Following oral intake of Panax ginseng, major ginsenosides are metabolized to deglycosylated ginsenosides by gut microbiota before absorption into the blood. As the composition of gut microbiota varies between individuals, metabolic activities are significantly different. We selected 6 rats with low efficiency metabolism (LEM) and 6 rats with high efficiency metabolism (HEM) from 60 rats following oral administration of Panax ginseng extract, and analyzed their gut microbiota composition using Illumina HiSeq sequencing of the 16S rRNA gene. The components of gut microbiota between the LEM and HEM groups were significantly different. Between the 2 groups, S24-7, Alcaligenaceae, and Erysipelotrichaceae occupied most OTUs of the HEM group, which was notably higher than the LEM group. Furthermore, we isolated Bifidobacterium animalis GM1 that could convert the ginsenoside Rb1 to Rd. The result implies that these specific intestinal bacteria may dominate the metabolism of Panax ginseng.Keywords: ginsenoside; LC-MS/MS; metabolism; Panax ginseng; rats gut microbiota;

BackgroundIn general, after Panax ginseng is administered orally, intestinal microbes play a crucial role in its degradation and metabolization process. Studies on the metabolism of P. ginseng by microflora are important for obtaining a better understanding of their biological effects.MethodsIn vitro biotransformation of P. ginseng extract by rat intestinal microflora was investigated at 37°C for 24 h, and the simultaneous determination of the metabolites and metabolic profile of P. ginseng saponins by rat intestinal microflora was achieved using LC–MS/MS.ResultsA total of seven ginsenosides were detected in the P. ginseng extract, including ginsenosides Rg1, Re, Rf, Rb1, Rc, Rb2, and Rd. In the transformed P. ginseng samples, considerable amounts of deglycosylated metabolite compound K and Rh1 were detected. In addition, minimal amounts of deglycosylated metabolites (ginsenosides Rg2, F1, F2, Rg3, and protopanaxatriol-type ginsenosides) and untransformed ginsenosides Re, Rg1, and Rd were detected at 24 h. The results indicated that the primary metabolites are compound K and Rh1, and the protopanaxadiol-type ginsenosides were more easily metabolized than protopanaxatriol-type ginsenosides.ConclusionThis is the first report of the identification and quantification of the metabolism and metabolic profile of P. ginseng extract in rat intestinal microflora using LC–MS/MS. The current study provided new insights for studying the metabolism and active metabolites of P. ginseng.

To study the β-glucosidase gene (bgy1) from Lactobacillus brevis that was cloned and expressed in Escherichia coli BL21 (DE3) and then using it for the biotransformation of gypenoside XVII.The bgy1 gene consists of 2283 bp encoding 761 amino acids, with homology to the glycosyl hydrolase family-3 protein domain. The enzyme (Bgy1) hydrolyzed the glucose moieties at the C-3 position and the outer glucose moieties at the C-20 position of gypenoside XVII. Using 0.1 mg enzyme ml−1 in 20 mM sodium phosphate buffer at 30 °C and pH 6.0, 1 mg gypenoside XVII ml−1 was transformed into 0.58 mg compound K ml−1 within 6 h, with a corresponding molar conversion yield of 89 %.The recombinant Bgy1 is considered potentially useful for the practical preparation of compound K.

