Comprehensive quality consistency evaluation strategy and analysis of compound danshen tablet.

The compositions of traditional Chinese medicines are extremely complex,as a result, exploring consistent quality is demanded and challenging. Quality consistency of products obtained from the same manufacturer has received little attention. The strategy of quality consistency evaluation (QCE) has been proposed as a novel method for quality control of Traditional Chinese Medicine Patent Prescription (TCMPP). This study aimed to establish a comprehensive QCE strategy for Compound Danshen Tablet (CDT). High Performance Liquid Chromatography-Diode Array Detector and Gas Chromatography-Mass Spectrometry were separately applied to determinate the content of seven and two index components, which representing the quality actuality of different raw medicines. The dissolution test was designed to obtain the dissolution ratios of CDT samples. QCE can provide the intra-batch content consistency difference (PA), inter-batch content consistency difference (PB), and dissolution ratio consistency difference (PR) values. The consistency of CDT samples from 15 different manufacturers (75 batches) was evaluated by principal component analysis (PCA), which showed that the total content (nine index components) of the 75 batches of samples obtained from 15 manufacturers ranged from 22.11 to 38.45 mg·tablet-1. The dissolution ratios ranged from 74.8% to 116.4%. The PA values of 15 manufacturers ranged from 2.4% to 12.2%, and the PB (11.1-45.1%) values were higher than the PA values. The PR values reflecting the various dissolution ratios in vitro ranged from 8.1% to 57.5%. The three consistency factors were ranked by PCA, and products of the 15 manufacturers were classified into three categories. The PA, PB, and PR values provided a comprehensive and effective approach for monitoring the quality consistency of CDT and can serve as an example of QCE for other TCMPP.

Anti-hyperlipidemic effects of the compound Danshen tablet: roles of antioxidation, anti-inflammation, anticoagulation, and anti-apoptosis.

BACKGROUND: Hyperlipidemia could cause some serious harm to human health diseases, such as atherosclerosis, coronary heart disease. This study sought to investigate the effects of the compound Danshen tablet (CDT) on hyperlipidemia induced by a high-fat diet in ApoE-/- mice and related antioxidation, anti-inflammation, anticoagulation, and anti-apoptosis mechanisms. METHODS: The control group (Group 1) comprised 15 male C57BL/6N mice, and the other 5 groups (Groups 2-6) comprised 75 male ApoE-/- mice. These 75 mice were randomly divided into 1 of the following 5 groups: Group 2, a model group; Groups 3-5, the CDT groups, each of which was administered 375, 750, or 1,500 mg/kg of CDT; and Group 6, an atorvastatin group, which was administered 5.2 mg/kg of atorvastatin. All the mice were fed a high-fat diet for 16 weeks and intragastrically administered with CDT or atorvastatin once a day according to their body weight. After 16 weeks, serum was collected, the aorta was isolated, and blood lipid levels were detected. An enzyme-linked immunosorbent assay was used to detect the serum levels of 4-hydroxynonenal (4-HNE), 8-hydroxy-2'-deoxyguanosine (8-OHdG), intercellular adhesion molecule 1 (ICAM-1), monocyte chemoattractant protein 1 (MCP-1), thromboxane B2 (TXB2), tissue plasminogen activator (tPA), and plasminogen activator inhibitor-1 (PAI-1). The thickness of the aortic wall was measured by ultrasonography. Atherosclerotic plaque and endothelial cell apoptosis in the aortic root were evaluated using oil red O staining and terminal dUTP nick-end labeling (TUNEL) assays, respectively. RESULTS: A comparison of mice in the CDT group and mice in the model group showed that CDT significantly inhibited mice's weight gain. CDT reduced the levels of the inflammatory factor ICAM-1 and the oxidative damage molecule 4-HNE. In the coagulation system, CDT significantly increased tPA levels and reduced TXB2 and PAI-1 levels. Ultrasonography showed that CDT increased the thickness of the aortic wall. The oil red O staining results revealed that CDT significantly ameliorated lipid accumulation in the aortic valve. TUNEL assays indicated that CDT reduced the number of TUNEL-positive cells in the aortic valve. CONCLUSIONS: CDT has a certain protective effect on hyperlipidemia. The mechanism of CDT may be related to antioxidation, anti-inflammation, anticoagulation, and anti-apoptosis.

