Compound danshen dripping pills normalize a reprogrammed metabolism of myocardial ischemia rats to interpret its time-dependent efficacy in clinic trials: a metabolomic study.

INTRODUCTION: Clinical trials of Compound danshen dripping pills (CDDP) indicated distinct improvement in patients with chronic stable angina. Daily fluctuation of therapeutic effect agreed with a peak-valley PK profile during a 4-week CDDP regimen, but stabilized after 8-week treatment. OBJECTIVES: This article aims to explore the underlying mechanism for the time-dependent drug efficacy of the up-down fluctuation or stabilization in clinic trials. METHODS: A rat model of myocardial ischemia was established via isoproterenol induction. Metabolomics was employed to analyze the energy-related substances both in circulatory system and myocardium in the myocardial ischemia model. RESULTS: CDDP treatment ameliorated myocardial ischemia, reversed the reprogramming of the metabolism induced by ISO and normalized the level of most myocardial substrates and the genes/enzymes associated with those metabolic changes. After 1- or 2-week treatment, CDDP regulated plasma and myocardial metabolome in an analogous, time-dependent way, and modulated metabolic patterns of ischemic rats that perfectly matched with the fluctuated or stabilized effects observed in clinical trials with 4 or 8-week treatment, respectively. CONCLUSION: Metabolic modulation by CDDP contributes to the fluctuated or stabilized therapeutic outcome, and is a potential therapeutic approach for myocardial ischemia diseases.

Comparison of the effectiveness of whole-brain radiotherapy plus temozolomide versus whole-brain radiotherapy in treating brain metastases based on a systematic review of randomized controlled trials.

Temozolomide (TMZ) combination with whole-brain radiotherapy (WBRT) has been tested by many randomized controlled trials in the treatment of brain metastases (BMs) in China and other countries. We performed an up-to-date meta-analysis to determine (i) the log odds ratios (LORs) of objective response (ORR) and adverse effects (AEs) for all-grade, and (ii) the T value of mean overall survival in patients with BMs treated with WBRT combined with TMZ versus WBRT alone. PubMed, Chinese National Knowledge Infrastructure, and WanFang Data were searched for articles published up to 28 January 2015. Eligible studies were selected according to the PRISMA statement. ORR, AEs, and 95% confidence intervals were calculated using random-effects models. Eighteen studies were included in our analysis. A total of 1028 participants were enrolled. Summary LORs of ORR were 1.0239 (P<0.0001) on comparing WBRT plus TMZ with WBRT ORR (n=17). The overall mean difference of mean overall survival (n=17) between TMZ plus WBRT and WBRT was 2.2505 weeks (P=0.02185). There was a significant difference between WBRT plus TMZ and WBRT alone with a LOR of AEs for all-grade of (i) 0.923 for gastrointestinal toxicity and (ii) 0.7978 for myelosuppression. Sensitivity analysis and subgroup analysis were also performed. The 18 eligible randomized controlled trials demonstrated that the combination of WBRT and TMZ significantly improves the ORR and is statistically insignificant in prolonging the survival of patients with BMs. In addition, an increase in the incidence of gastrointestinal toxicity and myelosuppression was significant for all-grade.

[Identification of liposoluble constituents in Yushu tablets by UPLC-ESI-IT-TOF/MS].

In order to identify the chemical constituents of Yushu tablets comprehensively, we studied the chemical constituents of CHCl3 extract from Yushu tablets by the ultra performance liquid chromatography-electrospray ionization-ion trap-time of flight mass spectrometry (UPLC-ESI-IT-TOF/MS). It showed that there were more than 100 compounds separated, and forty-nine peaks among these were identified on the basis of high resolution mass spectrometry data and literature data reported. Determination of twelve peaks was further confirmed by standard substances. These components assigned to the different plant sources mainly included phenylpropanoids, triterpenoids, quinones and m-trihydroxybenzene compounds. By analyzing the chemical components of CHCl3 extract from compound Chinese medicine Yushu tablets comprehensively, this study provided the foundation for studying chemical components, pharmacodynamic substance and quality control of Yushu tablets.

