Rapid Resolution Liquid Research Articles

Quality Control of the Fuzi Lizhong Pill Through Simultaneous Determination of 16 Major Bioactive Constituents by RRLC-MS-MS.

Fuzi Lizhong pill (FLP) is used to treat gastritis, and the monarch drug of it is Aconiti Lateralis Radix Praeparata (Fuzi, aconite roots) which is a toxic herbal medicine. To better control the safety and quality of FLP, an effective method to analyze the contents of 16 toxic and bioactive components using rapid resolution liquid chromatography-tandem triple-quadrupole mass spectrometer was established. The 16 constituents included aconine, mesaconine, hypaconitine, benzoylaconine, benzoylmesaconine, benzoylhypaconine, adenosine, liquiritin, liquiritigenin, glycyrrhizic acid, isoliquiritigenin, 6-gingerol, atractylenolide III, atractylenolide I, atractylenolide II and glycyrrhetic acid. Ideal separation was performed using gradient elution in 13 min by optimized conditions. All the isomerides were isolated to baseline. The improved method with a polarity switch in contiguous time segments could analyze the five types of components, including polar and nonpolar compounds, without decreasing sensitivity. The proposed method was fully validated. The results revealed that contents of six alkaloids from Fuzi were significantly different among the samples. Using the established method and multivariate statistical method, the quality consistency of two dosage forms of FLP from different companies were analyzed. The optimized method could be used for the quality control of FLP and investigate index compound variation between two dosage forms.

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.

Quality control of Guci tablets using UPLC-ELSD fingerprint analysis coupled with chemometrics

Ultra-performance liquid chromatography-evaporative light scattering detection (UPLC-ELSD) fingerprint analysis method was established for quality control of Guci tablets. Chromatographic separation was performed on Waters Acquity UPLC BEH C₁₈ column (2.1 mm×100 mm, 1.7 μm) at 30 °C of column temperature. Acetonitrile-0.1% formic acid solution was adopted as mobile phase for gradient elution. The flow rate was set at 0.3 mL·min⁻¹, and the injection volume was 3 μL. Detection was carried out on an ELSD with a nitrogen pressure of 0.28 MPa, drift tube temperature of 60 °C, and gain of 400. A total of 39 batches of samples produced by six manufacturers were measured by using the above method and the data were analyzed by ChemPattern software. The peak present in more than 75% of the samples was defined as a common peak, and 30 common peaks were determined. Among them, 19 peaks were identified by rapid resolution liquid chromatography/tandem mass spectrometry (RRLC-MS/MS) method, 16 of which were confirmed by reference substances. The similarity of the tested samples was 0.47-0.98, suggesting that the quality of the samples from different manufacturers varied greatly. Furthermore, principal component analysis (PCA) and hierarchical analysis (HCA) were performed to clarify the main different components in samples. The results indicated that there might be some feeding problems about Paeoniae Radix Alba, Notoginseng Radix et Rhizoma, and Clematidis Radix et Rhizoma in a few manufacturers. This study provided some evidences for the overall quality control of Guci tablets, as well as its quality standard improvements.

Antibiotics in Crab Ponds of Lake Guchenghu Basin, China: Occurrence, Temporal Variations, and Ecological Risks

Antibiotics are widely used in aquaculture, however, this often results in undesirable ecological effects. To evaluate the occurrence, temporal variations, and ecological risk of antibiotics in five crab ponds of Lake Guchenghu Basin, China, 44 antibiotics from nine classes were analyzed by rapid resolution liquid chromatography-tandem mass spectrometry (RRLC-MS/MS). Twelve antibiotics belonging to six classes were detected in the aqueous phase of five crab ponds, among which sulfonamides and macrolides were the predominant classes, and six compounds (sulfamonomethoxine, sulfadiazine, trimethoprim, erythromycin-H2O, monensin, and florfenicol) were frequently detected at high concentrations. In general, the antibiotic levels varied between different crab ponds, with the average concentrations ranging from 122 to 1440 ng/L. The antibiotic concentrations in crab ponds exhibited obvious seasonal variations, with the highest concentration and detection frequency detected in summer. Multivariate analysis showed that antibiotic concentrations were significantly correlated with environmental variables, such as total organic carbon, phosphate, ammonia nitrogen, and pH. Sulfadiazine, clarithromycin, erythromycin-H2O, and ciprofloxacin posed a high risk to algae, while the mixture of antibiotics could pose a high risk to aquatic organisms in the crab ponds. Overall, the usage of antibiotics in farming ponds should be comprehensively investigated and controlled to preserve a healthy aquaculture ecosystem.

