Comprehensive Evaluation of the Quality of Tripterygium Glycosides Tablets Based on Multi-Component Quantification Combined with an In Vitro Biological Assay.

Tripterygium glycosides tablets (TGTs) are widely used in clinical practice to treat rheumatoid arthritis and other autoimmune diseases, with significant beneficial effects but also high toxicity, necessitating rigorous quality evaluation and control. In current study, a rapid resolution liquid chromatography tandem electrospray ionization triple quadrupole mass spectrometry (RRLC-ESI-MS/MS) method was developed and validated for the quantitative analysis of 14 components of ten batches of TGTs produced by different manufacturers, including four diterpenoids, three triterpenoids, and seven sesquiterpene alkaloids. Meanwhile, the NO inhibition effects of these TGTs were evaluated in LPS-induced RAW264.7 cells for their downstream anti-inflammatory activities, as well as their cytotoxicity. The results indicate that the TGTs from different manufacturers showed poor quality consistency, as evidenced by large variations in chemical profiles and biological effects, which may increase the risks associated with clinical use. To improve the quality status of TGTs, it is crucial to identify indicator components whose characterization can accurately reflect the efficacy and toxicity of TGTs from which they were derived. Our study reveals that triptolide, triptoquinone B, celastrol, and demethylzelaysteral considerably contributed to the anti-inflammatory activity and/or cytotoxicity of TGTs, implying that they should be further investigated as candidate indicator components for TGT quality control.

Plasma Ceramides in Relation to Coronary Plaque Characterization Determined by Optical Coherence Tomography.

Plasma ceramides (Cer), a subset of bioactive lipids, have mechanistic links to development of atherosclerosis and are related to major adverse cardiovascular events (MACEs). Previous researches have demonstrated vulnerable plaques contribute to acute cardiovascular events and poor prognosis. This study aimed to explore the associations between Cer and culprit plaque characterizations evaluated by optical coherence tomography (OCT). It was found that plasma Cer are associated with culprit plaque vulnerability evaluated by OCT, providing evidence supporting proatherogenic roles and potential to act as markers for plaque vulnerability of Cer. Graphical Abstract With increasing plasma ceramide levels, the prevalence of thin-cap fibroatheroma (TCFA) and plaque rupture (PR) is higher, that is, culprit plaques are more vulnerable.

Plasma ceramides are associated with coronary atherosclerotic burden in patients with ST-segment elevation myocardial infarction.

BACKGROUND: Plasma ceramides (Cer), a subset of bioactive lipids, have mechanistic links to atherosclerotic coronary artery disease (CAD) pathogenesis and are related to major adverse cardiovascular events (MACEs). OBJECTIVES: This study aimed to explore the associations between plasma Cer and atherosclerotic burden evaluated by Synergy Between Percutaneous Coronary Intervention with Taxus and Cardiac Surgery (SYNTAX) score. METHODS AND RESULTS: A retrospective series of 248 ST-segment elevation myocardial infarction (STEMI) patients undergoing interventional procedures and plasma ceramides measurement were enrolled. Rapid resolution liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (RRLC-Q-TOF/MS) was used to evaluate plasma Cer concentrations. SYNTAX score was automatically calculated on the SYNTAX website (http://www.syntaxscore.com/). Patients with STEMI had higher plasma MACEs-related ceramide levels than normal donors (p < .001). Pearson correlation analysis showed positive correlations between SYNTAX score and Cer(d18:1/16:0) (r = 0.176, p = .006), Cer(d18:1/18:0) (r = 0.290, p < .001), Cer(d18:1/24:1) (r = 0.209, p = .001) and Cer(d18:1/24:0) (r = 0.134, p = .036). Adjustments for all traditional risks, higher Cer(d18:1/16:0) level (per SD increase, ß (95%CI) =10.681 (1.912-19.923), p = .032), Cer(d18:1/18:0) level (per SD increase, ß (95%CI) =38.830 (15.444-62.126), p = .001), Cer(d18:1/24:1) level (per SD increase, ß (95%CI) =6.122 (1.640-10.605), p = .008) (except for and Cer(d18:1/24:0) level (per SD increase, ß (95%CI) =0.999 (-0.508-2.506), p = .193)) were independently associated with higher levels of SYNTAX score. CONCLUSIONS: Elevated plasma levels of Cer (d18:1/16:0), Cer(d18:1/18:0) and Cer(d18:1/24:1)) are independent predictors for a high atherosclerotic burden in patients with STEMI. Our findings provide evidence supporting proatherogenic roles of Cer.

