares-discriminant analysis （PLS-DA） were used to evaluate the quality of Indigo Naturalis from different origins. RESULTS The contents of indigo， indirubin， isatin， tryptanthrin， indole and indole-3-carboxaldehyde in Indigo Naturalis from different origins were 20 320.83-26 585.01， 1 327.69-3 102.25， 141.69-894.50， 2.17-5.27， 2.14-5.93 and 1.69-4.34 μg/g， respectively. The Indigo Naturalis from different areas were clustered into two categories by cluster analysis. Samples S1， S2， S4， S6， S7， S9 and S10 were clustered into category Ⅰ， and samples S3， S5， S8， S11 and S12 were clustered into category Ⅱ. Indigo Naturalis from different origins was evaluated with 3 principal components. The results showed that category Ⅰ sample scored higher and had better quality， while category Ⅱ sample scored lower and had worse quality. PLS-DA showed that indigo， indirubin， tryptanthrin and isatin were the main substances that reflected the quality difference of Indigo Naturalis. CONCLUSIONS The quality of Indigo Naturalis from different origins is different， and the quality of Indigo Naturalis of different batches from the same area is not stable. The quality evaluation method of Indigo Naturalis established in this paper is stable and reliable， which can provide a basis for the quality control of Indigo Naturalis.

Metabolomics-based Analysis of Toxicity Changes of Aconiti Lateralis Radix Praeparata-Zingiberis Rhizoma in Sinitang Before and After Compatibility / 中国实验方剂学杂志

ObjectiveTo explore the effect and toxicity change rule of Aconiti Lateralis Radix Praeparata（ALRP） and Zingiberis Rhizoma（ZR） before and after compatibility， and to reveal the compatibility connotation of them. MethodSixty SD rats were randomly divided into blank group， blank-ALRP group， blank-ALRP-ZR group， model group， model-ALRP group and model-ALRP-ZR group， the latter three groups were injected with adriamycin via tail vein to establish the model of heart failure， and the former three groups were injected with the same amount of physiological saline via tail vein. The effects of ALRP single decoction and ALRP-ZR mixed decoction on biochemical indexes and myocardial histopathological morphology of normal rats and model rats were compared. Metabolomics analysis was performed on rat serum samples， principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） were used to screen the differential metabolites between groups， and the differential metabolic pathways were analyzed. Combined with network pharmacology technology， the metabolites and their associated targets and pathways related to enhancing anti-heart failure efficacy and reducing cardiotoxicity were screened before and after the compatibility of ALRP and ZR， the screened representative pathways were verified by Western blot. ResultCompared with the blank group， the model group showed significant increases in the contents of brain natriuretic peptide（BNP）， creatine kinase（CK）， lactate dehydrogenase（LDH） and cardiac troponin（cTn）-T（P<0.01）， the blank-ALRP group showed obvious increases in CK， LDH， and cTn-T contents（P<0.05， P<0.01）， while the normal-ALRP-ZR group showed a significant increase in CK content（P<0.01）. Compared with the blank-ALRP group， the blank-ALRP-ZR group showed a obvious decrease in LDH content（P<0.05）， and pathological sections showed that both decoctions could lead to myocardial histopathological damage in normal rats. Compared with the model group， the model-ALRP-ZR group showed obvious decreases in BNP， CK， LDH and cTn-T contents（P<0.05， P<0.01）， and the model-ALRP group showed obvious decreases in BNP， LDH and cTn-T contents（P<0.05， P<0.01）. Compared with the model-ALRP group， the model-ALRP-ZR group showed a significant decrease in CK content（P<0.01）， and both decoctions could improve the pathological morphology of myocardial tissue in the model rats. Metabolomics results showed that ALRP single decoction and ALRP-ZR mixed decoction could recover 422 and 459 metabolites in model rats， respectively. And the metabolic disruption of ALRP-ZR mixed decoction on normal rats was weaker than that of ALRP single decoction. The results of network pharmacological association analysis showed that in the aspect of ZR enhancing the anti-heart failure efficacy of ALRP， 3 metabolites such as deoxyuridylic acid were correlated to 56 metabolites， 82 targets and 13 pathways， including calcium signaling pathway， renin secretion， renin-angiotensin system， etc. In the aspect of ZR reducing the cardiotoxicity of ALRP， 3 metabolites such as tyrosol were associated with 24 metabolites， 55 targets and 14 pathways， including adrenergic signaling in cardiomyocytes and carbon metabolism and so on. Western blot results showed that the expression of angiotensin-converting enzyme（ACE）， angiotensin-converting enzyme 2（ACE2） and angiotensin Ⅱ（Ang Ⅱ） in myocardial tissues of rats from the model group was significantly elevated by comparing with the blank group（P<0.01）. Compared with the model group， the model-ALRP group and the model-ALRP-ZR group showed significantly decreased expression of ACE， ACE2 and Ang Ⅱ（P<0.01）. Compared with the model-ALRP group， the expression of ACE2 and AngⅡ was significantly decreased in the model-ALRP-ZR group. Compared with the blank group， the expression of extracellular signal regulated kinase（ERK）， protein kinase B（Akt） and cTn-I3 was significantly elevated in the blank-ALRP group and blank-ALRP-ZR group（P<0.01）. Compared with the blank-ALRP group， the blank-ALRP-ZR group showed decreased expression of ERK， Akt and cTn-I3， but there was no statistical significance. ConclusionTo a certain extent， the combination of ALRP and ZR shows synergistic relationship under pathological state， and attenuated effect of compatibility under normal physiological state， and the pharmacodynamic characteristics and compatibility relationship of ALRP and ZR are closely related to the physiological state.