[Quality assessment of Periplocae Cortex from different habitats by UPLC fingerprint and quantitative analysis].

Ultra performance liquid chromatography (UPLC) was employed for simultaneous determination of three components and fingerprint analysis of Periplocae Cortex with gradient elution of mehtanol and water containing 0.1% phosphoric acid as mobile phase. Three components including chlorogenic acid, 4-methoxysalicylaldehyde and periplocoside were well separated under the analytical condition. Seventeen peaks were selected as the common peaks of 30 batches of Periplocae Cortex. The results showed that there is a significant difference in contents of periplocoside between the samples collected from Henan and Shanxi province. Based on the results of three components quantification and fingerprint analysis, hierarchical clustering analysis ( HCA) and principle component analysis (PCA) were used to further prove the differences between two group samples, and the results indicated that quality of Periplocae Cortex from Shanxi was more stable than that from Henan. The established UPLC fingerprint and quantitative analysis methods could be used efficiently in the quality control of Periplocae Cortex, and this study might contribute to the reasonable clinical application.

Determination and fingerprint analysis of steroidal saponins in roots of Liriope muscari (Decne.) L. H. Bailey by ultra high performance liquid chromatography coupled with ion trap time-of-flight mass spectrometry.

Liriope muscari (Decne.) L. H. Bailey is a well-known traditional Chinese medicine used for treating cough and insomnia. There are few reports on the quality evaluation of this herb partly because the major steroid saponins are not readily identified by UV detectors and are not easily isolated due to the existence of many similar isomers. In this study, a qualitative and quantitative method was developed to analyze the major components in L. muscari (Decne.) L. H. Bailey roots. Sixteen components were deduced and identified primarily by the information obtained from ultra high performance liquid chromatography with ion-trap time-of-flight mass spectrometry. The method demonstrated the desired specificity, linearity, stability, precision, and accuracy for simultaneous determination of 15 constituents (13 steroidal glycosides, 25(R)-ruscogenin, and pentylbenzoate) in 26 samples from different origins. The fingerprint was established, and the evaluation was achieved using similarity analysis and principal component analysis of 15 fingerprint peaks from 26 samples by ultra high performance liquid chromatography. The results from similarity analysis were consistent with those of principal component analysis. All results suggest that the established method could be applied effectively to the determination of multi-ingredients and fingerprint analysis of steroid saponins for quality assessment and control of L. muscari (Decne.) L. H. Bailey.

[Research on UPLC-PDA fingerprint of andrographis paniculata and quantitative determination of 4 major constituents].

Andrographis paniculata from different parts and origins were analyzed by UPLC-PDA fingerprint to provide refererice for related preparation technology. Using the peak of andrographolide as reference, 27 common peaks were identified, and digitized UPLC-PDA fingerprints for 23 batches of andrographis paniculata were established in this research. Principal component analysis (PCA) was carried out after feature extraction. The contents of andrographolide, neoandrographolide, deoxyandrographolide, dehydroandrographolide were determined by external standard method. The Plackett-Burman design combined with pareto chart was used to analyze the factors influencing the robustness of the method. It was found that the medicinal part has a more remarkable influence on the quality of andrographis paniculata than the origin. The contents of the 4 lactones the differ greatly in the different parts of andrographis paniculata, and the pH of the mobile phase is an important factor that influenced the robustness of the method.

[Difference evaluation of three kinds of root of Aconitum carmichaelii in Sichuan based on UPLC analysis of six alkaloids and chemometrics].

An ultra performance liquid chromatography (UPLC) method was established and validated to simultaneously determine the contents of six aconitum alkaloids in mother, daughter and fibrous roots of 19 batches of Aconitum carmichaelii from Sichuan province. The separation of the six alkaloids was achieved on a ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 microm) column at 40 degrees C with a mobile phase consisting of acetonitrile in 30 mmol x L(-1) ammonium acetate buffer solution (adjusted to pH 10.0 with aqueous ammonia) in gradient mode. The data and plots showed that the six aconitum alkaloids have different distributions. Four aconitum alkaloids were almost same in mother and daughter root except benzoylmesaconine and mesaconitine, while the fibrous root differed from the other two roots. The comparisons of significant differences of six aconitum alkaloids between the mother and daughter roots definitely demonstrated that benzoylmesaconine and mesaconitine were the representative components. The 38 detecting samples were classified as two clusters by hierarchical clustering analysis (HCA) and principle component analysis (PCA), the results indicated that the mother root was different from the daughter root on chemical material basis. The study might contribute to the reasonable clinical application of A. carmichaelii.