Relationship between elevated plasma ceramides and plaque rupture in patients with ST-segment elevation myocardial infarction.

BACKGROUND AND AIMS: Ceramides (Cer) are an atherogenic substance. However, the associations between specific plasma Cer levels and culprit plaque morphology in ST-segment elevation myocardial infarction (STEMI) patients are unclear. METHODS: The study consisted of two parallel cohorts. 100 consecutive patients with STEMI were screened as discovery cohort. In the validation cohort, we separately screened 30 normal donors, 30 stable angina pectoris (SAP) and 315 STEMI patients. All STEMI patients underwent emergency percutaneous intervention (PCI) and optical coherence tomography (OCT) examination for culprit plaque. Based on established diagnostic criteria, STEMI patients were classified into plaque rupture (PR) and plaque erosion (PE) group, respectively. Rapid resolution liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (RRLC-Q-TOF/MS) was used to evaluate plasma Cer levels of the screened patients. RESULTS: STEMI patients had higher plasma Cer(d18:1/16:0), Cer(d18:1/18:0), Cer(d18:1/24:1) and Cer(d18:1/24:0) levels than normal donors and SAP patients (p < 0.001, p < 0.001, p < 0.001, p < 0.001, respectively). Plasma Cer levels were significantly higher in STEMI patients with PR than with PE (p < 0.001, p < 0.001, p = 0.008 and p = 0.006, respectively). The frequency of PR increased with increasing tertiles of plasma Cer (adjusted p for trend = 0.004, 0.044, 0.021 and 0.133, respectively). The fully adjusted per SD odds ratios (95% confidence interval) for PR were 9.375 (2.496-35.215) for Cer(d18:1/16:0), 3.586 (1.588-8.089) for Cer(d18:1/18:0), 8.171 (1.764-37.857) for Cer(d18:1/24:1), and 3.831 (1.288-11.289) for Cer(d18:1/24:10). CONCLUSIONS: The study documented novel, positive and independent associations between plasma Cer concentrations and the presence of PR, suggesting plasma Cer may act as potential biomarkers for PR to improve risk stratification.

[Untargeted metabonomics study of ginseng in treatment of spleen-Qi deficiency].

Ginseng has been used to treat Qi-deficiency syndrome up to now, while the therapeutic mechanism is still unclear. In order to explore the mechanism of ginseng in the treatment of Qi-deficiency constitution, the untargeted metabonomics with blood was studied based on rapid resolution high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(RRLC-Q-TOF-MS). In the results, 13 potential biomarkers were found and identified, which mainly involved in the body's antioxidant and immune functions and energy, glycerol, fatty acid, sugar metabolism and bile acid metabolism. The results of blood biochemical analysis also indicated that ginseng could regulate the body's energy metabolism, immune functions and antioxidant capacity in spleen-Qi deficiency constitution. This study revealed the mechanism of ginseng in the treatment of spleen-Qi deficiency using the blood metabonomics, which could provide technological support and scientific basis for further research on ginseng treatment of Qi-deficiency.

Metabolite profiling of Huaiyang Medicago polymorpha with different mowing crops.