Comparison of the Contents of 14 Inorganic Elements in Cyperus rotundus from Different Origins Based on Stoichiometry / 中国药房

OBJECTIVE：To establish the method for the content determination of inorganic elements in Cyperus rotundus ，and to compare the contents of 14 kinds of inorganic elements in C. rotundus from different producing areas ，and to provide theoretical basis for its quality control and high quality resources development . METHODS ：The samples were processed by microwave digestion，and ICP-MS method was used to determine the contents of Na ，Mg，K，Ca，Mn，Fe，Ni，Cu，Zn，As，Se，Sr，Cd and Pb. SPSS 23.0 software were used for principal component analysis （PCA）and cluster analysis. RESULTS ：The average contents of above 14 kinds of inorganic elements in C. rotundus from different producing areas were 168.62，753.71， 6 938.33，24.31，14.69，197.77，0.60，2.43，26.89，0.21，0.06，5.64，0.05，0.32 mg/kg，respectively. The results of PCA showed that the cumulative variance contribution rate of the first four principal components was 86.203％，which could reflect most of the information of the original data. C. rotundus from Shandong ，Jiangxi，Shanxi，Hubei and Yunnan ranked the top five places in terms of comprehensive score of inorganic element contents. The results of cluster analysis showed that the samples from 9 producing areas were clustered into 5 categories，showing the characteristics of clustering by producing area. From the perspective of inorganic elements ，the quality of C. rotundus from East China ，Central China ，North China and Southwest China was better than that from South China. CONCLUSIONS ：Essential trace elements like Na ，Mg，K，Ca，Mn，Fe，Cu，Zn，Sr are rich in C. rotundus，and there are small amounts of Ni ，As，Se，Cd，Pb elements in it. The contents difference of inorganic elements in C. rotundus from different origins may related to the geographical area it belongs to.

