Danshen Preparations Research Articles

Clinical efficacy of Danshen preparation in the treatment of vascular cognitive impairment: A systematic review and meta-analysis.

Danshen preparations are widely used in the treatment of ischaemic cerebrovascular disease. However, the clinical efficacy of such preparations remains unclear. Consequently, Danshen preparations are used to a lesser extent in vascular cognitive impairment (VCI). In this study, we aimed to systematically assess the clinical efficacy and safety of Danshen preparations in VCI. To this end, we examined and performed a meta-analysis (MA) of the evidence available from randomised controlled trials (RCTs) of Danshen preparations conducted in patients with VCI. We queried the following sources and collected all articles reporting on RCTs of Danshen preparations published prior to December 2021: PubMed, China National Knowledge Infrastructure (CNKI), Wanfang Data, Chongqing VIP Database (CQVIP), and China Biology Medicine (CBM) disc databases. The assessment of treatments that were included in references were performed by RevMan 5.2 software based on guidelines from Cochrane Handbook for Systematic Reviews of Interventions. We included a total of 12 RCTs that included data on clinical therapeutic effects. The evaluation criteria included the following: National Institutes of Health Stroke Scale (NIHSS), modified Rankin Scale (mRS), Barthel Index (BI), Mini-Mental State Assessment (MMSE), Montreal Cognitive Assessment (MOCA), Activities of Daily Living Scale (ADL), treatment effect index, and incidence of adverse reaction index. In the included studies, the observation groups included 656 cases and the control groups included 660 cases. The results of the MA were as follows: the mean difference (MD) value after combining the effect size for NIHSS was -2.91, with a 95% confidence interval (CI) of -4.22 to -1.59; the combined effect quantity hypothesis test revealed that Z = 4.33 (p < 0.00001), indicating that the score pertaining to the degree of neurological deficit (NIHSS) in the observation group after treatment was significantly lower than that in the control group. This result reveals that treatment with a Danshen preparation can reduce neurological deficit in VCI patients. The MD value after combining the effect size for mRS was -0.73, with a 95% CI of -0.85 to -0.61; the result of the combined effect quantity hypothesis test revealed that Z = 12.29 (p < 0.00001). These results indicate that the degree of disability was significantly reduced after treatment in the observation group. The MD value after combining the effect size for MMSE was 2.09, with a 95% CI of 0.33-3.84; the result of the combined effect quantity hypothesis test showed that Z = 2.33 (p = 0.02). These results indicate that the cognitive function score (MMSE) of the observation group after treatment was significantly higher than that of the control group and suggests that the cognitive function of VCI patients was improved after treatment with Danshen preparations. The MD value after combining the effect size for ADL was 8.79, with a 95% CI of 3.52 to 14.06; the result of the combined effect quantity hypothesis test indicated that Z = 3.27 (p = 0.001). These results showed that the life ability (ADL scale) scores of patients in the observation group after treatment were significantly higher than those in the control group, and reveals that after treatment with Danshen preparations, patients exhibited significant improvement in life ability. The MD value after combining the effect size for high-sensitivity C-reactive protein (hs-CRP) was -3.21, with a 95% CI of -4.21 to -2.22; the result of the combined effect quantity hypothesis test showed that Z = 6.31 (p < 0.00001), indicating that the hs-CRP level in the observation group was significantly decreased after treatment. The MD value after combining the effect size for interleukin (IL)-6 was -2.95, with a 95% CI of -3.86 to -2.04; the result of the combined effect quantity hypothesis test showed that Z = 6.36 (p < 0.00001). These results showed that the IL-6 level in the observation group was significantly decreased after treatment. The existing clinical evidence shows that Danshen preparations exert significant therapeutic effects on VCI patients and can ameliorate inflammatory conditions in these patients. In addition, Danshen preparations are relatively safe.