The main purpose of our study was to identify and compare secondary metabolites due to different mowing in order to make better use of Huaiyang Medicago polymorpha. The metabolite profiling of Huaiyang Medicago polymorpha with two mowing crops was performed using a rapid resolution liquid chromatography system with quadrupole time-of-flight mass spectrometer (RRLC-QTOFMS) followed by multivariate statistical analyses. Principal Component Analysis (PCA) results showed a clear distinction between two mowing crops. The major metabolites that contributed to mowing discrimination were identified. The results also showed that the content of major active compounds in Medicago polymorpha from the second crop are higher significantly than the first crop. This study suggests that the strategy is a reliable and simple method for the rapid discrimination of subtle variations due to different mowing crops.

Untargeted metabonomics study of ginseng in treatment of spleen-Qi deficiency / 中国中药杂志

Ginseng has been used to treat Qi-deficiency syndrome up to now, while the therapeutic mechanism is still unclear. In order to explore the mechanism of ginseng in the treatment of Qi-deficiency constitution, the untargeted metabonomics with blood was studied based on rapid resolution high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(RRLC-Q-TOF-MS). In the results, 13 potential biomarkers were found and identified, which mainly involved in the body's antioxidant and immune functions and energy, glycerol, fatty acid, sugar metabolism and bile acid metabolism. The results of blood biochemical analysis also indicated that ginseng could regulate the body's energy metabolism, immune functions and antioxidant capacity in spleen-Qi deficiency constitution. This study revealed the mechanism of ginseng in the treatment of spleen-Qi deficiency using the blood metabonomics, which could provide technological support and scientific basis for further research on ginseng treatment of Qi-deficiency.

Simultaneously Quantitative Analysis of Naringin and Its Major Human Gut Microbial Metabolites Naringenin and 3-(4'-Hydroxyphenyl) Propanoic Acid via Stable Isotope Deuterium-Labeling Coupled with RRLC-MS/MS Method.

Widespread in citrus fruits, naringin, a natural 2,3-dihydroflavonoid, is of particular interest to scientists and has a broad range of beneficial bioactivities to health. Orally administered naringin remains in the gut tract for a relatively long time because of its low bioavailability. Under the metabolism mediated by human gut microbiota, naringin could be an active precursor for derived metabolites to play important physiological roles. However, naringin and its metabolites are hard to accurately quantify due to severe endogenic interference. In this study, an analytical rapid resolution liquid chromatography tandem mass spectrometry (RRLC-MS/MS) method coupled with stable isotope deuterium-labeling is developed and validated to simultaneously quantify naringin as well as its major human gut microbial metabolites naringenin and 3-(4'-hydroxyphenyl) propanoic acid. By eliminating the matrix interferences, this strategy not only confirms naringenin and 3-(4'-hydroxyphenyl) propanoic acid as the predominant metabolites which contribute to the pharmacological effects of naringin but also provides a suitable choice for other flavonoid pharmacokinetics study.

Rapid identification and analysis of the active components of traditional Chinese medicine Xiaoxuming decoction for ischemic stroke treatment by integrating UPLC-Q-TOF/MS and RRLC-QTRAP MSn method.

Xiaoxuming decoction (XXMD) is a key Chinese medicine prescription, which has been clinically used for stroke treatment for thousands of years in ancient China. The extracted active fraction of XXMD (AF-XXMD) contains almost pharmacological active components with anti-cerebral ischemic effects. However, the illumination of its complex ingredients remains challenging. In this study, ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC/Q-TOF MS) and rapid resolution liquid chromatography-triple quad linear ion trap mass spectrometry (RRLC-QTRAP MSn) methods were developed for the qualitative and quantitative analysis of AF-XXMD, respectively. Data showed that 48 compounds were identified in AF-XXMD by using UPLC-Q-TOF/MS, including 14 alkaloids, 14 flavonoids, 12 triterpenoids, 3 chromones, 3 monoterpenes, 1 cyanide glycoside, and 1 volatile oil. Among them, 38 components were unambiguously characterized by their reference standards. A total of 15 compounds in AF-XXMD were first reported. Additionally, 33 compounds were quantified by using RRLC-QTRAP MSn in AF-XXMD. This developed RRLC-QTRAP MSn method provides an adequate linearity (r2 > 0.99) and intrabatch and interbatch variations (RSD < 15%), with recovery (60.3%-107.5%) of 33 compounds concerned. The total content of 33 compounds in AF-XXMD reached 31.53%. The high total contents of compounds of Xing Ren, Shao Yao, and Huang Qin in AF-XXMD were 9.52%, 8.85%, and 7.62%, respectively. The data further showed that cyanophoric glycosides, monoterpenes, and flavonoids were the three most abundant components in AF-XXMD. Results provide advantageous information for the comprehensive study of the pharmacokinetic features and pharmacological mechanisms of AF-XXMD.