Comparative Study on the Contents of 4 Components in the Volatile Oil of Cyperus rotundus from Different Origins Based on HPLC Method and Multivariate Statistical Analysis / 中国药房

OBJECTIVE： To estab lish the method for simultaneous determination of the contents of cyperotundone ， nootkatone，α-cyperone and aristolone in the volatile oil of Cyperus rotundus ，compare the content differences of 4 components in C. rotundus samples from different origins ，and to provide reference for germplasm screening ，development and utilization of the medicinal material. METHODS ：The volatile oil was extracted from 46 batches of C. rotundus from 12 origins. The contents of cypermenone，nootkatone，α-cyperone and aristolone in volatile oil were determined by HPLC. The determination was performed on Kromasil C 18 column with mobile phase consisted of methanol-water （68∶32，V/V）at the flow rate of 1.0 mL/min；the column temperature was 30 ℃；the detection wavelength was set at 242 nm；the sample size was 20 μL. Using the contents of above 4 components as evaluation indexes ，radar image analysis ，cluster thermal map analysis and principal component analysis were performed for comparing the quality of C. rotundus from different origins. RESULTS ：The results of content determination methodology investigation met relevant requirements ；the total contents of 4 components in volatile oil from C. rotundus from different origins ranged from 136.986 4 to 538.832 1 mg/g，of which the total content of samples from Yunnan was the highest （the average value was 476.059 2 mg/g）. Radar image analysis results showed that the whole contour in the 4 origins of Guangdong ， Jiangxi，Guangxi and Yunnan was large relatively and better balanced ，among which the samples from Yunnan had the largest overall contour and the best balance. The cluster thermal map analysis results showed that the samples from 12 origins could be grouped into 2 categories，the first category was from Hubei ，Jiangxi，Yunnan，Sichuan，Guangdong，Shandong，Henan and Shaanxi；the second category was from Guangxi ，Shanxi，Anhui and Hainan ；the quality of samples from the first category were better than that of samples from the second category. The principal component analysis results showed that the cumulative contribution rate of the first three principal components was 96.1％，and the samples from 12 origins were mainly clustered into two categories ，which was consistent with the results of cluster thermal map analysis. CONCLUSIONS ：Established HPLC method can be used for simultaneous determination of cypermenone ，nootkatone，α-cyperone and aristolone in volatile oil of C. rotundus from different origins. Among the samples from 12 origins，the quality of medicinal material from Yunnan is better.

Injectable thermo-responsive nano-hydrogel loading triptolide for the anti-breast cancer enhancement

The clinical application of triptolide (TPL) in tumor therapy has been greatly limited by its toxicity and inefficient delivery. Herein, a localized and sustained-release thermo-sensitive hydrogel was developed for the intra-tumor administration of TPL. Based on the amphiphilic structure of poly (

Study on the Optimization of the Extraction Technology of Shengmaiyin Polysaccharide and Its Regulation Effects on Intest- inal Function of Spleen Deficiency Model Rats / 中国药房

