Rapid Resolution Liquid Chromatography Research Articles

Dissipation behavior and dietary risk assessment of lambda-cyhalothrin, thiamethoxam and its metabolite clothianidin in apple after open field application

The dissipation dynamics and residue amounts of lambda-cyhalothrin, thiamethoxam and clothianidin in apple were investigated by using rapid resolution liquid chromatography triple quadrupole mass spectrometer (RRLC-MS/MS) and gas chromatography mass spectrometry (GC-MS). The developed method performed satisfactory recoveries of 88%–105% and the limit of quantitation (LOQ) was 0.01 mg kg−1. The suspension concentrate (SC) formulation of lambda-cyhalothrin and thiamethoxam was applied on apple field in accordance with good agricultural practice (GAP). The half-lives of two pesticides ranged from 7.01 d to 17.3 d and the terminal residues were <0.01–0.21 mg kg−1. Based on the Chinese dietary pattern, the dietary risk of lambda-cyhalothrin and total thiamethoxam were predicted by comparing intake amounts with the toxicological data, namely acceptable daily intake (ADI) and acute reference dose (ARfD). The chronic and acute risk quotients were 0.1080–0.4463 and 0.0008–0.2005, respectively, which showed negligible risk for general consumers. The pre-harvest interval (PHI) of 21 d was suggested for the formulation in compliance with maximum residue limit (MRL) and dietary risk assessment, meanwhile, the MRL of 0.1 mg kg−1 was recommended for thiamethoxam in apple. These results were vital for guiding reasonable usage of two insecticides and for approval of formulation use.

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-β-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, β-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.

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.

Dissipation behavior, residue distribution and dietary risk assessment of field-incurred boscalid and pyraclostrobin in grape and grape field soil via MWCNTs-based QuEChERS using an RRLC-QqQ-MS/MS technique

A QuEChERS technique employing multi-walled carbon nanotubes was developed and validated for the simultaneous quantitation of boscalid and pyraclostrobin in grape field samples using rapid resolution liquid chromatography triple- quadrupole tandem mass spectrometry. The validated method was successfully applied to determine the levels of residual boscalid and pyraclostrobin in soil and grapes. The dissipation behavior of each fungicide followed first-order kinetics, with a linear correlation coefficient of 0.995–0.998 at 95% confidence. The highest levels of boscalid and pyraclostrobin in grape, with a pre-harvest interval of 14 days and doses of 380–570 mg a.i. kg−1 in three or four applications, were 3.99 mg kg−1 and 0.792 mg kg−1, respectively. These are below the maximum residue limits for boscalid and pyraclostrobin in grape, as recommended by the Chinese Ministry of Agriculture. Risk quotients (RQs) of these pesticides were evaluated by comparing national estimated daily intake with acceptable daily intake. The results yielded RQs for boscalid and pyraclostrobin in grape of 54% and 31.7%, respectively, which suggest a low health risk to consumers.

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.

Identification and Evaluation of a Panel of Ginsenosides from Different Red Ginseng Extracts with Nootropic Effect

Red ginseng has been gradually discovered to have pharmacological and physiological effects. It is well known that the most important bioactive components of ginseng are ginsenosides. The nootropic effect of ginsenosides from nine different red ginseng extracts was evaluated here. Nine groups of mice were perfused with different concentrations of nine red ginseng extracts, respectively, and two groups of mice with distilled water. The nootropic effect of ginsenosides on mice was evaluated with behavior tests and a biochemical indicator study. The extracts were identified by rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrome-try(RRLC-Q-TOF-MS). Furthermore, principal component analysis(PCA) was used to analyze the contribution of chemical components from different ginseng groups. The extracts with the most and the weakest effective nootropic were found. It is notable that extract processing is a very important factor to decide pharmacological functions of ginseng extracts. As a conclusion, the most effective extract method for ginsenosides has been found. A panel of 13 ginsenosides has been screened out as chemical markers with nootropic effect, which include high level ginsenosides Ra0, Rb1, Rc, Rb2, Rb3, Re, Rd, and Rg1 and low level ginsenosides mRb1, mRc, mRb2, mRd, and F2. Low level ginsenosides were first time to be discovered as possible nootropic compounds. This method may shed light on fast discovery of bioactive compounds of medicinal plants with low level compounds.