[Research on ginsenosides composition in wild ginseng leaves in different growing years].

OBJECTIVE: To analyze ginsenosides composition in wild ginseng leaves with different growth years. METHOD: The analysis was performed on Acquity UPLC BEH Shield RP18 (2.1 mm x 100 mm, 1.7 microm) column, the mobile phase was acetonitrile-0.05% formic acid solution in gradient elution mode. The detection wavelength was at 203 nm. The flow rate was 0.4 mL x min(-1) and column temperature was set at 30 degrees C. RESULT: Thirteen ginsenosides were determined by the established UPLC method. In 5-17th growth year ginseng leaf samples cultivated simulating wild conditions, the contents of ginsenosides in the 14th year have the highest content. CONCLUSION: The established method is simple, accurate and reliable, can be used in ginsenosides determination and fingerprint research of Panax crude drug. The result provides reliable data for the accumulation of ginsenosides and sustainable utilization of ginseng resources.

Jinghua Weikang capsule for helicobacter pylori eradication: A systematic review and meta-analysis with trial sequential analysis.

Background: Helicobacter pylori (H. pylori) infection is one of the most common chronic bacterial infections worldwide. The resistance of H. pylori to antibiotics may increase the risk of treatment failure. Complementary and alternative regimens are still needed. This study aimed to critically assess the efficacy and safety of Jinghua Weikang capsule (JWC) for H. pylori eradication. Materials and methods: PubMed, Embase, Web of Science, Cochrane library, China National Knowledge Infrastructure, Wanfang Digital Periodicals, and Chinese Science and Technology Periodicals database were searched from inception to April 2022. Randomized controlled trials (RCTs) comparing a combination of JWC and conventional treatments with conventional treatments alone or combined with a placebo for H. pylori eradication were considered for inclusion. The primary outcome was H. pylori eradication rate. The meta-analysis and trial sequential analysis (TSA) were conducted where possible. Results: A total of 34 studies were included in the statistical analysis. A pooled result showed that JWC with the duration of 2 weeks combined with the triple/quadruple therapy could significantly increase the H. pylori eradication rate compared with the triple/quadruple therapy alone (RR: 1.13, 95% CI: 1.05 to 1.21, p = 0.0008). However, the evidence of benefit was not confirmed by TSA. Another pooled result showed that JWC with the duration of 4 weeks combined with the triple/quadruple therapy could significantly increase the H. pylori eradication rate compared with the triple/quadruple therapy alone (RR: 1.21, 95% CI: 1.15 to 1.27, p < 0.00001). The evidence of benefit was confirmed by TSA. There were no statistically significant differences in the incidence of adverse reactions between the two groups. Conclusion: The present study suggests that JWC with the duration of 4 weeks can significantly improve the H. pylori eradication rate and should be considered as a complementary treatment to conventional regimens for H. pylori eradication. However, more high-quality RCTs are still needed to confirm these findings.

Integrated pharmacokinetics of ginsenosides after intravenous administration of YiQiFuMai powder injection in rats with chronic heart failure by UFLC-MS/MS.