OBJECTIVE： To optimize the extraction technology of Shengmaiyin polysaccharide， and to investigate the regulation effects of Shengmaiyin and its polysaccharide on intestinal function of spleen deficiency model rats. METHODS： The contents of polysaccharide were determined by phenol-sulfuric acid method， and the extraction rate of polysaccharide was calculated. Using extraction rate of Shengmaiyin polysaccharide as investigation index， singel factor and orthogonal tests were used to optimize material-liquid ratio， extraction time， extraction temperature and extraction times of Shengmaiyin polysaccharide. Validation test was also conducted. Totally 80 male SD rats were randomly divided into blank group， model group， Shengmaiyin low-dose， medium-dose and high-dose groups （350， 700， 1 400 g/L， by crude drug）， Shengmaiyin polysaccharide low-dose， medium-dose and high-dose groups （24.5， 49， 98 g/L， by crude drug）， with 10 rats in each group. Except for blank group， other groups were given Rheum palmatum water decoction 10 mL/kg to induce spleen deficiency model， once a day， for consecutive 15 d. Since the 16th day， blank group and model group were given isovolumic water intragastrically， while other groups were given corresponding drugs， once a day， for consecutive 10 d. The general status of rats and body weights were recorded in each group. The serum contents of D-xylose， gastrin （GAS） and vasoactive intestinal peptide （VIP） were detected by phloroglucinol method or ELISA. RESULTS： The optimal extraction technology of Shengmaiyin polysaccharide was material-liquid ratio 1 ∶ 10（g/mL）， extraction time 45 min， extraction temperature 80 ℃， extracting for 1 time. Results of validation test showed that extraction rates of the polysaccharide in 3 times were 7.43％， 7.64％， 7.80％ （RSD＝1.01％， n＝3）. After modeling， except for blank group， other groups suffered from loose stools， thin body and reduced food intake， and the body weight and serum level of D-xylose were decreased significantly compared with blank group （P＜0.01）. After last medication， above symptoms of administration groups were improved to different extents. Except for model group， body weight and serum contents of D-xylose in other groups were increased significantly than those before modeling or before medication （P＜0.05 or P＜0.01）. Compared with blank group， body weight and serum content of GAS were decreased significantly in model group， while serum content of VIP was increased significantly （P＜0.01）. Compared with model group， body weight of Shengmaiyin medium-dose group and Shengmaiyin polysaccharide low-dose and high-dose groups， serum contents of D-xylose and GAS in Shengmaiyin medium-dose and high-dose groups and Shengmaiyin polysaccharide low-dose and medium-dose groups were increased significantly， while serum contents of VIP in Shengmaiyin groups and Shengmaiyin polysaccharide low-dose and medium-dose groups were all decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： The optimized extraction technology of Shengmaiyin polysaccharide is stable and feasible. Shengmaiyin and its polysaccharide contribute to the recovery of intestinal function of spleen deficiency model rat， the effects of which may be associated with the secretion regulation of GAS and VIP.

A novel quantified bitterness evaluation model for traditional Chinese herbs based on an animal ethology principle

Traditional Chinese herbs (TCH) are currently gaining attention in disease prevention and health care plans. However, their general bitter taste hinders their use. Despite the development of a variety of taste evaluation methods, it is still a major challenge to establish a quantitative detection technique that is objective, authentic and sensitive. Based on the two-bottle preference test (TBP), we proposed a novel quantitative strategy using a standardized animal test and a unified quantitative benchmark. To reduce the difference of results, the methodology of TBP was optimized. The relationship between the concentration of quinine and animal preference index (PI) was obtained. Then the PI of TCH was measured through TBP, and bitterness results were converted into a unified numerical system using the relationship of concentration and PI. To verify the authenticity and sensitivity of quantified results, human sensory testing and electronic tongue testing were applied. The quantified results showed a good discrimination ability. For example, the bitterness of Coptidis Rhizoma was equal to 0.0579 mg/mL quinine, and Nelumbinis Folium was equal to 0.0001 mg/mL. The validation results proved that the new assessment method for TCH was objective and reliable. In conclusion, this study provides an option for the quantification of bitterness and the evaluation of taste masking effects.

Prediction of the anti-inflammatory mechanisms of curcumin by module-based protein interaction network analysis

Curcumin, the medically active component from Curcuma longa (Turmeric), is widely used to treat inflammatory diseases. Protein interaction network (PIN) analysis was used to predict its mechanisms of molecular action. Targets of curcumin were obtained based on ChEMBL and STITCH databases. Protein-protein interactions (PPIs) were extracted from the String database. The PIN of curcumin was constructed by Cytoscape and the function modules identified by gene ontology (GO) enrichment analysis based on molecular complex detection (MCODE). A PIN of curcumin with 482 nodes and 1688 interactions was constructed, which has scale-free, small world and modular properties. Based on analysis of these function modules, the mechanism of curcumin is proposed. Two modules were found to be intimately associated with inflammation. With function modules analysis, the anti-inflammatory effects of curcumin were related to SMAD, ERG and mediation by the TLR family. TLR9 may be a potential target of curcumin to treat inflammation.