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

ObjectiveThe 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.MethodsForty 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.ResultsMetabolomics 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.ConclusionThese results suggest that SCL can delay rat aging induced by d-gal through multiple pathways.

Plasma trimethylamine N-oxide is associated with vulnerable plaque characteristics in CAD patients as assessed by optical coherence tomography

BackgroundPlaque vulnerability indicates the risk of a cardiovascular event. In the present study, we sought to analyze the relationship between trimethylamine N-oxide (TMAO), a gut microbiota metabolite from dietary phosphatidylcholine, and vulnerable plaque characteristics in patients with coronary artery disease (CAD). MethodsOne hundred eighty non-culprit plaques from 90 patients with ACS or with stable angina were assessed by optical coherence tomography (OCT). The plasma TMAO levels were measured using rapid resolution liquid chromatography quadrupole time-of-flight mass spectrometry (RRLC-QTOF/MS). ResultsPatients were divided into two groups (high TMAO group and low TMAO group) according to the median plasma TMAO level (114.73 μg/L). The non-culprit plaques in the high TMAO group exhibited a thinner fibrous cap thickness (FCT) (65.97 ± 25.89 vs. 93.0 ± 28.28 μm, P < 0.001), higher frequency of microvessels (75.6% vs. 31.1%, P < 0.001, per-patient) and increased incidence of thin-cap fibroatheroma (TCFA) (69.2% vs. 18.4%, P < 0.001, per-patient) compared with the low TMAO group. Moreover, the level of TMAO was negatively associated with FCT (r = −0.418, P < 0.001). Furthermore, multivariate regression analysis results showed that TMAO (OR: 7.455, 95% CI: 2.753–20.189, P < 0.001) had a significant association with TCFA, with a cut-off point of 118.34 μg/L, specificity of 72.6% and sensitivity of 79.5% in predicting the prevalence of TCFA. ConclusionsIn conclusion, these findings suggest that the level of TMAO is significantly correlated with the incidence of TCFA. New biomarkers acquired through non-invasive means, such as TMAO, offer the potential to improve risk stratification and clinical management in patients with CAD.

Simultaneous determination of rosuvastatin, naringin and naringenin in rat plasma by RRLC-MS/MS and its application to a pharmacokinetic drug interaction study.

A rapid resolution liquid chromatography tandem mass spectrometry method was developed and validated for simultaneous determination of rosuvastatin, naringin and naringenin in rat plasma. Chromatographic separation of analytes and internal standard (fluvastatin for rosuvastatin, while isoquercitrin for naringin and naringenin) was performed on Agilent Poroshell 120 EC-C18 column (3.0 × 50 mm, 2.7 μm) using gradient elution with a mobile phase of methanol and water, both with 0.1% formic acid (v/v). The detection was operated in multiple reaction monitoring mode to monitor the precursor-to-product ion transitions of m/z 579.1→270.8 for naringin, m/z 270.9→150.7 for naringenin, m/z 463.1→299.8 for isoquercitrin in negative ionization mode, and m/z 482.2→258.1 for rosuvastatin, m/z 412.1→224.1 for fluvastatin in positive ionization mode. Polarity switch (negative-positive-negative ionization mode) was performed in a total runtime of 5.0 min. The method was validated over a concentration range of 10-2,000 ng/mL for the above three analytes. The intra-day and inter-day precisions and accuracies of the quality control samples at low, medium and high concentration levels exhibited relative standard deviations <10% and the accuracy values ranged from -7.2% to 8.4%. The proposed method was successfully applied to the pharmacokinetic drug interaction study of rosuvastatin combined with naringin in rats.