YiQiFuMai powder injection (YQFM), derived from the classical traditional Chinese medicine (TCM) formula Shengmai San, is a modern preparation widely used to combat cardiovascular diseases, chronic heart failure (CHF) for example, in clinical practice in China. Ginsenosides are the major components of YQFM, which are responsible for its therapeutic effect. In this research, we developed a rapid, sensitive and simple method for simultaneous determination of ten ginsenosides from YQFM in CHF rat plasma with ultra-fast liquid chromatography tandem mass spectrometry (UFLC-MS/MS). After solid phase extraction (SPE), chromatography was done on an Acquity UPLC HSS T3 column (1.8µm, 100mm×2.1mm, i.d.) through an 8.0min gradient elution with acetonitrile and 0.1% formic acid in water, while mass spectrometry was performed in the positive ion electrospray ionization (ESI) mode. A good linearity was achieved for each analyte with correlation coefficient (r) >0.9920. The lower limits of quantification (LLOQ) were 1.25ng/mL for ginsenoside Rg1, Rd, Re and Rh1, 2.5ng/mL for ginsenoside Rf, Rg3, Rb2 and Rb3 and 5.0ng/mL for ginsenoside Rb1 and Rc, respectively. All the precision (RSD) data ranged from 1.7-14.5% and the accuracy (RE) data was within ±13.73%. Moreover, the validated method has been applied to investigate the integrated pharmacokinetic profiles of ginsenosides in CHF rats following intravenous administration of YQFM successfully.

Effect of Kaiyu Qingwei Jianji on the morphometry and residual strain distribution of small intestine in experimental diabetic rats.

AIM: To investigate the effect of a Chinese medicine, Kaiyu Qingwei Jianji (KYQWJJ) used for diabetic treatment, on the morphometry and residual strain distribution of the small intestine in streptozotocin (STZ) -induced diabetic rats. Correlation analysis was also performed between the opening angle and residual strain with the blood glucose level. METHODS: Forty-two male Wistar rats weighing 220-240 g were included in this study. Thirty-two STZ-induced diabetic rats were subdivided into four groups (n = 8 in each group), i.e. diabetic control group (DM); high dose of KYQWJJ (T1, 36 g/kg per day); low dose of KYQWJJ (T2, 17 g/kg per day) and Gliclazide (T3, 50 mg/kg per day). Another ten rats were used as non-diabetic control (CON). The medicines were poured directly into stomach lumen by gastric lavage twice daily. The rats of CON and DM groups were only poured the physiological saline. Blood glucose and plasma insulin levels were measured. Experimental period was 35 d. At the end of experiment, three 5-cm long segments were harvested from the duodenum, jejunum and ileum. Three rings of 1-2 mm in length for no-load and zero-stress state tests were cut from the middle of different segments. The morphometric data, such as the circumferential length, the wall thickness and the opening angle were measured from the digitized images of intestinal segments in the no-load state and zero-stress state. The residual strain was computed from the morphometry data. Furthermore, the linear regression analysis was performed between blood glucose level with morphometric and biomechanical data in the different intestinal segments. RESULTS: The blood glucose level of DM group was consistent 4-fold to 5-fold higher than those in CON group during the experiment (16.89+/-1.11 vs 3.44+/-0.15 mmol/L, P < 0.001). The blood glucose level in the T1 (16.89+/-1.11 vs 11.08+/-2.67 mmol/L, P < 0.01) and T3 groups (16.89+/-1.11 vs 13.54+/-1.73 mmol/L, P < 0.05), but not in T2 group (P > 0.05) was significantly lower than those in DM group. The plasma insulin levels of DM, T1, T2 and T3 groups were significantly lower than those in CON group (10.98+/-1.02, 12.52+/-1.42,13.54+/-1.56,10.96+/-0.96 vs 17.84+/-2.34 pmol/L respectively, P < 0.05), but no significantly difference among the groups with exception of CON group. The wet weight/cm and total wall thickness of duodenum, jejunum and ileum in DM group were significantly higher than those in CON group (wet weight (g/cm): duodenum 0.209+/-0.012 vs 0.166+/-0.010, jejunum 0.149+/-0.008 vs 0.121+/-0.004, ileum 0.134+/-0.013 vs 0.112+/-0.007; Wall thickness (mm): duodenum 0.849+/-0.027 vs 0.710+/-0.026, jejunum 0.7259+/-0.034 vs 0.627+/-0.025, ileum 0.532+/-0.023 vs 0.470+/-0.010, all P < 0.05), T1 and T3 treatment could partly restore change of wall thickness, but T2 could not. The opening angle and absolute value of inner and outer residual stain were significantly smaller in duodenal segment (188+/-11 degrees, -0.31+/-0.02 and 0.35+/-0.03 vs 259+/-15 degrees, -0.40+/-0.02 and 0.43+/-0.05) and larger in jejunal (215+/-20 degrees, -0.30+/-0.03 and 0.36+/-0.06 vs 172+/-19 degrees, -0.25+/-0.02 and 0.27+/-0.02) and ileal segments (183+/-20 degrees, -0.28+/-0.01 and 0.34+/-0.05 vs 153+/-14 degrees, -0.23+/-0.03 and 0.29+/-0.04) in DM group than in CON group (P < 0.01). T1 and T3 treatment could partly restore this biomechanical alteration, but strong effect was found in T1 treatment (duodenum 243+/-14 degrees, -0.36+/-0.02 and 0.42+/-0.06, jejunum 180+/-15 degrees, -0.26+/-0.03 and 0.30+/-0.06 and ileum 163+/-17 degrees, -0.23+/-0.03 and 0.30+/-0.05, compared with DM, P < 0.05). The linear association was found between the glucose level with most morphometric and biomechanical data. CONCLUSION: KYQWJJ (high dose) treatment could partly restore the changes of blood glucose level and the remodeling of morphometry and residual strain of small intestine in diabetic rats. The linear regression analysis demonstrated that the effect of KYQWJJ on intestinal opening angle and residual strain is partially through its effect on the blood glucose level.