Chemical ingredient analysis of sediments from both single Radix Aconiti Lateralis decoction and Radix Aconiti Lateralis - Radix Glycyrrhizae decoction by HPLC-MS / 药学学报

The chemical ingredients of sediments from single Radix Aconiti Lateralis (Fuzi, in Chinese) decoction and Radix Aconiti Lateralis-Radix Glycyrrhizae (Gancao, in Chinese) decoction were evaluated separately by HPLC-MS for exploring their ingredients difference and discussing the mechanism of toxicity reduction of Fuzi-Gaocao herb-pair, even providing the experimental basis for extracting and purifying process of TCM preparations containing Fuzi-Gancao herb-pair. The sediments samples from single Fuzi decoction (FD) and Fuzi-Gaocao decoction (FGD) were prepared separately firstly. Then these samples were analyzed in the same chromatographic and mass spectrometry condition, estimating their HPLC-MS fingerprint and identifying their compounds by Q-TOFMS. As a result, 28 compounds in sediments samples from FD were obtained, among which 25 compounds and structures were confirmed. Besides, 36 compounds in sediments samples from FGD sample were obtained including 34 confirmed compounds and structures, among which there were 11 compounds from Gancao and 25 compounds from Fuzi. However, according to these confirmed compounds, alkaloids compounds in sediments samples from FGD were significantly different from that in sediments samples from FD. This result showed the fact that there would be some influences on alkaloid compounds in decoction after the combination of Gancao. In this study, the experiment basis of the mechanism of toxicity reduction of Fuzi-Gaocao herb-pair is provided. It also showed that much attention should be paid to the fact that there were great amount of alkaloid compounds, the effective compounds, in the sediments for which a comprehensive utilization in clinic and pharmaceutical process should be noted.

Study on stability of major pharmaceutical ingredients of vladimiriae radix before and after being roasted in artificial gastric juice, artificial intestinal juice and isolated rat gastric, intestinal or colonic incubation juice / 中国中药杂志

<p><b>OBJECTIVE</b>To study the stability of costunolide (COS) and dehydrocostus lactone (DEH) of Vladimiriae Radix before and after being roasted in artificial gastric juice, artificial intestinal juice and isolated rat gastric, intestinal or colonic incubation juice.</p><p><b>METHOD</b>The HPLC method was used for the determination of the mass concentration of COS and DEH Vladimiriae Radix before and after being roasted artificial gastric juice, artificial intestinal juice and isolated rat gastric, intestinal or colonic incubation juice. The samples were incubated with isolated rat stomach, small intestine; colon was used to study physical adsorption, absorption or degradation parameters.</p><p><b>RESULT</b>COS of Vladimiriae Radix before or after being roasted was unstable in artificial gastric juice, with the average degradation constants as 0.758 0 and 0.531 1. Having been roasted, it showed an increasing stability with a significant difference (P < 0.01). Both of COS and DEH of Vladimiriae Radix before or after being roasted showed high adsorption, uptake or degradation (2 h), and it had significant difference between different parts.</p><p><b>CONCLUSION</b>COS was unstable in artificial gastric juice (unprocessed Vladimiriae Radix has a higher degradation rate). Isolated rat stomach, small intestine, colon can adsorb, take, degrade COS and DEH of Vladimiriae Radix before or after roasting process obviously and differently. It provides basis for studies on the absorption mechanisms of effective ingredients of Vladimiriae Radix before and after being roasted.</p>