•We established a novel PAE analysis method.•The method can overcome the problems of the blank value in PAE analysis.•Contents and profiles of PAEs were analysed in 78 foodstuffs.•PAEs exposure was estimated from foodstuffs sold in the Chinese market.Phthalate esters (PAEs) are commonly used as nonreactive plasticisers in vinyl plastics to increase the flexibility of plastic polymers. Numerous studies have indicated that the PAEs as a class of endocrine-disrupting chemicals. In addition, the studies have also shown that a major source of human exposure to phthalates is the diet. To date, the largest problem in PAEs analysis is the high blank value because PAEs are widely used in various applications and products. To overcome this shortcoming, gas purge microsyringe extraction (GP-MSE) was applied, which established a new and low-blank-value analytical method for PAE analysis to analyse PAEs in foodstuffs. In this study, GP-MSE was used as a clean-up method, and the overall recoveries ranged from 85.7 to 102.6%, and the RSD was less than 10%. More importantly, this method can overcome the problem of the high blank value in PAE analysis. This method was applied for measuring PAEs in 78 foodstuffs. The results showed that a wide variety of PAE concentrations were found in the different groups, and the content of PAEs (varies from 658 to 1610 ng g−1 fresh weight) is greatest in seafood. The concentrations were in the following order: DEHP > DBP > DEP ≈ DMP > BBP ≈ DNOP. Finally, the daily intake of PAEs was estimated for adults based on the levels of PAEs in foodstuffs. The total EDIdiet values of 3.2 and 12.9 μg kg−1 bw d−1 were calculated for DEHP based on the mean and highest concentrations in foodstuffs, respectively.Low blank value analytical method for analysis of PAEs in the foodstuff.

•We developed a novel and rapid analysis method of OPPs in edible fungus.•OPPs were directly extracted from edible fungus using GP-MSE without cleanup step.•The process is completed within 2 min with good repeatability and recovery.•The extracts are directly injected into the GC–MS analysis.In this work a new analytical method for a rapid and simultaneous determination of 28 organophosphorus pesticides (OPPs) residues in edible fungus using gas purge microsyringe extraction (GP-MSE), coupled with on-line gas chromatography–mass spectrometry (GP-MSE-GC-MS) has been developed and optimized. GP-MSE, a novel gas flow liquid-phase microextraction technique, has been then fruitfully used as innovative and one-step extraction procedure, allowing a direct injection into the gas chromatograph coupled with a mass spectrometry detector (GC–MS) system without any further cleaning step. Once optimized, the GP-MSE-GC-MS analysis procedure showed reproducibility values, resolutions, linear responses, detection and quantification limits that allowed to consider this method suitable for the analysis of the 28 OPPs in real samples. Furthermore, OPP recoveries and the relative standard deviations (RSDs) ranged from 85.26% to 100.21%, and from 1.6% to 6.9%, respectively. This procedure was then used for the analysis of real samples and the obtained results were compared with those of ultrasonic extraction–solid phase extraction. Among the 28 OPPs, 14 of them were found in Lentinus edodes and Enoki mushrooms fungus samples, with a total concentrations of 112.7 and 210.7 μg kg−1, respectively. This work demonstrated then that GP-MSE-GC-MS provided a highly efficient, solvent-saving, accurate and sensitive quantitative analysis method for a rapid determination of OPPs in edible fungus.

•We developed an etched stainless steel wire/ionic liquid-solid phase microextraction.•No sample pre-treatment required.•Extraction parameters were examined in our study.•Comparable efficiency with commercial PA fibre for the analysis of APs in water samples.•This technique is efficient and convenient for extraction of APs from water sample.In this study, a stainless steel wire/ionic liquid-solid phase microextraction technique was developed for the direct extraction of APs from water samples. Some parameters were optimised, such as selection of the substrate and ILs, extraction time, extraction temperature, stirring rate and sample pH, etc. The experimental data demonstrated that the etched stainless steel wire was a suitable substrate for IL-coated SPME. The coating was prepared by directly depositing the ILs onto the surface of the etched stainless steel wire, which exhibited a porous structure and a high surface area. The [C8MIM][PF6] IL exhibited maximum efficiency with an extraction time of 30 min, and the aqueous sample was maintained at 40 °C and adjusted to pH 2 under stirring conditions. The enrichment factor of the IL coating for the four APs ranged from 1382 to 4779, the detection limits (LOD, S/N=3) of the four APs ranged from 0.01 to 0.04 ng mL−1 and the RSD values for purified water spiked with APs ranged from 4.0 to 11.8% (n=3). The calibration graphs were linear in the concentration range from 0.5 to 200 ng mL−1 (R2>0.9569). The optimised method was successfully applied for the analysis of real water samples, and the method was suitable for the extraction of APs from water samples.