Pharmacokinetic and metabolic studies of ginsenoside Rb3 in rats using RRLC-Q-TOF-MS.

Ginsenoside Rb3 is one of major ginsenosides in Panax ginseng with effect on cardio-vascular and central nervous system. The aim of this study is to develop a rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (RRLC-Q-TOF-MS) method for pharmacokinetic study of ginsenoside Rb3 and simultaneous determination of metabolites in rats. The results showed that the concentration-time profile of ginsenoside Rb3 conformed to a two-compartment pharmacokinetic model after intravenous administration at the dosage of 2.0 mg/kg for rats. The mean plasma elimination half-lives were 13.77 ± 1.23 min and 2045.70 ± 156.20 min for the distribution and exterminate phases t1/2α and t1/2β. In the metabolic study, prototype ginsenoside Rb3 and deglycosylation metabolites were characterized by comparison with the retention time of the standard compounds, accurate mass measurement and the characteristic fragment ions obtained from MS/MS. Two major metabolites Mb1 and M2' were tentatively identified in rat urine samples after intravenous administration, and four possible metabolites Mb1, F2, M2' and CK were detected in rat feces samples after oral administration. The deglycosylation was found to be the major metabolic pathways of ginsenoside Rb3 in rat. The in vivo metabolic pathway of ginsenoside Rb3 was summarized.

Sensitive Rapid Fluorescence Polarization Immunoassay for Free Mycophenolic Acid Determination in Human Serum and Plasma.

In this Article, we describe a fluorescence polarization immunoassay (FPIA) using a new label-near-infrared fluorescent dye. The developed FPIA method was optimized for the rapid analysis of free mycophenolic acid (MPA) in plasma of transplanted patients. The approach is based on the fluorescence competitive assay between the target immunosuppressant and a novel emissive near-infrared fluorescent dye-tagged MPA and MPA-AO for the binding sites of the anti-MPA antibody. The fluorescent analogue of MPA exhibits emission at 654 nm upon excitation at 629 nm (λexcmax) and shows a good photochemical stability and a significant emission quantum yield (0.16) in phosphate buffer media. Free mycophenolic acid was isolated from blood or plasma samples using ultrafiltration prior to analysis. The sample was incubated for 20 min with 5 μg/mL of anti-MPA antibody and 1 nM of MPA-AO before the measurements. The developed FPIA displays a limit of detection of 0.8 ng/mL (10% binding inhibition) and a dynamic range of 1.7-39 ng/mL (20%-80% binding inhibition) in a PBST buffer, fitting the therapeutic requirements. The immunoassay selectivity was evaluated by measuring the cross-reactivity to other immunosuppressive drugs administered in combination with MPA (cyclosporin A and tacrolimus), as well as for the metabolite MPA glucuronide. The assay has been successfully applied to the analysis of free MPA in the blood of a heart-transplanted patient after oral administration of both mycophenolate mofetil (MMF) and tacrolimus, and the results have been compared with those obtained by rapid-resolution liquid chromatography with diode array detection (RRLC-DAD).

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

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. 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. Urine samples from four different ages (4, 12, 18 and 24months 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. 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 24month-old rats were adjusted to restore a younger level, and nine related biomarkers in the ageing process reset to a younger level were recognised. 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.