In vitro inhibitory effects of components from Salvia miltiorrhiza on catalytic activity of three human Arachidonic acid ω-hydroxylases

CYP4 enzymes are involved in the metabolism of xenobiotics and endogenous molecules. 20-Hydroxyeicosatetraenoic acid (20-HETE), the arachidonic acid (AA) ω-hydroxylation metabolite catalyzed by CYP4A/4F enzymes, is implicated in various biological functions. The goal of this investigation is to examine the inhibitory effects of components from Salvia miltiorrhiza(Danshen) on AA ω-hydroxylation using recombinant CYP4A11, CYP4F2, CYP4F3B, and microsomal systems. Tanshinone IIA had noncompetitive inhibition on CYP4F3B (Ki = 4.98 μM). Cryptotanshinone (Ki = 6.87 μM) and tanshinone I (Ki = 0.42 μM) had mixed-type inhibition on CYP4A11. Dihydrotanshinone I had mixed-type inhibition on CYP4A11 (Ki = 0.09 μM), and noncompetitive inhibition on CYP4F2 (Ki = 4.25 μM) and CYP4F3B (Ki = 3.08 μM). Salvianolic acid A had competitive inhibition on CYP4A11 (Ki = 19.37 μM), and noncompetitive inhibition on CYP4F2 (Ki = 15.28 μM) and CYP4F3B (Ki = 6.45 μM). Salvianolic acid C had noncompetitive inhibition on CYP4F2 (Ki = 5.70 μM) and CYP4F3B (Ki = 18.64 μM). In human kidney, human liver or rat heart microsomes, 20-HETE formation was significantly inhibited (P < 0.05) by dihydrotanshinone I (5 and 20 μM) and salvianolic acid A (20 and 50 μM). Given that low plasma concentrations of Danshen components after oral administration, Danshen preparations may not play pharmacological roles by inhibiting AA ω-hydroxylases; however, as Danshen components may reach high concentration in human intestine, drugs that have an important pre-systemic metabolism by these CYP4A/4F enzymes should avoid being co-administered with Danshen preparations.

Study on the Quality Evaluation of Compound Danshen Preparations Based on the xCELLigence Real-Time Cell-Based Assay and Pharmacodynamic Authentication.

Objective: To perform a preliminary study on the quality evaluation of compound Danshen preparations based on the xCELLigence Real-Time Cell-based Assay (RTCA) system and make a pharmacodynamics verification. Methods: The compound Danshen was discussed as a methodological example, and the bioactivity of the compound Danshen preparations were evaluated by real-time cell electronic analysis technology. Meanwhile, an in vivo experiment on an acute blood stasis rat model was performed in order to verify this novel evaluation through the curative effect of dissipating blood stasis. Results: We determined the cell index (CI) and IC50 of the compound Danshen preparations and produced time/dose-dependent cell response profiles (TCRPs). The quality of the three kinds of compound Danshen preparations was evaluated through the RTCA data. The trend of CI and TCRPs reflected the effect of drugs on the cell (promoting or inhibiting), and it was verified that the results correlated with the biological activity of the drugs using a pharmacodynamics experiment. Conclusion: The RTCA system can be used to evaluate the quality of compound Danshen Preparations, and it can provide a new idea and new method for quantitatively characterizing the biological activity of traditional Chinese medicines (TCMs).

Inhibitory Effects of Danshen components on CYP2C8 and CYP2J2

The use of Chinese herbal medicines and natural products has become increasingly popular in both China and Western societies as an alternative medicine for the treatment of diseases or as a health supplement. Danshen, the dried root of Salvia miltiorrhiza (Fam.Labiatae), which is rich in phenolic acids and tanshinones, is a widely used herbal medicine for the treatment of cardio-cerebrovascular diseases. The goal of this study was to examine the inhibitory effects of fifteen components derived from Danshen on CYP2C8 and CYP2J2, which are expressed both in human liver and cardiovascular systems. Recombinant CYP2C8 and CYP2J2 were used, and the mechanism, kinetics, and type of inhibition were determined. Taxol 6-hydroxylation and astemizole O-desmethyastemizole were determined as probe activities for CYP2C8 and CYP2J2, respectively. Metabolites formations were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results demonstrated that salvianolic acid A was a competitive inhibitor of CYP2C8 (Ki = 2.5 μM) and mixed-type inhibitor of CYP2J2 (Ki = 7.44 μM). Salvianolic acid C had moderate noncompetitive and mixed-type inhibitions on CYP2C8 (Ki = 4.82 μM) and CYP2J2 (Ki = 5.75 μM), respectively. Tanshinone IIA was a moderate competitive inhibitor of CYP2C8 (Ki = 1.18 μM). Dihydrotanshinone I had moderate noncompetitive inhibition on CYP2J2 (Ki = 6.59 μM), but mechanism-based inhibition on CYP2C8 (KI = 0.43 μM, kinact = 0.097 min−1). Tanshinone I was a moderate competitive inhibitor of CYP2C8 (Ki = 4.20 μM). These findings suggested that Danshen preparations appear not likely to pose a significant risk of drug interactions mediated by CYP2C8 after oral administration; but their inhibitory effects on intestinal CYP2J2 mediated drug metabolism should not be neglected when they are given orally in combination with other drugs. Additionally, this study provided novel insights into the underling pharmacological mechanisms of Danshen components from the perspective of CYP2C8 and CYP2J2 inhibition.