Transcriptomics and metabolomics reveal the cardioprotective effect of Compound Danshen tablet on isoproterenol-induced myocardial injury in high-fat-diet fed mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Compound Danshen tablet, an herbal preparation consisting of salviae miltiorrhizae, notoginseng and borneolum, is extensively employed clinically to treat angina pectoris, coronary arteriosclerosis and significantly improve microcirculation. AIM OF THE STUDY: To reveal the potential underlying cardioprotective mechanism(s) in isoproterenol-induced myocardial injury in high-fat-diet fed mice. MATERIALS AND METHODS: Cardiac transcriptomics was analyzed by Illumina mRNA-Seq sequencing. The restored cardiovascular diseases (CVD)-related genes by Compound Danshen tablet were validated by quantitative real time polymerase chain reaction (qRT-PCR). Furthermore, Cardiac metabolomics were also performed using gas chromatography-mass spectrometry. RESULTS: From the transcriptomics study, we found the levels of 24 up-regulated and 44 down-regulated genes in the control compared to model groups. Among them, seven gene levels were restored by treatment of Compound Danshen tablet. Four CVD-related genes at the mRNA level (Sprr1a, Ppp1r3c, Bmp10 and Hspa1b) were validated successfully by qRT-PCR. From the metabolomics study, 37 differentially expressed metabolites were identified between the control and model groups. Among them, 21 metabolites were restored by treatment of Compound Danshen tablet. These altered metabolites are involved in glucose metabolism, fatty acid metabolism and amino acid metabolism. CONCLUSION: These genes and metabolites might provide clues for further molecular mechanistic study of Compound Danshen tablet.

Bioassay method for Compound Danshen Tablets based on a platelet aggregation inhibition test / 药学学报

A bioassay method for inhibiting platelet aggregation in vitro was established to quantify the pharmacological effects of Compound Danshen Tablets and support its quality control. The inhibition of platelet aggregation in rabbit plasma in vitro by Compound Danshen Tablets was used as the experimental system. The titer was calculated by using the method of dose-response parallel lines. As a result, a bioassay for the inhibition of platelet aggregation in vitro by Compound Danshen Tablets was established. Linearity was good in the concentration range of 0.128 g·mL-1 to 0.205 g·mL-1, and the titer of standard Compound Danshen tablets was 7 659 U·g-1 according to the titer definition. This in vitro assay was simple, reliable, reproducible and convenient. The activity of Compound Danshen Tablets in inhibiting platelet aggregation was quantified by potency assay and the quality of different batches was evaluated. The method can be applied for the quality control of Compound Danshen Tablets.

[Determination of salvianolic acid B and borneol in compound Danshen tablet by near-infrared spectroscopy and establishment of dependency model].

To determine the contents of salvianolic acid B and borneol in compound Danshen tablet (composed of Salviae Miltiorrhizae Radix et Rhizome, Notoginseng Radix et Rhizome and Borneolum Syntheticum) by near-infrared spectroscopy (NIR) and to establish a dependency model for rapid quantitative analysis. NIR data of 74 batches of compound Danshen tablet from different companies were collected; the contents of salvianolic acid B and borneol were determined by using high performanceliquid chromatography (HPLC) and gas chromatography (GC) respectively to establish the dependency model for salvianolic acid B and borneol. The results showed that the best waveband for salvianolic acid B was 10 846.2-10 013, 9 195.3-8 362.2, 6 719.1-4 242.8 cm⁻¹; root-mean-squares error of cross-validation (RMSECV), coefficient of determination R² and regression point displacement (RPD) of salvianolic acid B were 1.72 mg·g⁻¹, 91.05% and 7.93 mg·g⁻¹, respectively. While the best wavebands, RMSECV, R² and RPD of borneol were 10 846.2-5 060.5 cm⁻¹, 1.2 mg·g⁻¹, 96.11% and 6.71 mg·g⁻¹ï¼Œrespectively. The relative error of the established model as validation results for validation set samples was 2.67% and 4.64%， respectively.With the good predictability, the models can be applied to the real time monitoring and quality control of compound Danshen tablet.

Determination of salvianolic acid B and borneol in compound Danshen tablet by near-infrared spectroscopy and establishment of dependency model / 中国中药杂志

To determine the contents of salvianolic acid B and borneol in compound Danshen tablet (composed of Salviae Miltiorrhizae Radix et Rhizome, Notoginseng Radix et Rhizome and Borneolum Syntheticum) by near-infrared spectroscopy (NIR) and to establish a dependency model for rapid quantitative analysis. NIR data of 74 batches of compound Danshen tablet from different companies were collected; the contents of salvianolic acid B and borneol were determined by using high performanceliquid chromatography (HPLC) and gas chromatography (GC) respectively to establish the dependency model for salvianolic acid B and borneol. The results showed that the best waveband for salvianolic acid B was 10 846.2-10 013, 9 195.3-8 362.2, 6 719.1-4 242.8 cm⁻¹; root-mean-squares error of cross-validation (RMSECV), coefficient of determination ² and regression point displacement (RPD) of salvianolic acid B were 1.72 mg·g⁻¹, 91.05% and 7.93 mg·g⁻¹, respectively. While the best wavebands, RMSECV, ² and RPD of borneol were 10 846.2-5 060.5 cm⁻¹, 1.2 mg·g⁻¹, 96.11% and 6.71 mg·g⁻¹，respectively. The relative error of the established model as validation results for validation set samples was 2.67% and 4.64%， respectively.With the good predictability, the models can be applied to the real time monitoring and quality control of compound Danshen tablet.