RRLC-DAD-ESI-MS based and bioactivity guided phytochemical analysis and separation of coumarins from raw extracts of Trigonostemon lutescens.

As extensively active compounds, coumarins are rarely reported on the phytochemistry of the genus Trigonostemon. We herein proposed a fast strategy for analysis and separation of antitumoral active coumarins from the twigs of T. lutescens. Rapid Resolution liquid chromatography coupled with diode array detection and electrospray ionization mass spectrometry (RRLC-DAD-ESI-MS) analysis indicated the existence of coumarins in the twig extracts. Bioactivity guided phytochemical analysis assays revealed that the twig extract contained some active components that significantly inhibited cancer cell viability. Accordingly, a series of coumarins including a new furanocoumarin have been isolated from the twigs of T. lutescens by semi-preparative chromatographic separation. All compounds, especially furan-type coumarins, were reported for the first time from the genus Trigonostemon. The proposed strategy, by combining RRLC-DAD-ESI-MS based and bioactivity guided phytochemical analysis, exemplify a fast method for screening and identifying active components from raw extracts of herbs.

Simultaneous determination of dietary isoprenoids (carotenoids, chlorophylls and tocopherols) in human faeces by Rapid Resolution Liquid Chromatography.

An analytical method was validated for the quantitative determination of isoprenoids compounds in faecal samples, based on liquid-liquid extraction from a small aliquot (0.3-0.5 g of sample) and subsequent analysis by Rapid Resolution Liquid Chromatography (RRLC) on a C30 column. An excellent linear response was observed over the range specified for all dietary isoprenoids, as confirmed by the correlation coefficient, which ranged from 0.9977 to 0.9999. LODs ranged from 0.002 µg to 0.036 µg for lutein and α-tocopherol, respectively. Depending on the compound, LOQs ranged from 0.001 µg (lutein) to 0.120 µg (α-tocopherol). For accuracy testing, spiking of faeces samples with trans-ß-apo-8'-carotenal, α-tocopherol and chlorophyll a were performed (three concentration levels). Excellent recoveries were obtained in all levels (>90%). The intra-day RSD% ranged from 0.86 to 9.78%. The inter-day RSD% was not higher than 10%, except to α-tocopherol (11.34%). In order to assess the applicability of the method faecal samples from a baby fed with different purees formulated from various vegetables were analysed during a six month period. α-carotene, ß-carotene, capsanthin, lycopene, lutein, phytoene, phytofluene, violaxanthin, zeaxanthin and ζ-carotene), and their isomers were identified and quantified using this method. Besides, 2 tocopherols and 9 chlorophylls and derivatives were identified and quantified in the faecal samples analysed. This method is suitable to determine dietary isoprenoids from complex matrices such as human faeces within 28 min.

Pharmacokinetics and distribution of schisandrol A and its major metabolites in rats.