Qualitative and quantitative analyses of Compound Danshen extract based on 1H NMR method and its application for quality control.

In this study, a new approach using 1H NMR spectroscopy combined with chemometrics method was developed for qualitative and quantitative analyses of extracts of Compound Danshen Dripping Pills (CDDP). For the qualitative analysis, some metabolites presented in Compound Danshen extract (CDE, extraction intermediate of CDDP) were detected, including phenolic acids, saponins, saccharides, organic acids and amino acids, by the proposed 1H NMR method, and metabolites profiles were further analyzed by selected chemometrics algorithms to define the threshold values for product quality evaluation. Moreover, three main phenolic acids (danshensu, salvianolic acid B, and procatechuic aldehyde) in CDE were determined simultaneously, and method validation in terms of linearity, precision, repeatability, accuracy, and stability of the dissolved target compounds in solution was performed. The average recoveries varied between 84.20% and 110.75% while the RSDs were below 6.34% for the three phenolic acids. This 1H NMR method offers an integral view of the extract composition, allows the qualitative and quantitative analysis of CDDP, and has the potential to be a supplementary tool to UPLC/HPLC for quality assessment of Chinese herbal medicines.

Selection of reference standard during method development using the analytical hierarchy process.

Reference standard is critical for ensuring reliable and accurate method performance. One important issue is how to select the ideal one from the alternatives. Unlike the optimization of parameters, the criteria of the reference standard are always immeasurable. The aim of this paper is to recommend a quantitative approach for the selection of reference standard during method development based on the analytical hierarchy process (AHP) as a decision-making tool. Six alternative single reference standards were assessed in quantitative analysis of six phenolic acids from Salvia Miltiorrhiza and its preparations by using ultra-performance liquid chromatography. The AHP model simultaneously considered six criteria related to reference standard characteristics and method performance, containing feasibility to obtain, abundance in samples, chemical stability, accuracy, precision and robustness. The priority of each alternative was calculated using standard AHP analysis method. The results showed that protocatechuic aldehyde is the ideal reference standard, and rosmarinic acid is about 79.8% ability as the second choice. The determination results successfully verified the evaluation ability of this model. The AHP allowed us comprehensive considering the benefits and risks of the alternatives. It was an effective and practical tool for optimization of reference standards during method development.

Novel cytotoxic steroidal glycosides from the roots of Liriope muscari.