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

Metabolomics of Hydrazine-Induced Hepatotoxicity in Rats for Discovering Potential Biomarkers.

Metabolic pathway disturbances associated with drug-induced liver injury remain unsatisfactorily characterized. Diagnostic biomarkers for hepatotoxicity have been used to minimize drug-induced liver injury and to increase the clinical safety. A metabolomics strategy using rapid-resolution liquid chromatography/tandem mass spectrometry (RRLC-MS/MS) analyses and multivariate statistics was implemented to identify potential biomarkers for hydrazine-induced hepatotoxicity. The global serum and urine metabolomics of 30 hydrazine-treated rats at 24 or 48 h postdosing and 24 healthy rats were characterized by a metabolomics approach. Multivariate statistical data analyses and receiver operating characteristic (ROC) curves were performed to identify the most significantly altered metabolites. The 16 most significant potential biomarkers were identified to be closely related to hydrazine-induced liver injury. The combination of these biomarkers had an area under the curve (AUC) > 0.85, with 100% specificity and sensitivity, respectively. This high-quality classification group included amino acids and their derivatives, glutathione metabolites, vitamins, fatty acids, intermediates of pyrimidine metabolism, and lipids. Additionally, metabolomics pathway analyses confirmed that phenylalanine, tyrosine, and tryptophan biosynthesis as well as tyrosine metabolism had great interactions with hydrazine-induced liver injury in rats. These discriminating metabolites might be useful in understanding the pathogenesis mechanisms of liver injury and provide good prospects for drug-induced liver injury diagnosis clinically.

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.

Pharmacokinetic Comparisons of Typical Constituents in Curcumae Rhizoma and Vinegar-Processed Curcumae Rhizoma after Oral Administration to Rats.

The Raw Curcumae Rhizoma (R-CR), included in the Chinese Pharmacopoeia Edition 2015, is a well-known Chinese herbal medicine. However, the vinegar-processed Curcumae Rhizoma (V-CR) is used more widely than R-CR. The pharmacokinetics comparison of R-CR and V-CR after oral administration to rats is poorly understood. A novel method, rapid resolution liquid chromatography-tandem mass spectrometry (RRLC-MS) coupled with a sensitive, specific, and convenient microdialysis sampling method, free from endogenous interference was developed in this research. The extracts of R-CR and V-CR were administered orally to each group of rats. The blood and liver microdialysis probes were positioned within the jugular vein toward the right atrium and the median lobe near the center of the liver, respectively. Then, a double-peak phenomenon was observed in the concentration-time curves of curdione in R-CR group, while it was not observed in V-CR group. The liver-to-blood distribution ratio of curdione in V-CR group increased significantly (P < 0.05) compared to that of R-CR group. However, compared with V-CR group, the pharmacokinetic parameters of curcumol exhibited no statistically significant differences from those of R-CR group. These results indicate that vinegar-processed procedure has influence on the pharmacokinetic process of Curcumae Rhizoma in/ns. RRLC-MS coupled with microdialysis system could be used to evaluate the pharmacokinetics of typical constituents in Curcumae Rhizoma after oral administration.

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.

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.

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.

Application of ionic liquid-based ultrasonic-assisted extraction followed by rapid resolution LC-ESI/MS to chromone analysis from radix saposhnikoviae

Ionic liquid-based ultrasonic-assisted extraction (ILUAE) was successfully applied to the extraction of the four chromones (prim-O-glucosylcimifugin, cimifugin, 5-O-methylvisammioside, and sec-O-glucosylhamaudol) from Saposhnikovia divaricata (Radix Saposhnikoviae) for the first time. A series of l-alkyl-3-methylimidazolium ILs differing in anion and cation compositions was evaluated for extraction efficiency, and [C3MIM]Br was selected as the optimal solvent. In addition, ultrasound extraction parameters were optimized, and the chromones were directly quantified and analyzed by rapid resolution liquid chromatography-electrospray ionization/mass spectrometry (RRLC-ESI/MS). The optimal conditions were as follows: 0.4 M concentration of [C3MIM]Br, 20:1 solvent to solid ratio, and ultrasonic time, temperature, and frequency of 5 min, 40 °C, and 50 kHz, respectively. This approach obtained the highest extraction yield of 10.188 ± 0.473 mg g−1 for total chromones. Compared with regular UAE, the proposed approach exhibited a higher efficiency (61.56% increase) and shorter extraction time (nine times shorter). Also, ILUAE was an efficient, rapid, and simple sample preparation technique for extraction of chromones, and the established RRLC-DAD method could serve as a rapid and effective technique for extracting chromones from Radix Saposhnikoviae.