Intestinal lymphatic transport of breviscapine orally administered in rat / 药学学报

Double cannulation model of conscious rat allowing simultaneous collection of mesenteric lymph and jugular venous blood was established to investigate the intestinal lymphatic transport of breviscapine orally administered in rat. The concentrations of breviscapine in plasma and lymph were determined by HPLC. The pharmacokinetics of breviscapine after oral and intravenous administration was evaluated in the conscious rat model. It was observed that scutellarin distributed from blood circulation to lymphatic system after intravenous injection. The cumulative lymphatic transport amount within 12 h was (2.78 +/- 0.25) microg, equivalent to 0.0792% of intravenous dose. After oral administration of scutellarin to double-cannulation rats, the cumulative lymphatic transport amount within 12 h was (0.92 +/- 0.08) microg, equal to 0.0083% of oral dose. The absolute bioavailability of breviscapine orally administered to double-cannulation rats was 4.91%, indicating that scutellarin was mainly absorbed into the bloodstream through the portal vein. Lymphatic transport of scutellarin appears to reflect high affinity for the lymph lipoproteins to chylomicron. This study provided a biopharmaceutics basis for developing oral lipid delivery system for the promotion of intestinal lymphatic transport to improve oral bioavailability of breviscapine.

In vitro balanced sustained-release of Panax notoginseng saponins controlled with various matrix materials / 药学学报

To explore the influence of matrix materials in complicate ingredients on traditional Chinese medicine and investigate the excipients selection model based on balanced release characteristics of multicomponents, the influence of HPMC (K4M, K15M, K100M) and Carbomer (934P, 971P, 974P) was illustrated by testing in vitro release of ginsenoside-Rg1, ginsenoside-Rb1 and notoginsenoside-R1 in Panax notoginseng saponins (model drug, PNS). According to in vitro release results of PNS matrix tablets in water and artificial intestinal juice, the release curves were analyzed with Peppas equation and simulating factor (f). Significant differences in k value and n value among ginsenoside-Rg1, ginsenoside-Rb1 and notoginsenoside-R1 existed in various formulations. The release behaviors from various excipients could be described with Non-Fickian transport or super Case II transport pattern. The f2 values for ginsenoside-Rg1, ginsenoside-Rb1 and notoginsenoside-R1 in 971P matrix tablet containing 30% Carbomer 971P were 74.91, 53.45, 57.89 in water and 79.35, 55.51, 51.89 in artificial intestinal juice, respectively. The release profiles fit for the regulation of FDA. The result revealed that the balanced release rates of Rg1, Rb1 and R1 in 971P matrix tablet were obtained.

Effect of Baked Radix Vladimiriae on the Spasm of in Vitro Intestinal Muscle in Rabbit / 中国药房

OBJECTIVE:To explore the spasmolytic effect of baked Radix Vladimiriae on in vitro intestinal muscle in rabbits. METHODS:Magnus in vitro intestinal muscle accumulative dose method was adopted to observe the dose-effect relationship of petroleum ether parts of baked Radix Vladimiriae with in vitro intestinal muscle of rabbits and its impact on the spasm of rabbit intestinal muscle induced by acetylcholine, histamine phosphate and Bacl2. RESULTS:There was a certain dose-effect relationship between petroleum ether parts of baked Radix Vladimiriae and spasmolytic mechanism of rabbit in vitro intestine. It showed the effect of acetylcholine, histamine phosphate and Bacl2. CONCLUSION:The inhibition effect of petroleum ether parts of baked Radix Vladimiriae on normal in vitro intestinal muscle increase as long as dose increase. The inhibition mechanism may be associated with acetylcholine competing with histamine to block M receptor and H1 receptor as well as to inhibit excitability of intestinal muscle.

Quality standard for Nabao Capsule / 中成药

AIM:To establish the quality standard for Nabao Capsule(Radix Astragali,Radix Angelicae sinensis,Rhizoma Corydalis,Herba Hedyotis,etc.).METHODS:Radix Angelicae sinensis,Rhizoma Corydalis,Herba Hedyotic and Pendulous Monkshood Root in Nabao Capsule were identified by TLC.Astragaloside Ⅳ was determined by HPLC.RESULTS:The characteristic identification by TLC was distinct and highly specific,the quantitative evaluation of thymol had the linear range of 1-10 ?g.The average recovery was 99.05% and RSD was 2.48%.CONCLUSION:The method is reliable,accurate and specific,and can be used for the quality control of Nabao Capsule.