Feasibility study of QAMS for quality control on three kinds of related preparations of Salvia miltiorrhiza / 中草药

Objective: To establish a quantitative analysis method of multi-components by single marker (QAMS) for the simultaneous determination of four tanshinones [tanshinone IIA (TIIA), dihydrotanshinone I (DTI), cryptotanshinone (CT), and tanshinone I (TI)], and to explore its accuracy and feasibility on quality evaluation of traditional chinese medicine preparations about three kinds of related preparations of Salvia miltiorrhiza. Methods: An HPLC method was used to calculate relative correction factors (fi/s) between other three components with TIIA. The component positioning methods were compared including retention time difference value, relative retention value, and linear regression methods. Meanwhile, the contents of tanshinones in different preparations of S. miltiorrhiza were determinated by the external standard method (ESM), including Danshen Tablet, Compound Danshen Tablet, and Guanxin Danshen Capsule. Measured content value was compared with calculated content value to evaluate the accuracy of QAMS method. Results: Within a certain range, the fi/s of DTI, TI, and CT to TIIA were 0.734, 0.916, and 0.774, respectively, with the good repeatability in different experimental conditions. The fi/s between other three components with TIIA was accurate and reliable, and there was no significant difference between QAMS and ESM method. Besides, the relative retention value method could be used to accurately locate the measuring component. The relative retention values of DTI, TI, and CT to TIIA were 0.314, 0.518, and 0.561, respectively. The contents of three kinds of S. miltiorrhiza preparations showed there were some differences in the intrinsic quality of the products from different factories, even from different batches of the same manufacturer. Conclusion: The QAMS method could be used for quality control about related preparations of S. miltiorrhiza and provide references for quality evaluation of other preparations. At the same time, the above three kinds of traditional Chinese medicine preparations should be further strengthened the control and supervision.

Efficacy Observation of Recurrent Oral Ulceration with Combination of Zhibai Dihuang Pills and Compound Danshen Tablets / 中国药房

OBJECTIVE:To observe clinical efficacy of recurrent oral ulceration with combination of Zhibai dihuang pills and Compound danshen tablets. METHODS:110 patients with recurrent oral ulceration were selected and divided into control group and tri-al group with 55 cases in each group. Control group individually received vitamin C 0.2 g/time+vitamin B2 10 mg/time+levamisole 45 mg/time,three times a day,Cetylpyridinium chloride collutory,15 ml/time,for 1 min,3 times a day. Trial group received Zhibai di-huang pills orally,8 pills/time,Compound danshen tablets,3 tablets/time,3 times a day. A treatment course lasted for 10 days. Both groups received 2 courses of treatment. Therapeutic efficacy,VAS and relapse were observed and compared between 2 groups. RE-SULTS:The cure rate(80.00%)and total effective rate(94.55%)of trial group were higher than those of control group(47.27%, 74.55%),with statistical significance(P<0.05). The VAS score of trial group were significantly lower than that of control group after 3 and 7 days of treatment,with statistical significance(P<0.05). After 2,6,10 and 12 months of treatment,the recurrence rates of tri-al group were 0,7.27%,9.09% and 16.36%,which were all significantly lower than those of control group(18.18%,34.55%, 47.27%,56.36%),with statistical significance(P<0.05). CONCLUSIONS:Combination of Zhibai dihuang pills and Compound danshen tablets has definite effect on recurrent oral ulcer,showing high cure rate and not easily recur.