Different network pharmacology mechanisms of Danshen-based Fangjis in the treatment of stable angina.

Danshen (Salvia miltiorrhiza) preparations such as Danhong injection, Danshen injection, Salvianolate injection, compound Danshen injection and Sodium Tanshinone IIA Sulfonate (STS) injection are widely used in China to treat stable angina (angina pectoris) caused by coronary heart disease. In this study we compared the network pharmacological mechanisms of the 5 Danshen preparations. Following a literature search performed in PubMed, EMBASE, Cochrane Library, China National Knowledge Infrastructure (CNKI) database, China Biology Medicine (CBM) database, China Conference Paper Database, Wanfang Database, VIP Database and Conference Proceedings Citation Index (through January 2015), 444 randomized controlled trial publications detailing the use of the 5 Danshen-based injections for treating stable angina were identified, and their combined data were analyzed using a network meta-analysis. All of the 5 Danshen-based preparations were effective in treating stable angina with clinical improvement rates of 72.4%-91.6% and electrocardiogram (ECG) improvement rates of 54.5%-71.6%. According to both clinical improvement and ECG improvement, the 5 Danshen-based preparations were ranked as follows: Danhong injection > Salvianolate injection > STS injection > compound Danshen injection > Danshen injection. There were no significant differences among the safety profiles of the 5 Danshen preparations. The meta-analysis results were further examined using a network pharmacology approach and functional enrichment analysis, which revealed that Danshen and Danhong injections affected 4 and 15 signaling pathways, respectively, and that the 4 signaling pathways affected by Danshen were a subset of those influenced by Danhong. Therefore, Danhong injection affected some unique signaling pathways that might regulate lipoprotein metabolism, oxidation, and inflammation, and protect vascular endothelia, reflecting the multi-component and multi-target characteristics of this traditional formula and its strengths in treating complex diseases.

Effective dose of the combination of salvianolic acids and tanshinones against inflammatory injury in human umbilical vein endothelial cells injured by thrombin.

To investigate the pharmacological effective doses of Chinese Danshen components, (salvianolic acids and tanshinones) combinations on vascular endothelia cells against inflammatory injury. Thrombin (50 U/mL) was incubated in human umbilical vein endothelia cells (HUVECs) with 50 and 100 μg/L (1:0, 0:1, 3:1, 2:1, 1:1, and 1:2) salvianolic acids and tanshinones or without the components of Danshen extract. Cell viability was confirmed in (4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium (MTT) assay and cell death was confirmed in lactate dehydrogenase (LDH) released into the cell culture supernatant. Six hours treatment of HUVECs with 50 or 100 μg/L salvianolic acids and tanshinones combinations at ratios 1:0, 0:1, 3:1, 2:1, 1:1, and 1:2 significantly (P<0.05 or P<0.01) improved cell proliferation in vitro compared with model group when assayed in MTT, whereas 100 μg/L groups displayed trend toward upshift in potency compared with 50 μg/L groups (2:1, 1:1, and 1:2), respectively, in addition to inhibition of cell death confirmed in reduced LDH released into the culture medium. These findings provided some useful information for safe and effective use of Danshen preparations in clinical practice. We made observations regarding the protective effect of Danshen components combinations in vascular endothelial against inflammatory injury caused by thrombin activation of endothelial cell.