•We establish the theoretical system of GP-MSE which is a novel microextraction.•The article elaborates on extraction kinetic and main parameters of GP-MSE.•Direct extraction of PAHs from plant samples is completed and fast (within 6 min).•The results provide a theoretical guide in the real application of GP-MSE.Gas purge-microsyringe extraction (GP-MSE) is a rapid and exhaustive microextraction technique for volatile and semivolatile compounds. In this study, a theoretical system of GP-MSE was established by directly extracting and analyzing 16 kinds of polycyclic aromatic hydrocarbons (PAHs) from plant samples. On the basis of theoretical consideration, a full factorial experimental design was first used to evaluate the main effects and interactions of the experimental parameters affecting the extraction efficiency. Further experiments were carried out to determine the extraction kinetics and desorption temperature-dependent. The results indicated that three factors, namely desorption temperature (temperature of sample phase) Td, extraction time t, and gas flow rate u, had a significantly positive effect on the extraction efficiency of GP-MSE for PAHs. Extraction processes of PAHs in plant samples followed by first-order kinetics (relative coefficient R2 of simulation curves were 0.731–1.000, with an average of 0.958 and 4.06% relative standard deviation), and obviously depended on the desorption temperature. Furthermore, the effect of the matrix was determined from the difference in Eapp,d. Finally, satisfactory recoveries of 16 PAHs were obtained using optimal parameters. The study demonstrated that GP-MSE could provide a rapid and exhaustive means of direct extraction of PAHs from plant samples. The extraction kinetics were similar that of the inverse process of the desorption kinetics of the sample phase.

•Overview summarizes of the application strategies of microextraction techniques in plant analysis.•Mainly summarizes microextraction methods including: SPME, SDME, HF-LPME, DLLME, SBSE, GP-MSE.•Target analytes mainly including plant secondary metabolites, VOCs and pesticides in this review.•Future trends of microextraction techniques in plant analysis were described.Vegetables and fruits are necessary for human health, and traditional Chinese medicine that uses plant materials can cure diseases. Thus, understanding the composition of plant matrix has gained increased attention in recent years. Since plant matrix is very complex, the extraction, separation and quantitation of these chemicals are challenging.In this review we focus on the microextraction techniques used in the determination of volatile and semivolatile organic compounds (such as esters, alcohols, aldehydes, hydrocarbons, ketones, terpenes, sesquiterpene, phenols, acids, plant secondary metabolites and pesticides) from plants (e.g., fruits, vegetables, medicinal plants, tree leaves, etc.). These microextraction techniques include: solid phase microextraction (SPME), stir-bar sorptive extraction (SBSE), single drop microextraction (SDME), hollow fiber liquid phase microextraction (HF-LPME), dispersive liquid liquid microextraction (DLLME), and gas purge microsyringe extraction (GP-MSE). We have taken into consideration papers published from 2008 to the end of January 2013, and provided critical and interpretative review on these techniques, and formulated future trends in microextraction for the determination of volatile and semivolatile compounds from plants.Microextraction technique in the determination of volatile and semivolatile organic compounds from plants.