1. Schizandrol A is an active component in schisandra, also the representative component for the identification of schisandra. 2. A rapid resolution liquid chromatography coupled with quadruple-time-of-flight mass spectrometry (RRLC-QTOF/MS) was developed to investigate the pharmacokinetics of schizandrol A after its intragastric administration (50 mg/kg) in rats. 3. Schizandrol A was rapidly absorbed (T max = 2.07 h), with a longer duration (t 1/2 = 9.48 h) and larger apparent volume of distribution (Vz/F = 111.81 l/kg) in rats. Schizandrol A can be detected in main organs and the order of its distribution was in the liver > kidney > heart > spleen > brain, particularly higher in the liver. 4. Five schizandrol A metabolites were identified, including 2-demethyl-8(R)-hydroxyl-schizandrin, 3-demethyl-8(R)-hydroxyl-schizandrin, hydroxyl-schizandrin, demethoxy-schizandrin, 2, 3-demethyl-8(R)-hydroxyl-schizandrin, indicating that the hydroxylation and demethylation may be the major metabolic way of schizandrol A. 5. This study defined the pharmacokinetic characteristics of schizandrol A in vivo, and the RRLC-QTOF/MS is more sensitive and less limited by conditions, and needs less samples, which may be a useful resource for the further research and development of schisandrol A.

[Rapid analysis on chemical constituents in Qige Keli by RRLC-Q-TOF-MS/MS].

The qualitative analysis method of RRLC-Q-TOF-MS/MS was established for determine the chemical constituents in Qige Keli. Kramosil C18 column (4.6 mm×150 mm, 3.5 µm) was used at the temperature of 30 °C. The mobile phase was 0.2% formic acid and acetonitrile by gradient elution, with a flow rate at 1.0 mL·min⁻¹, and the injection volume was 10 µL. The high-resolution quadrupole time-flight mass spectrometry was used as detector with electrospray ion source in both positive and negative models. On the basis of medicinal materials, reference materials, literature reports, and mass spectrometry data, the chemical composition in the Qige Keli was identified. A total of 44 compounds were identified, including 3 flavonoids, 21 flavonoid glycosides, 8 organic acids, 6 lactones, and 3 saponins. The results laid the foundation for the quality control of Qige Keli and the further research on pharmacodynamic materials.

[Metabolic and excretory process of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium in rats' urine and feces analyzed by RRLC-QqQ-MSn].

This present study was to investigate the metabolism and excretion of characteristic polyphenols such as flavonoids and coumarins in urine and feces of rats after intragastric administration of ethanol extracts of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium. The urine and feces of rats were collected after intragastric administration of 70% ethanol extracts of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium. Rapid resolution liquid chromatography coupled with quadrupole tandem mass spectrometry (RRLC-QqQ-MSn) was applied to compare the contents of polyphenols in ethanol extract, urine and feces. By comparing with reference substance, 30 polyphenols were identified from the ethanol extracts of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium, including flavone glycosides, flavones, flavonone glycosides, flavonones, flavonol glycosides, polymethoxyflavones, coumarins, and limonoids and so on. The detection of various types of compounds showed differences in contents between the intestinal metabolism and excretion in the feces after systemic circulatory metabolism and renal excretion. The results showed that the polymethoxyflavones and flavonones were primarily excreted through urine, and the flavonone glycosides and limonoids were primarily excreted through feces. However, coumarins were hardly detected in feces and urine, indicating that coumarins may be metabolized in the body.

[Determination of flavonoids constituents in epimedium total flavone capsule and research on its anti-osteoporosis activity].

To develop a rapid resolution liquid chromatography (RRLC) method for the simultaneous determination of epimedoside A, epimedin A1, epimedin A, epimedin B, epimedin C, icariin, baohuosideⅡ, icarisideⅠ, sagittatoside B, 2"-O-rhamnosyl icarisideⅡ, and baohuosideⅠin epimedium total flavone capsule. At the same time, the effects of the above 11 compounds on osteogenic differentiation of MC3T3-E1 cells were investigated by detecting the content of alkaline phosphatase (AKP). The results showed that baohuoside Ⅱ had the highest activities, and both the activities of baohuoside Ⅱ and icariside Ⅰ were stronger than those of icariin.In this study, the content determination method of flavonoid glycosides was established, and the anti-osteoporosis effect of monomers was compared, providing technical support for the study of the pharmacodynamic and mechanism of Epimedium total flavone capsule.

Microdialysis combined with RRLC-MS/MS for the pharmacokinetics of two major alkaloids of Bi qi capsule and the potential roles of P-gp and BCRP on their penetration.