Dissipation behavior, residue distribution and dietary risk assessment of chlorfenapyr and clothianidin in leek using RRLC-QqQ-MS/MS technique

Commercial suspension emulsion (SE) of (8% chlorfenapyr+20% clothianidin), as a pre-registered product in China, was firstly investigated under Chinese open-field conditions. A MWCNTs-based QuEChERS method for simultaneous determination of chlorfenapyr and clothianidin in leek was established and validated through rapid resolution liquid chromatography triples quadrupole tandem mass spectrometry (RRLC-QqQ-MS/MS). Based on this method, the dissipation behaviors, residue distributions and dietary risk probability of these fungicides in leek from two representative locations in China in 2016, which were previously treated with these insecticides at the doses of 420–630g a.i./ha twice applications, were further investigated for food safety. Dissipation behavior of chlorfenapyr and clothianidin in leek followed first-order kinetics with the half-lives of 2.9–7.2days, and the highest residues (HR) of these insecticides in leek were below 0.3811mg/kg and 0.2989mg/kg, respectively, at the pre-harvest interval (PHI, 7days). For dietary risk assessments, the risk quotients (RQs) of chlorfenapyr and clothianidin in leek were evaluated by comparing the value of national estimated daily intake (NEDI) with acceptable daily intake (ADI), based on the trials data under good agricultural practices (GAP) conditions. The results indicated that RQs of chlorfenapyr and clothianidin in leek were 21.5% and 0.29%, respectively, which exhibited an acceptably low health risk to leek consumption. The current study could not only guide reasonable usage of the formulation, but also facilitate the setting of maximum residue limits (MRLs) of chlorfenapyr and clothianidin in leek for Chinese authorities

RAPID RESOLUTION LIQUID CHROMATOGRAPHY METHOD FOR DETERMINATION OF CHLOROGENIC ACID IN ECHINACEA EXTRACTS

This study presents a development and validation of a new, fast, efficient and cost effective reversed-phase rapid resolution liquid chromatography (RP-RRLC) method for determination of chlorogenic acid in echinacea extracts. The optimum separation with symmetrical peak shape and good index purity of the analyte was achieved on a Poroshell 120 EC-18 (50 mm x 3 mm; 2.7 µm) analytical column, mobile phase consisted of acetonitrile/(water with 1 % phosphoric acid), (10/90, V/V)in isocratic elution with flow rate of 1 mL/min and UV diode-array detection (UV-DAD) at 325 nm. The developed method was validated by testing specificity, selectivity, linearity, precision, accuracy, limit of detection (LOD) and limit of quantification (LOQ). The calibration curve of chlorogenic acid followed Beer’s law within the range 28.97 ng - 362.19 ng (R2 = 0.9994). The LOD was 0.29 pg, while LOQ was 0.96pg. The intra-day precisions was evaluated for the retention time, peak area and peak height and the calculated values for relative standard deviations (RSD) were 0.21 %, 0.11 % and 0.22 %, respectively. The mean recoveries ranged from 98.75 to 104.63 % and RSD was less than 0.23 %. The developed method was successfully applied for identification and quantification of chlorogenic acid in three samples of echinacea extracts, taken from local pharmacies.

Simultaneous determination of pyridaben, dinotefuran, DN and UF in eggplant ecosystem under open-field conditions: Dissipation behaviour and residue distribution

A sensitive method for simultaneous determination of pyridaben, dinotefuran, DN and UF in eggplant ecosystem was established and validated through rapid resolution liquid chromatography triples quadrupole tandem mass spectrometry (RRLC-QqQ-MS/MS). Matrix-matched external calibrations were introduced to check matrix effects. Limits of quantification (LOQs) of pyridaben, dinotefuran, DN and UF in eggplant were 0.2, 0.2, 1.0 and 1.0 μg kg−1, and 0.2, 0.2, 5.0 and 1.0 μg kg−1 in soil, respectively. Limits of detection (LODs) of four pesticides were below 0.41 μg L−1. The mean recoveries (n = 5) of these insecticides varied from 79.4% to 103%, and the relative standard deviations (RSDs) ranged from 2.1% to 15.3% at three levels. This method was applied to Chinese open-field samples from two representative locations, which were previously treated with these insecticides at the doses of 210–315 g a.i. ha−1 twice or three times. The dissipations of pyridaben and dinotefuran in eggplant and soil followed first-order kinetics with the half-lives of 3.65–11.4 d. The residues of pyridaben and total dinotefuran (calculated as sum of dinotefuran parent, DN and UF) in eggplant were below 0.0311 mg kg−1 at the pre-harvest interval (PHI, 7 d). Presently, no maximum residue limit (MRL) of pyridaben and dinotefuran in eggplant was recommended by China, Codex Alimentarius Commission (CAC) or European Union (EU). This study was important for evaluation of environmental fate and food safety of pyridaben and dinotefuran in eggplant ecosystems, and facilitated China to establish maximum residue limits (MRLs) of pyridaben and dinotefuran in eggplant.