The presence of perfluorocarboxylates (PFCAs) in the environment is of increasing concern due to their possible toxicity to humans and bioaccumulation in organisms. PFCAs are frequently found in river water, sediment and organisms and sometimes even in groundwater. In order to quantitatively determine these PFCAs, a fast derivatization coupled with a liquid chromatography–ultraviolet detector–electrospray ionization-tandem mass spectrometry (LC–UV–ESI-MS/MS) method was developed. The PFCAs were quantitatively converted to their corresponding phenacyl esters using p-bromophenacyl bromide as the derivatization reagent. Under optimized reaction conditions, the conversion yield of the PFCAs ranged from 86 to 92% with low %RSD. The typical derivatization product (p-bromophenacyl bromide perfluorooctanoate) was characterized by 1H NMR, 13C NMR, FT-IR and mass spectrometry. UPLC with a BEH C18 column and CAN/H2O (8/2, v/v) as a mobile phase were used to separate the derivatives. The analytes were completely eluted within 6 min and multidimensional detection using UV at 260 nm and ESI–MRM in the negative ion mode were carried out. Bromide isotopic characteristic fragment ions appeared in the first Q1 scans, and four daughter ions of the MRMs at m/z [M−H − 222]−, [M−H − 250]–, [M−H − 278]− and [M−H − 316]− were used for quantification and confirmation. The mass spectral information ensured accurate identification of the analytes even when the sample matrices were complex. The method successfully eliminated the PFCAs background problems originating from polymeric parts in liquid chromatographic systems. The LODs of the method were lower than 5 ng mL−1, and the relative standard deviation (RSD%) values ranged from 5.2 to 9.8%. The method was successfully applied for the quantification of PFCAs in river water contaminated by industrial wastewater, and this indicated that the method was useful in the determination of PFCAs in environmental samples.Highlights► PFCAs were quantitatively converted to their corresponding phenacyl esters. ► Conversion yield of the PFCAs determined ranged from 86.5 to 92%. ► Multidimensional liquid chromatography–UV–MS/MS analysis was possible. ► The method successfully eliminated the background noise from the LC system. ► This is a useful technique for the determination of PFCAs from complex samples.

Sample pretreatment before chromatographic analysis is the most time consuming and error prone part of analytical procedures, yet it is a key factor in the final success of the analysis. A quantitative and fast liquid phase microextraction technique termed as gas purge microsyringe extraction (GP-MSE) has been developed for simultaneous direct gas chromatography–mass spectrometry (GC–MS) analysis of volatile and semivolatile chemicals without cleanup process. Use of a gas flowing system, temperature control and a conventional microsyringe greatly increased the surface area of the liquid phase micro solvent, and led to quantitative recoveries of both volatile and semivolatile chemicals within short extraction time of only 2 min. Recoveries of polycyclic aromatic hydrocarbons (PAHs), organochlorine pesticides (OCPs) and alkylphenols (APs) determined were 85–107%, and reproducibility was between 2.8% and 8.5%. In particular, the technique shows high sensitivity for semivolatile chemicals which is difficult to achieve in other sample pretreatment techniques such as headspace-liquid phase microextraction. The variables affecting extraction efficiency such as gas flow rate, extraction time, extracting solvent type, temperature of sample and extracting solvent were investigated. Finally, the technique was evaluated to determine PAHs, APs and OCPs from plant and soil samples. The experimental results demonstrated that the technique is economic, sensitive to both volatile and semivolatile chemicals, is fast, simple to operate, and allows quantitative extraction. On-site monitoring of volatile and semivolatile chemicals is now possible using this technique due to the simplification and speed of sample treatment.

The gas purge microsyringe extraction (GP-MSE) technique offers quantitative and simultaneous extraction, and rapid gas chromatographic–mass spectrometric determination of volatile and semivolatile chemicals is possible. To simplify the application, a new automatic temperature control system was developed here. Stable heating and cooling over a wide range of temperatures were achieved using a micro-heater and thermoelectric cooler under varying gas flow conditions. Temperatures could be accurately controlled in the range 20–350 °C (heating) and 20 to −4 °C (cooling). Temperature effects on the extraction performance of the GP-MSE were experimentally investigated by comparing the recoveries of polycyclic aromatic hydrocarbons (PAHs) under various experimental conditions. A sample treatment was completed within 3 min, which is much less than the time required for chromatographic analysis. The recovery of chemicals determined ranged from 81 to 96%. High reproducibility data (RSD ≤ 5%) were obtained for direct extraction of various analytes in spiked complex plant and biological samples. The data show that the heating and cooling system has potential applications in GP-MSE system for the direct determination of various kinds of volatile and semivolatile chemicals from complex matrices without any, or only minor, sample pretreatment.Highlights► An automatic temperature control system was developed here. ► The wide ranges of stable heating and cooling temperatures were achieved using micro-heater and thermoelectric cooler, respectively. ► It was applied to “Gas purge microsyringe extraction (GP-MSE)” system. ► The quantitative extraction recoveries were obtained within 3 min.