Bi qi capsule (BQC) is a traditional Chinese medicine prescription that is clinically used for the treatment of rheumatoid arthritis. Strychnine and brucine, as two typical kinds of alkaloids, are the primary active and neurotoxic constituents of BQC. In this study, a sensitive and reliable rapid resolution liquid chromatography-tandem mass spectrometry (RRLC-MS/MS) quantitative method was used to determine the concentrations of brucine and strychnine in rat brain and blood dialysates. The blood-brain barrier (BBB) penetration of free brucine and strychnine and their pharmacokinetic characteristics were investigated by the validated RRLC-MS/MS method coupled with in vivo microdialysis for the first time. The dialysate brain-blood AUC ratios of brucine were 0.098, 0.44 and 0.40 respectively at 0.4, 0.8 and 1.6 g kg-1 doses of BQC, and the dialysate brain-blood AUC ratios of strychnine were 0.20, 1.25 and 2.06 respectively at 0.4, 0.8 and 1.6 g kg-1 doses of BQC. The high brain-blood AUC ratios of brucine and strychnine were observed in medium and high dose groups of BQC. In addition, the effects of P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) on brucine and strychnine across BBB were also studied using the above method as well as molecular docking. The results prompted that brucine was the substrate of P-gp, and strychnine might be the inhibitor of P-gp. Brucine and strychnine showed high brain penetration, so it is very important to well control the clinic dosage of BQC and manufactory quality for avoiding the side effects and obtaining good therapeutic efficacy. Our study could be further used in investigating BBB penetration for other drugs caused neurotoxicity.

Metabolomics study of the therapeutic mechanism of Schisandra chinensis lignans on aging rats induced by d-galactose.

OBJECTIVE: The aim of this study was to evaluate the antiaging effect of Schisandra chinensis lignans (SCL) by analyzing the characteristics in the serum of d-galactose (d-gal)-induced rats. METHODS: Forty male Wistar rats were randomly divided into control group, d-gal model group, low-dose SCL group (50 mg/kg/d), medium-dose SCL group (100 mg/kg/d), and high-dose SCL group (200 mg/kg/d). A serum metabolomics analysis method based on rapid resolution liquid chromatography coupled with quadruple-time-of-flight mass spectrometry was carried out to study the characteristics of d-gal-induced aging rats and evaluate the antiaging effects of SCL, and multivariate statistical analysis was performed for pattern recognition and characteristic metabolites identification. The relative levels of p19, p53, and p21 genes in the brain tissue were measured by quantitative real-time polymerase chain reaction for investigating the underlying mechanism. RESULTS: Metabolomics analysis showed that 15 biomarkers were identified and 13 of them recovered to the normal levels after the administration of SCL. Based on the pathway analysis, the antiaging mechanisms of SCL might be involved in the following metabolic pathways: energy, amino acid, lipid, and phospholipid metabolism. Furthermore, SCL significantly inhibited the mRNA expression level of p19, p53, and p21 in the brain of aging rats induced by d-gal. CONCLUSION: These results suggest that SCL can delay rat aging induced by d-gal through multiple pathways.

Anti-ageing effect of red ginseng revealed by urinary metabonomics using RRLC-Q-TOF-MS.

INTRODUCTION: Red ginseng (RG) is one of the main processed products of the roots and rhizomes of Panax ginseng C.A. Meyer and is used for anti-ageing. But how metabonomic influences of RG on the progress of ageing are less researched. OBJECTIVE: A metabonomic method was developed to study the characters of the ageing process and the effects of total ginsenosides of red ginseng (TGRG) on the progress of ageing. METHODS: Urine samples from four different ages (4, 12, 18 and 24 months old) of rats and interference after TGRG were analysed by rapid resolution liquid chromatography coupled with quadruple-time-of-flight mass spectrometry (RRLC-Q-TOF-MS) and multivariate statistical analysis were performed for the pattern recognition and characteristic metabolites identification. RESULTS: Fourteen potential biomarkers were found and identified by MS/MS analysis by referring to authentic chemicals. The analysis of metabolic pathways indicated that the reduced energy and lipid metabolism, the decline of kidney function and amino acids metabolism disorders were the main features of ageing. After TGRG administration, lipid and amino acids metabolism of 18 and 24 month-old rats were adjusted to restore a younger level, and nine related biomarkers in the ageing process reset to a younger level were recognised. CONCLUSION: These changes showed that TGRG may produce an anti-ageing effect by intervening in the lipid metabolism and correcting the amino acid metabolism disorders in ageing rats.