The present study was designed to investigate the chemical constituents and bioactivities of the roots of Liriope muscari (Decne.) L.H. Bailey. The compounds were isolated through various chromatography techniques, including silica gel, Sephadex LH-20, and semi-preparative HPLC. The structures were elucidated by infrared (IR), mass spectrometric (MS), 1D- and 2D-NMR analyses in comparison with reference data. In addition, the cytotoxicity of these compounds against human breast cancer MDA-MB-435 cells was evaluated by the MTT assay. Two new steroidal glycosides, 25(R, S)-ruscogenin-1-O-[ß-D-fucopyranosyl (1→2)]-[ß-D-xylopyranosyl(1→3)]ß-D-glucopyranoside (Liriopem I, 1) and 25(R, S)- ruscogenin-1-O-[ß-D-fucopyranosyl (1→2)]-[ß-D-xylopyranosyl(1→4)]-ß-D-fucopyranoside (Liriopem II, 2 and two known compounds LM-S6 (3) and DT-13 (4) were isolated and identified. Liriopem I(1), liriopem II(2) and DT-13 (4) showed remarkable cytotoxicity with IC50 values being (0.58 ± 0.08), (0.05 ± 0.10), and (0.15 ± 0.09) µg·mL(-1), respectively. In summary, compounds 1 and 2 identified in the present study exerted cytotoxicity against breast cancer cells, providing a basis for future development of these compounds as novel anticancer agents.

Simultaneous quantitative determination of six active components in traditional Chinese medicinal preparation Cerebralcare Granule® by RP-HPLC coupled with diode array detection for quality control.

A simple, accurate and reliable method for the simultaneous separation and determination of six active components (protocatechuic acid, chlorogenic acid, caffeic acid, paeoniflorin, ferulic acid and rosmarinic acid) in traditional Chinese medicinal preparation Cerebralcare Granule(®) (CG) was developed using reverse-phase high-performance liquid chromatography coupled with diode array detector detection. The chromatographic separation was performed on a Hypersil GOLD C18 column with aqueous formic acid (0.1%, v/v) and acetonitrile as mobile phase at a flow rate of 0.2 ml/min at 30 °C. Because of the different UV characteristics of these components, change detection wavelength method was used for quantitative analysis. All of the analytes showed good linearity (r > 0.9992). The established method showed good precision and relative standard deviations (%) for intra-day and inter-day variations of 0.15-1.81 and 0.11-1.98%, respectively. The validated method was successfully applied to the simultaneously determination of six active components in CG from different batches.

Identification of metabolites of Radix Paeoniae Alba extract in rat bile, plasma and urine by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry.

Ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF/MS) was developed to identify the absorbed parent components and metabolites in rat bile, plasma and urine after oral administration of Radix Paeoniae Alba extract (RPAE). A total of 65 compounds were detected in rat bile, plasma and urine samples, including 11 parent compounds and 54 metabolites. The results indicated that glucuronidation, hydroxylation and methylation were the major metabolic pathways of the components of RPAE. Furthermore, the results of this work demonstrated that UPLC-Q-TOF/MS combined with MetaboLynx™ software and mass defect filtering (MDF) could provide unique high throughput capabilities for drug metabolism study, with excellent MS mass accuracy and enhanced MSE data acquisition. With the MSE technique, both precursor and fragment mass spectra can be simultaneously acquired by alternating between high and low collision energy during a single chromatographic run.

Chinese herbal medicine for obesity: a randomized, double-blinded, multicenter, prospective trial.

Obesity is a serious medical problem worldwide. As a holistic therapy, traditional Chinese medicine (TCM) may have a potential in obesity management. In this controlled trial, we evaluated the safety and effectiveness of xin-ju-xiao-gao-fang (XJXGF), a TCM herbal formulation, in 140 obese subjects over a 24-week period. The XJXGF formula mainly consists of rhubarb, coptis, semen cassia, and citrus aurantium. Subjects with body mass index (BMI) 28-40 kg/m(2) were recruited at 5 centers in China. We assessed the changes in subjects' body weight, its related parameters, and the reduction of insulin resistance (IR) after administration of XJXGF formula or low-dose XJXGF (10% of the XJXGF formula, as control). After 24-week treatment, among participants in the XJXGF formula group and low-dose XJXGF group, the mean ± SE changes in the body weight were -3.58 ± 0.48 and -1.91 ± 0.38 kg, respectively (p < 0.01). The changes in the IR-index of two groups were -2.65 ± 1.04 and -1.58 ± 1.3, respectively (p < 0 .05). There were no serious adverse events reported during the 24-week trial. Participants reported 7 minor adverse events, 4 in the XJXGF formula group and 3 in the low-dose XJXGF group (p = 0.578). Future studies are needed to investigate the clinical utility of this TCM formulation in the treatment of obese subjects.