Simultaneous Determination of Pyraclostrobin, Prochloraz, and its Metabolite in Apple and Soil Via RRLC-MS/MS

A sensitive and high-efficiency method for simultaneous detection of pyraclostrobin, prochloraz, and its metabolite in apple and soil based on QuEChERS pretreatment combined with rapid resolution liquid chromatography tandem mass spectrometry was established and validated. The limits of quantification of three compounds in two matrixes were 0.005 mg kg−1. The average recoveries of pyraclostrobin, prochloraz, and 2,4,6-trichlorophenol in soil matrix were in the ranges of 87–105%, 86–95%, and 90–96%, respectively, with all the relative standard deviations (RSDs) of < 9.6%, while recoveries were 89–93%, 83–97%, and 89–101% in apple with the RSDs of < 6.5% at three spiking levels. For verify the applicability of this method, the real samples from three representative locations were detected. The degradation behaviors and residue distributions of pyraclostrobin and prochloraz and its metabolite in apple ecosystem were investigated. The field experiment data showed that the dissipation of pyraclostrobin and prochloraz in apple and soil followed pseudo-first-order kinetic models. The half-lives of pyraclostrobin in soil and apple were 8.6–19.8 and 7.9–15.1 days, while prochloraz were 8.9–21 and 5.8–12.4 days. The highest terminal residue of total prochloraz and pyraclostrobin in apple, after spraying three to four times with the interval of 28 days, was far below the maximum residue limits recommended by China. This research could provide guidance on a reasonable usage of prochloraz and pyraclostrobin in apple orchard.

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

BackgroundAyurveda, an ancient Indian medicinal system, has categorized human body constitutions in three broad constitutional types (prakritis) i.e. Vata, Pitta and Kapha. ObjectivesAnalysis of plasma metabolites and related pathways to classify Prakriti specific dominant marker metabolites and metabolic pathways. Materials and methods38 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. ResultsPartial 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. ConclusionThe 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.

Comparing eight types of ginsenosides in ginseng of different plant ages and regions using RRLC-Q-TOF MS/MS

BackgroundThis article aims to compare and analyze the contents of ginsenosides in ginseng of different plant ages from different localities in China. MethodsIn this study, 77 fresh ginseng samples aged 2–4 years were collected from 13 different cultivation regions in China. The content of eight ginsenosides (Rg3, Rc, Rg1, Rf, Rb2, Rb1, Re, and Rd) was determined using rapid resolution liquid chromatography coupled with quadrupole–time-of-flight tandem mass spectrometry (RRLC-Q-TOF MS/MS) to comparatively evaluate the influences of cultivation region and age. ResultsGinsenoside contents differed significantly depending on age and cultivation region. The contents of ginsenosides Re, Rc, Rg1, Rg3, and Rf increased with cultivation age, whereas that of ginsenoside Rb1 peaked in the third year of cultivation. Moreover, the highest ginsenoside content was obtained from Changbai (19.36 mg/g) whereas the lowest content was obtained from Jidong (12.05 mg/g). Ginseng from Jilin Province contained greater total ginsenosides and was richer in ginsenoside Re than ginseng of the same age group in Heilongjiang and Liaoning provinces, where Rb1 and Rg1 contents were relatively high. ConclusionIn this study, RRLC-Q-TOF MS/MS was used to analyze ginsenoside contents in 77 ginseng samples aged 2–4 years from different cultivation regions. These patterns of variation in ginsenoside content, which depend on harvesting location and age, could be useful for interested parties to choose ginseng products according to their needs.