Establishment and validation of quantitative analysis of multi-components by single-marker for quality assessment of compound danshen preparations

Compound danshen preparations (CDPs) are used clinically for the treatment of cardiovascular and cerebrovascular diseases. By using the quantitative analysis of multi-components by single-marker (QAMS) method, sixteen compounds (danshensu, protocatechuic acid, protocatechuicaldehyde, caffeic acid, rosmarinic acid, lithospermic acid, notoginsenoside R1, salvianolic acid B, ginsenoside Rg1, ginsenoside Re, salvianolic acid A, salvianolic acid C, ginsenoside Rb1, ginsenoside Rd, cryptotanshinone, and tanshinone IIA were quantified on an ACQUITY ultraperformance liquid chromatography (UPLC) HSS T3 column (2.1 × 100 mm, 1.8 μm) with the mobile phase consisting of 0.1% formic acid aqueous solution (A) and acetonitrile (B) using a gradient elution at the flow rate of 0.30 mL/min in 30 min at 30°C, which was also validated by UPLC-diode array detection (DAD) and UPLC-electrospray ionization multistage/mass spectrometry (ESI-MS/MS) for assuring the feasibility and accuracy. Tested by robustness experiment under slightly changeable conditions, the stability of relative correction factor (RCF) proved to be stable, with RSDs below 5.69%, except for notoginsenoside R1 with relative standard deviation (RSD) 7.83%. This reliable and convenient QAMS method resolved the problem of standard substance insufficiency and improved the quality assessment of preparations consisting of complex compounds with different chemical structures, such as CDPs.

Herb-drug interactions with Danshen (Salvia miltiorrhiza): a review on the role of cytochrome P450 enzymes

Danshen, the dried root and rhizome of Salvia miltiorrhiza Bunge, is a widely used medicinal plant for the treatment of cardiovascular diseases in China and a complementary medicine in the West. Danshen is indexed in the 2010 Chinese Pharmacopoeia, with more than 35 formulations and concoctions containing Danshen water-extracts, ethanolic extracts or their combination, which are rich in phenolic acids and different levels of tanshinones. There are rare reports on the adverse effects of Danshen preparations. It is, however, well-known that Danshen leads the anticoagulation failure of warfarin. The Danshen-warfarin interaction may be mediated via both pharmacodynamic and pharmacokinetic mechanisms. This review does not summarize recent progress, but the effects of Danshen and its active ingredients on the interactions of cytochrome P450 (CYP450) and drug transporters, as well as the analysis of ingredients, and the metabolism and pharmacokinetics that are related to these interactions. Tanshinones play significant roles in the inhibition and induction of several CYP450 isozymes. It can be concluded that precautions should be taken when using Danshen preparations rich in tanshinones for CYP-related herb-drug interactions.

Major tanshinones of Danshen (Salvia miltiorrhiza) exhibit different modes of inhibition on human CYP1A2, CYP2C9, CYP2E1 and CYP3A4 activities in vitro

This study investigated the effects of tanshinones on human CYP1A2 (phenacetin O-deethylase), CYP2C9 (tolbutamide 4-hydroxylase), CYP2E1 (chlorzoxazone 6-hydroxylase) and CYP3A4 (testosterone 6beta-hydroxylase) activities in vitro using pooled human liver microsomes and specific human CYP isoforms. Tanshinone I, tanshinone IIA, and cryptotanshinone were potent competitive CYP1A2 inhibitors (K(i)=1.5-2.5 microM); medium competitive inhibitors of CYP2C9 (K(i)=22-62 microM); medium competitive inhibitors of CYP2E1 (K(i)=3.67 microM) for tanshinone I and 10.8 microM for crytotanshinone; but weak competitive inhibitors of CYP3A4 (K(i)=86-220 microM). Dihydrotanshinone was a competitive inhibitor of human CYP1A2 (K(i)=0.53 microM) and CYP2C9 (K(i)=1.92 microM), a noncompetitive inhibitor of CYP3A4 (K(i)=2.11 microM) but an uncompetitive CYP2E1 inhibitor. In conclusion, these results showed that tanshinones inhibited the metabolism of various CYP probe substrates in human liver microsomes and specific human CYP isoforms in vitro. Given that CYP1A2, 2C9, 2E1 and 3A4 are responsible for the metabolism and disposition of a large number of drugs currently used, the potential herb-drug interactions of Danshen preparations containing the major tanshinones with drugs which are substrates of these CYPs may be important.