Plasma metabolomics reveal the correlation of metabolic pathways and Prakritis of humans.

BACKGROUND: Ayurveda, an ancient Indian medicinal system, has categorized human body constitutions in three broad constitutional types (prakritis) i.e. Vata, Pitta and Kapha. OBJECTIVES: Analysis of plasma metabolites and related pathways to classify Prakriti specific dominant marker metabolites and metabolic pathways. MATERIALS AND METHODS: 38 healthy male individuals were assessed for dominant Prakritis and their fasting blood samples were collected. The processed plasma samples were subjected to rapid resolution liquid chromatography-electrospray ionization-quadrupole time of flight mass spectrometry (RRLC-ESI-QTOFMS). Mass profiles were aligned and subjected to multivariate analysis. RESULTS: Partial least square discriminant analysis (PLS-DA) model showed 97.87% recognition capability. List of PLS-DA metabolites was subjected to permutative Benjamini-Hochberg false discovery rate (FDR) correction and final list of 76 metabolites with p < 0.05 and fold-change > 2.0 was identified. Pathway analysis using metascape and JEPETTO plugins in Cytoscape revealed that steroidal hormone biosynthesis, amino acid, and arachidonic acid metabolism are major pathways varying with different constitution. Biological Go processes analysis showed that aromatic amino acids, sphingolipids, and pyrimidine nucleotides metabolic processes were dominant in kapha type of body constitution. Fat soluble vitamins, cellular amino acid, and androgen biosynthesis process along with branched chain amino acid and glycerolipid catabolic processes were dominant in pitta type individuals. Vata Prakriti was found to have dominant catecholamine, arachidonic acid and hydrogen peroxide metabolomics processes. CONCLUSION: The neurotransmission and oxidative stress in vata, BCAA catabolic, androgen, xenobiotics metabolic processes in pitta, and aromatic amino acids, sphingolipid, and pyrimidine metabolic process in kapha Prakriti were the dominant marker pathways.

Toxicokinetics of strychnine and brucine after the oral administration of Biqi capsule to rats by RRLC-MS/MS.

Biqi capsule is a well-known traditional Chinese medicine formula that has been widely applied for the clinical treatment of such diseases as rheumatoid arthritis, scapulohumeral periarthritis and cervical spondylopathy. However, there is concern regarding the toxicity of Biqi capsule owing to its active ingredients, strychnine and brucine. To investigate the toxicokinetics of strychnine and brucine after oral administration of Biqi capsule to rats, a sensitive and simple rapid-resolution liquid chromatography/tandem mass spectrometry method was developed to determine the levels of strychnine and brucine in rat plasma. Chromatographic separation was performed on a Capcell Pak C18 MG II (3.0 µm, 2.0 × 35 mm) column by gradient elution with acetonitrile and 0.2% formic acid as the mobile phase. The method was validated over the range of 0.25-250 ng/mL for strychnine and 0.025-25 ng/mL for brucine. The intra- and inter-day accuracies of strychnine and brucine in rat plasma were 100.3-106.6 and 90.75-106.1% respectively, and the precisions were within 14.2%. The established method was successfully applied to the toxicokinetic study of strychnine and brucine after single and multiple oral administration of Biqi capsule to male and female rats at 0.4, 0.8 and 1.6 g/kg doses. The results showed different toxicokinetic characteristics in the different groups.