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.

Effect of addition of overripe seeds from white grape by-products during red wine fermentation on wine colour and phenolic composition

The effect of the fermentative addition of overripe seeds by-product (Pedro Ximénez white grapes, 3 g/L) on the phenolic composition and colour of red wines was studied by rapid resolution liquid chromatography (RRLC-MS) and Differential Colorimetry. Overripe seeds summited to postharvest dehydration by sun drying directly demonstrated were rich in phenolics such as gallic acid, epicatechin, and procyanidin B2 3-O-gallate, and hence a source of copigments capable to stabilize wine anthocyanins. The fermentative addition of overripe seeds led to Syrah wines with significant higher content of anthocyanins and procyanidins than traditional macerated wines, which had a positive effect on colour quality and stability. With respect to the colour quality, visually perceptible colour differences were found respect to the traditional macerated wines (ΔE*ab > 3), getting overripe seeds macerated wines with bluish hues and higher chroma values. Moreover, Differential Colorimetry demonstrated that the addition of overripe seeds induces lower colour modification in wines during stabilization and, in consequence, higher colour stability.

Targeted Metabolomic Analysis of Polyphenols with Antioxidant Activity in Sour Guava (Psidium friedrichsthalianum Nied.) Fruit.

Psidium is a genus of tropical bushes belonging to the Myrtaceae family distributed in Central and South America. The polar extract of Psidium friedrichsthalianum Nied. was partitioned with ethyl ether, ethyl acetate, and n-butanol, and the total phenolic content and antioxidant activity were measured by Folin-Ciocalteu and ABTS assays, respectively. The ethyl acetate fraction exhibited both the highest phenolic content and antioxidant activity. Due to the complexity of this fraction, an analytical method for the comprehensive profiling of phenolic compounds was done by UPLC-ESI/QqQ in MRM (multiple reaction monitoring) mode. In this targeted analysis, 22 phenolic compounds were identified, among which several hydroxybenzoic, phenylacetic, and hydroxycinnamic acid derivatives were found. This is the first time that (+)-catechin, procyanidin B1, procyanidin B2, and (−)-epicatechin have been reported as constituents of sour guava. A fractionation by exclusion size, C18-column chromatography, and preparative RRLC (rapid resolution liquid chromatography) allowed us to confirm the presence of ellagic acid and isomeric procyanidins B, well-known bioactive compounds. The content of phenolic compounds in this fruit shows its potential for the development of functional foods.

Development of new efficient method for isolation of phenolics from sea algae prior to their rapid resolution liquid chromatographic–tandem mass spectrometric determination

The extraction of phenolic compounds from 4 different sea algae samples, three brown algae (Cystoseira abies-marina, C. abies-marina grinded under cryogenic conditions with liquid nitrogen, Undaria pinnatifida and Sargassum muticum) and one red algae (Chondrus crispus) via solid phase extraction using micro-elution solid-phase extraction (μ-SPE) plate method was studied. Prior to μ-SPE, 50mg of algae with 80% methanol mixture was extracted in hyphenated series by various extraction techniques, such as pressurized liquid extraction and Ika Ultra-Turrax® Tube Drive, in combination with ultrasound assisted extraction. The μ-SPE plate technique reduced the time of sample pre-treatment thanks to higher sensitivity and pre-concentration effect. Selected groups of benzoic acid derivatives (p-hydroxybenzoic, protocatechuic, gallic, vanillic, and syringic acids), hydroxybenzaldehydes (4-hydroxybenzaldehyde, and 3,4-dihydroxybenzaldehyde), and cinnamic acid derivatives (p-coumaric, caffeic, ferulic, sinapic, and chlorogenic acids) were determined using rapid resolution liquid chromatography coupled to mass spectrometry detection with negative ion electrospray ionization (RRLC-ESI–MS) using multiple reactions monitoring. LOQs of measured samples varied in the range 0.23–1.68ng/mL and LODs in the range 0.07–0.52ng/mL. The applied method allowed a simultaneous determination of phenolics (i.e. free, esters soluble in methanol, glycosides, and esters insoluble in methanol) in less than 5min (including alkaline or acidic hydrolysis of raw extracts) from sea algae extracts.