Simultaneous determination of danshensu, rosmarinic acid, cryptotanshinone, tanshinone IIA, tanshinone I and dihydrotanshinone I by liquid chromatographic–mass spectrometry and the application to pharmacokinetics in rats

A sensitive and specific liquid chromatography–tandem mass spectrometry method (LC–MS) was developed and validated for the separation and simultaneous determination of danshensu, rosmarinic acid and tanshinone compounds including cryptotanshinone, tanshinone I, dihydrotanshinone I and tanshinone IIA in rat plasma. Chromatographic separation of the analytes was successfully achieved on a C 18 column using a mobile phase composed of acetonitrile–water containing 0.5% glacial acetic acid. This method demonstrated good linearity and did not have endogenous material interfering with the active compounds and I.S. peaks. The limit of quantification of danshensu, rosmarinic acid, cryptotanshinone, dihydrotanshinone I, tanshinone I and tanshinone IIA were 5, 0.75, 0.1, 0.1, 1 and 0.5 ng/mL. The average extraction recoveries of these analytes from rat plasma were all over 60%. The precisions determined from five days were all within 10%. This method has been successfully applied in the simultaneous quantification and the pharmacokinetic studies of these six compounds in animals which were orally administered with danshen preparations.

CHINESE MEDICINE (CM) matters

CHINESE MEDICINE (CM) matters

Determination of Phenolic Acids in Danshen Preparations by LC with Chemiluminescence Detection

A simple and sensitive method has been developed for the simultaneous determination of eight phenolic acids in Danshen preparations based on liquid chromatography with chemiluminescence detection. Chemiluminescence parameters including flow rate, buffer pH, the concentration of luminescent and reactive solutions, were optimized. The analytical performance of the optimized luminol-H2O2 detection was compared with those of luminol-pyrogallol and UV detection. Under the optimized conditions, the method was validated with respect to linearity, precision, limits of detection and quantification. The method offers an attractive alternative to be used to evaluate the quality of Danshen preparations.

Application of liquid chromatography–electrospray ionization time‐of‐flight mass spectrometry for analysis and quality control of compound Danshen preparations

A liquid chromatography coupled with electrospray ionization time-of-flight mass spectrometry (LC-ESI-TOF-MS) method has been developed to evaluate the quality of three formulas of compound Danshen preparations (CDPs), through a simultaneous determination of 22 marker constituents (nine major phenolic acids, eight major saponins and five major diterpenoids). Optimum separations were obtained with a Zorbax C(18) column, using a gradient elution with 0.1% aqueous formic acid and acetonitrile. Limits of detection and quantification were in ranges of 1.58-10.10 and 4.85-28.56 ng/mL. All calibration curves showed good linear regression (r(2 ) > 0.9900) within the test range, and the recoveries were between 78.4 and 103.1% for all analytes. The assay was successfully utilized to analyze the 22 marker components in 26 samples. The overall results demonstrated that this method is sensitive, accurate and reliable for the quality control of CDPs.

Rapid and sensitive analysis of multiple bioactive constituents in Compound Danshen preparations using LC‐ESI‐TOF‐MS

By fully optimizing the separation and analytical conditions, a rapid-resolution LC (RRLC, 1.8 microm particle size) coupled with TOF-MS method was developed and validated for multiple compounds analysis of traditional Chinese medicinal compound preparations (TCMCPs). As a typical example, determination of Compound Danshen preparations (CDPs, mainly comprising Salvia miltiorrhiza and Panax notoginseng), was improved on three groups of bioactive constituents (phenolic acids, diterpenoids, and saponins). LODs down to 1.2 pg, permitting very good separation of 20 analytes and 3 internal standards (ISs) in 17 min, represented an approximate five-fold reduction of time compared to conventional HPLC. Unequivocal identification of target compounds in CDPs was based on accurate mass measurement of ions, and comparison of retention performance with available standards. Quantitation was carried out using the narrow window extracted ion chromatograms (XICs) of each compound with an interference-free baseline (using a +/-0.02 Da window) yielding good linearity of response (r(2) >0.9940), and excellent precision of retention time (t(R)) both interday (RSD, 0.01-0.19%) and intraday (RSD, 0.02-1.08%). The acceptable recoveries obtained were in the range of 77.69-113.8%. The results demonstrated the robustness and applicability of RRLC-TOF-MS for multicomponents analysis in TCMCPs with diverse chemical compositions and properties.