Study of the protective mechanisms of Compound Danshen Tablet (Fufang Danshen Pian) against myocardial ischemia/reperfusion injury via the Akt-eNOS signaling pathway in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Compound Danshen Tablet (CDT), an herbal preparation consisting of Salvia miltiorrhiza (Radix and rhizome of Salvia miltiorrhiza Bge.), Notoginseng (Radix and rhizome of Panax notoginseng (Burk.) F. H. Chen) and Borneolum Syntheticum, is widely used to improve coronary heart disease, cardiac angina and atherosclerosis in clinic in Asia and Pacific Ocean area, especially in China. AIM: The study is to research the protective mechanisms of Compound Danshen Tablet (CDT) against myocardial ischemia/reperfusion (MI/R) injury via the Akt-eNOS signaling Pathway in rats. MATERIALS AND METHODS: Rats were randomized into 7 groups: Sham group; Model group; Low-Dose CDT group (0.315 g/kg/d, i.g); Middle-Dose CDT group (0.63 g/kg/d, i.g); High-Dose CDT group (1.26 g/kg/d, i.g); Compound Danshen Dripping Pills (CDDP) group (0.0945 g/kg/d, i.g); Sulfotanshinone Sodium Injection (Tan II A) group (5 mg/kg/d, i.m). After the administration, the hearts of the rats were subjected to 30 min of coronary artery occlusion and 2h of reperfusion except the Sham group rats. RESULTS: CDT significantly decreased infarct size, apoptosis, caspase-3 protein expression, MDA level in myocardial tissues, the activities of serum CK, LDH and cTnI; on the contrary, it increased p-Akt, p-eNOS, bcl-2 protein expression, the activities of SOD and tissue LDH, and the level of NO. CONCLUSIONS: CDT can protect cardiomyocytes against MI/R injury and inhibits apoptosis in rats by activating Akt-eNOS signaling pathway.

Comparative Studies on Pharmacological Action of Compound Danshen Aerosol and Tablet / 中药新药与临床药理

Compound Danshen aerosol remarkably increased hypoxia tolerance after isoproterenol injection and prolonged survival time in mice. The electrocardiogram of rats and rabbits with pituitrin-induced ischemia was significantly improved. The aerosol also decreased the elevation of lactic dehydrogenase in rats with experimental myocardial infarction.

Determination of tanshinone Ⅱ_A and cryptotanshinone in Compound Danshen Tablet by RP-HPLC / 中成药

AIM:To set up a determination of tanshinone Ⅱ_A and cryptotanshinone in Compound Danshen Tablet(Radix Salviae Miltiorrhizae, Radix Notoginseng, Borneolum Syntheticum) by RP-HPLC. METHODS:Two components could be separated through YWG-C_~18 column with methanol-water (75∶25 by volume) as a mobile phase. The flow rate was 1.0 mL?min~-1 , and the detection wavelength was at 270 nm. RESULTS: The linear range of tanshinone Ⅱ_A and cryptotanshinone were 2.5-50.0 ?g?mL~-1 and 0.625-12.5 ?g?mL~-1 , respectively. The average recovery was 99.76% with RSD 0.92% for tanshinone Ⅱ_A and 98.34% with RSD 0.71% for cryptotanshinone, respectively. CONCLUSION: The method is simple, accurate and rapid with good reproducibility. It can be used for the quality control of Compound Danshen Tablet.

Rapid nondestructive quality control of Compound Danshen Tablet by AOTFnear infrared spectroscopy / 中成药

AIM:A new method to rapidly non-destructive quality control Compound Danshen Tablet was developed by portable acousto-optic tunable filter (AOTF)-near infrared (NIR) spectroscopy. METHODS:A portable AOTF-NIR spectrometer was used to test 215 different batches of Compound Danshen Tablet collected from fifty-six pharmaceutical factories. The samples were analyzed by the Unscrambler program using principal component analysis(PCA). Quantitative analytical models invoking the partial least squares (PLS) algorithm simultaneously predicted the contents of four major constituents in Compound Danshen Tablet:salvianolic acid B,tanshinoneⅡA,water extract and alcohol extract. RESULTS:The qualitative analysis models accurately identified the differences between Compound Danshen Tablet and other reference substances. The results of external validation showed by the significant t-test that no significant difference between predictive and the chemical value,RSD was equal to 0. 990 9,0. 991 3,0. 964 0 and 1. 73% ,2. 08% ,3. 32% ,1. 32% ,respectively. CONCLUSION:The method of rapid non-destructive analyzing Compound Danshen Tablet is explored.

HPLC fingerprint and determination of its main components of Compound Danshen Tablet / 中成药

AIM:The method of HPLC was established for the analysis of fingerprint of Compound Danshen Tablet (Radix et Rhizoma Salvial miltiorrhizal,Radix et Rhizoma Notoginseng,Borneolum Syntheticum) and the determination of its main components. METHODS:The fingerprint chromatograms of liposoluble constituents of Compound Danshen Tablet were set up and its main components of 10 batches were determined. RESULTS:The fingerprint chromatograms were simulated with mean method and the analysis of different batches of Compound Danshen Tablet was made after the analysis of the fingerprint chromatograms from different groups of Compound Danshen Tablet. The similarities which were proved to be higher than 0.99 were calculated with the help of The Similarity Calculation Soft of Fingerprint Chromatography of Traditional Chinese Medicin. CONCLUSION:This method is accurate and provides a scientific basis for controlling the quality of Compound Danshen Tablet.