Simultaneous Quantification of a Herbal Combination of Pueraria lobata, Salvia miltiorrhiza and Panax notoginseng by Rapid Resolution Liquid Chromatography

Pueraria lobata, Salvia miltiorrhiza and Panax notoginseng are traditional Chinese medicines which have been used for thousands of years either singly or in combination. However, the quality of formulas containing either all or part of the three herbs is, at present, controlled by only one or two easily detectable compounds.. A rapid resolution liquid chromatography (RRLC) method has been developed for the quality control of the three herbs in combination through ten active ingredients originating from the raw herbs. The analysis was achieved in a run of less than 25 minutes. Using the same method, the finger-prints of the three herbs are detectable in the chromatogram, and so the method could be applied for identification of different species in the same genus. Four formula preparations {xinkeshu capsule (XKSC), xinkeshupian (XKSP), fufang-danshenpian (FFDSP), and naodeshengpian (NDSP)} were analyzed by this new RRLC method. Controlling only one or two easily detectable ingredients is far from enough to assure the quality of the formula preparations. However, controlling multiple ingredients originating from the raw herbs and their ratios is an effective and feasible approach to solve this problem.

Metabolomics study of the therapeutic mechanism of Schisandra Chinensis lignans in diet-induced hyperlipidemia mice

BackgroundSchisandra, a globally distributed plant, has been widely applied for the treatment of diseases such as hyperlipidemia, fatty liver and obesity in China. In the present work, a rapid resolution liquid chromatography coupled with quadruple-time-of-flight mass spectrometry (RRLC-Q-TOF-MS)-based metabolomics was conducted to investigate the intervention effect of Schisandra chinensis lignans (SCL) on hyperlipidemia mice induced by high-fat diet (HFD).MethodsHyperlipidemia mice were orally administered with SCL (100 mg/kg) once a day for 4 weeks. Serum biochemistry assay of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-c) and high-density lipoprotein cholesterol (HDL-c) was conducted to confirm the treatment of SCL on lipid regulation. Metabolomics analysis on serum samples was carried out, and principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) were carried out for the pattern recognition and characteristic metabolites identification. The relative levels of critical regulatory factors of liver lipid metabolism, sterol regulatory element-binding proteins (SREBPs) and its related gene expressions were measured by quantitative real-time polymerase chain reaction (RT-PCR) for investigating the underlying mechanism.ResultsOral administration of SCL significantly decreased the serum levels of TC, TG and LDL-c and increased the serum level of HDL-c in the hyperlipidemia mice, and no effect of SCL on blood lipid levels was observed in control mice. Serum samples were scattered in the PCA scores plots in response to the control, HFD and SCL group. Totally, thirteen biomarkers were identified and nine of them were recovered to the normal levels after SCL treatment. Based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis, the anti-hyperlipidemia mechanisms of SCL may be involved in the following metabolic pathways: tricarboxylic acid (TCA) cycle, synthesis of ketone body and cholesterol, choline metabolism and fatty acid metabolism. Meanwhile, SCL significantly inhibited the mRNA expression level of hepatic lipogenesis genes such as SREBP-1c, fatty acid synthase (FAS) and acetyl-CoA carboxylase (ACC), and decreased the mRNA expression of liver X receptor α (LXRα). Moreover, SCL also significantly decreased the expression level of SREBP-2 and 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR) in the liver of hyperlipidemia mice.ConclusionAnti-hyperlipidemia effect of SCL was confirmed by both serum biochemistry and metabolomics analysis. The mechanism may be related to the down-regulation of LXRα/SREBP-1c/FAS/ACC and SREBP2/HMGCR signaling pathways.