Danshen (Chinese medicinal herb) preparations for acute myocardial infarction.

Acute myocardial infarction (AMI) is the most important cause of morbidity from ischaemic heart disease, and is among the leading causes of death in the western world. Danshen, a Chinese herbal medicine, is widely used in China for treatment of several diseases, including AMI. To assess the effects (both benefits and harms) of danshen preparations for AMI. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) on The Cochrane Library (issue 4, 2006), MEDLINE (1966-2006), EMBASE (1980-2006), and the Chinese Biomedical Database (CBM) (1982-2006). We also handsearched 75 Chinese medical journals. Randomised controlled trials (RCTs) lasting at least 7 days were sought. Since it seemed evident that few RCTs were available, we also considered other controlled studies. Eligibility and trial quality were assessed by three reviewers. Six studies comprised of 2368 participants were included. Only one trial was judged to be a genuine RCT and showed no statistically significant difference in reduction of total mortality (Peto OR 0.55, 95% CI 0.23 to 1.32), but a quasi-RCT reported a reduced total mortality (Peto OR 0.42, 95% CI 0.23 to 0.77). Pooling these trials yielded an approximate halving of mortality in those patients treated with danshen preparations plus usual care compared with usual care alone (Peto OR 0.46, 95% CI 0.28 to 0.75). The evidence to support use of danshen preparations is too weak to make any judgement about its effects. Evidence from RCTs is insufficient and of low quality. The safety of danshen preparations is unproven, although some adverse events have been reported. More evidence from high quality trials is needed to support the clinical use of danshen preparations.

Inhibitory Effects of Seven Components of Danshen Extract on Catalytic Activity of Cytochrome P450 Enzyme in Human Liver Microsomes

The potential for herb-drug interactions has recently received greater attention worldwide, considering the fact that the use of herbal products becomes more and more widespread. The goal of this work was to examine the potential for the metabolism-based drug interaction arising from seven active components (danshensu, protocatechuic aldehyde, protocatechuic acid, salvianolic acid B, tanshinone I, tanshinone IIA, and cryptotanshinone) of danshen extract. Probe substrates of cytochrome P450 enzymes were incubated in human liver microsomes (HLMs) with or without each component of danshen extract. IC(50) and K(i) values were estimated, and the types of inhibition were determined. Among the seven components of danshen extract, tanshinone I, tanshinone IIA, and cryptotanshinone were potent competitive inhibitors of CYP1A2 (K(i) = 0.48, 1.0, and 0.45 microM, respectively); danshensu was a competitive inhibitor of CYP2C9 (K(i) = 35 microM), and cryptotanshinone was a moderate mixed-type inhibitor of CYP2C9 (K(i) = 8 microM); cryptotanshinone inhibited weakly and in mixed mode against CYP2D6 activity (K(i) = 68 microM), and tanshinone I was a weak inhibitor of CYP2D6 (IC(50) = 120 microM); and protocatechuic aldehyde was a weak inhibitor of CYP3A4 (IC(50) = 130 and 160 microM for midazolam and testosterone, respectively). These findings provided some useful information for safe and effective use of danshen preparations in clinical practice. Our data indicated that it was necessary to study the in vivo interactions between drugs and pharmaceuticals with danshen extract.

Chromatographic fingerprinting analysis of Danshen root (Salvia miltiorrhiza Radix et Rhizoma) and its preparations using high performance liquid chromatography with diode array detection and electrospray mass spectrometry (HPLC‐DAD‐ESI/MS)