Salvianolic acids T and U: a pair of atropisomeric trimeric caffeic acids derivatives from root of Salvia miltiorrhiza.

Two new trimeric caffeic acids, named salvianolic acids T and U (1 and 2), were isolated from the underground part of Salvia miltiorrhiza. Their structures, consisting of three caffeic acid units, were determined based on extensive 1D- and 2D-spectroscopic analyses and electronic circular dichroism (ECD) calculations.

Simultaneous determination of five phenolic components and paeoniflorin in rat plasma by liquid chromatography-tandem mass spectrometry and pharmacokinetic study after oral administration of Cerebralcare granule(®).

A simple, sensitive and selective high-performance liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) method was developed for simultaneous determination and pharmacokinetic study of six active components, protocatechuic acid, chlorogenic acid, caffeic acid, ferulic acid rosmarinic acid and paeoniflorin in rat plasma after oral administration of Cerebralcare granule(®) for the first time. The method involves a simple liquid-liquid extraction with ethyl acetate. The separation was performed on a Luna C18 column (2.0×100mm i.d., 3.0µm, particle, Phenomenex, USA) with gradient elution using a mobile phase composed of acetonitrile and water (containing 0.1% formic acid) at a flow rate of 0.2ml/min. Electrospray ionization (ESI) in negative ion mode and selective reaction monitoring (SRM) was used for the quantification of six active components and internal standard (IS, Chloroamphenicol). The method was linear for all analytes over investigated range with all correlation coefficients greater than 0.9914. The lower limits of quantification (LLOQ) were 1.0ng/ml for protocatechuic acid, 1.0ng/ml for chlorogenic acid, 1.0ng/ml for caffeic acid, 5.0ng/ml for ferulic acid, 1.5ng/ml for rosmarinic acid and 6.0ng/ml for paeoniflorin, respectively. The intra- and inter-day precisions (R.S.D.%) were less than 6.60% and 11.68%, and accuracy (RE %) between -3.26% and 1.13% (n=6). The developed method was applied for the first time to the pharmacokinetic study of protocatechuic acid, chlorogenic acid, caffeic acid, ferulic acid, rosmarinic acid and paeoniflorin in rat plasma after oral administration of Cerebralcare granule(®).

Determination of ginsenoside Rc in rat plasma by LC-MS/MS and its application to a pharmacokinetic study.

Ginsenoside Rc (GRc) is a potential pharmacologically active ingredient isolated from ginseng (Panax ginseng C.A. Meyer, Araliaceae). A simple, rapid and sensitive method for determination of GRc in rat plasma was developed based on liquid chromatography-tandem mass spectrometry (LC-MS/MS). The analyte and internal standard (IS), ginsenoside Rb(1) (GRb(1)), were extracted from plasma with n-butanol and chromatographied on a C(18) column eluted with a mobile phase of methanol and water containing 0.1% formic acid. The detection was performed by positive ion electrospray ionization in selective reaction monitoring mode (SRM), monitoring the transitions m/z 1101.6→789.3 and m/z 1131.7→364.7 for GRc and IS, respectively. The assay was linear over the concentration range of 5-5000 ng/mL with a limit of quantitation (LOQ) of 5 ng/mL. The accuracy was between 86.7% and 114.9%, and the precision was less than 9.7%. This method was successfully applied to investigate the pharmacokinetic study of GRc in rats after intravenous (2mg/kg) and oral (20mg/kg) administration, and the result showed that the ginsenoside was poorly absorbed with an absolute bioavailability being approximately 0.17%.