Danshen root (Salvia miltiorrhiza Radix et Rhizoma) is a representative Chinese herb containing multiple components contributing to its polyvalent bioactivities. Advanced analysis approaches are needed to obtain a comprehensive picture of the targeting constituents in complete matrix. In this study, a chromatographic fingerprinting method to monitor simultaneously the hydrophilic and lipophilic constituents was developed for the quality evaluation of Danshen root and its preparations. Ten hydrophilic and nine lipophilic components were identified using HPLC-diode array detection-electrospray-MS (HPLC-DAD-ESI/MS) by comparing with the available references and reported data. Using the established method, 13 Danshen root samples collected from different sources, and 21 batches of Danshen preparations including tablets, injections, capsules, and dropping pills produced by different manufacturers were analyzed and their chromatographic fingerprints (CFP) were constructed. The results showed that the products of Danshen roots such as the tablets and capsules contained both the hydrophilic and lipophilic components, while the injections and dropping pills contained mainly the hydrophilic components. Principal components analysis (PCA) was applied for the statistical analysis of the fingerprinting data of crude herb and its preparations. The established CFPs demonstrate the representative chemical profiling of the existing components and can be applied to the authentication and quality assessment of Danshen roots and other Danshen containing formulated preparations.

Improved quality control method for Fufang Danshen preparations through simultaneous determination of phenolic acids, saponins and diterpenoid quinones by HPLC coupled with diode array and evaporative light scattering detectors

A herb-combined prescription, mainly derived from roots of Salvia miltiorrhiza and Panax notoginseng, has been widely used for improving coronary or cerebral circulation in China as well as in Western countries. Multiple commercially available preparations, known as Fufang Danshen preparations (FDPs), produced by various manufacturers with the raw materials from different sources, pose a serious challenge to the quality control of this herb medicine. Previous pharmacological studies identify three types of bioactive components correlated with the clinical effect of those herb preparations. Those mainly include four phenolic acids, four saponins and four diterpenoid quinones. In this report, by using high performance liquid chromatography (HPLC) coupled with diode array and evaporative light scattering detectors (DAD–ELSD), we developed an improved quality control method for those herb medicines. A simultaneous separation and quantification of the 12 components was performed on a C 18 column, in which the mobile phase consisted of (A) 0.1% aqueous formic acid and (B) acetonitrile using a gradient elution. The optimum detection wavelength was set at 281 nm, the drift tube temperature of ELSD was set at 113 °C, the nitrogen flow rate at 3.1 L/min, and the gain = 4. All calibration curves showed good linear regression ( r 2 > 0.9927) within test ranges. The method developed showed good precision and accuracy with overall intra- and inter-day variations of 0.64–4.79% and 0.69–4.96%, respectively, and the overall recoveries of 93.50–107.69% for the 12 compounds analyzed. This method was successfully applied to quantify the twelve components in ten commercial samples from three formulas by seven independent manufacturers. This readily available, low-cost and reliable HPLC–DAD–ELSD method improved the quality control of traditional Chinese medicinal preparations consisting of complex compounds with different structures such as FDPs.

New Enzyme-Linked Chemiluminescent Immunosorbent Digoxin Assay Is Free From Interference of Chinese Medicine DanShen

DanShen is a traditional Chinese medicine indicated for cardiovascular diseases. The potential interference of DanShen with serum digoxin measurement was investigated using a new enzyme-linked chemiluminescent immunosorbent (ECLIA) digoxin assay. Aliquots of drug-free serum were supplemented with ethyl acetate extract of DanShen (4 different brands studied), and apparent digoxin concentrations were measured by the ECLIA as well as fluorescence polarization immunoassay (FPIA) and a turbidimetric assay for comparison. Mice were also fed 4 DanShen preparations and apparent digoxin concentrations were subsequently measured. In another experiment, serum pools containing digoxin were further supplemented with DanShen extracts and digoxin concentrations were measured again by all 3 assays. No apparent digoxin concentration was observed when aliquots of drug-free serum pools were supplemented with DanShen and digoxin concentrations were measured by the ECLIA or the turbidimetric assay. In contrast, significant apparent digoxin concentrations were observed using FPIA, and the highest apparent digoxin concentration was observed with brand 4 of DanShen extract. Similarly, when mice were fed with this herb, significant apparent digoxin concentrations were also observed using FPIA, but neither ECLIA nor turbidimetric assay showed any apparent digoxin concentration. When aliquots of digoxin pool were further supplemented with various DanShen extract, the apparent digoxin concentrations were significantly increased when FPIA was used. In contrast, digoxin concentrations in the presence of DanShen extract compared well with the digoxin concentration of the original pool when ECLIA or turbidimetric assay was used. We conclude that DanShen does not interfere with serum digoxin measurement using a more recently released ECLIA digoxin assay.