The climatic characteristic is a major parameter affecting on the distribution variation of organic pollutants such as polycyclic aromatic hydrocarbons (PAHs). The Tumen River is located in Northeastern of China. The winter era lasts for more than 5 months in a year, and the river water was frozen and covered by ice phase. Coal combustion is an essential heating source in the Tumen River Basin. The objective of this research is to study ice phase effect on the seasonal variation of PAHs in the Tumen River environment.Samples were collected from 13 sites along the River in March, July, October, and December of 2008. In addition, the ice sample, under ice water and air particulate were also collected in winter. The samples were analyzed for 16 PAHs (naphthalene, acenaphthylene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, beazo[a]anthene, chrysene, beazo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, indeno(1,2,3-cd)pyrene, dibenz(a,h)anthracene, and benzo(ghi)perylene). The compounds were extracted from the water samples and solid samples using LLE and Soxhlet extraction technique, respectively, and it is determined by gas chromatography–mass spectrometry.Among 16 PAHs, fluorene, phenanthrene, and pyrene were found to be present in high concentrations and at high detection frequencies. The total concentration of PAHs in the water, particulate, sediment and ice phase ranged from 35.1–1.05 × 103 ng L−1, 25.4–817 ng L−1, 117–562 ng g−1and 62.8–136 ng g−1, respectively. The levels of PAHs were generally higher in spring than other seasons. The ice phase in winter acts like a major reservoir of the pollutants and it is major contributor on the seasonal variation of PAHs in Tumen River. The PAHs found in water, particulate, and sediment in the Tumen River were possibly derived from similar pollution sources a proposition based on the compositions and isomer ratios of PAHs.The distribution of PAHs was showed clear seasonal variation in the Tumen River environment, the ice phase and air pollution look like an important factor affecting on the seasonal variation.The ice phase as an important factor affecting on the seasonal variation of PAHs in Tumen River environment. Further studies regarding the effects of air pollution on the river and the mechanisms of migration and transformation of them in the environment are currently being conducted in our laboratory.

Mesoporous silica, SBA-15 was successfully functionalized with quinoline moiety, and was applied as a matrix in the MALDI-TOF-MS analysis of small molecules. The modified SBA-15 material [SBA-15-8-(3-(triethoxysilyl)propoxy) quinoline, SBA-15-8QSi] was obtained by using calcined SBA-15 and 8-hydroxy quinoline. The structure of the functionalized mesoporous material was systemically characterized by TEM, the N2 adsorption-desorption isotherm technique and FT-IR spectra. Compared with DHB and SBA-15, SBA-15-8QSi demonstrated several advantages in the analysis of small molecules with MALDI-TOF-MS, such as less background interference ions, high homogeneity, and better reproducibility. Based on these results, the various analytical parameters were optimized. The ideal operating conditions were (1) methanol used as the dissolving solvent; (2) sample first dropping method; (3) a ratio between the analyte and the matrix of 3.5:10. Under these optimization conditions, a low detection limit (8 pmol for L-Arginine-HCl) and high reproducibility (≤29%) were obtained. This technique was successfully applied to the analysis of various types of small molecules, such as saccharides, amino acids, metabolites, and natural honey.SBA-15 was functionalized with quinoline moiety, it is demonstrated less background, high homogeneity, and better reproducibility with low detection limit in the MALDI-TOF-MASS analysis.Figure optionsDownload full-size imageDownload high-quality image (88 K)Download as PowerPoint slide

Of the many parameters that affect the enrichment factors in headspace liquid-phase microextraction, in this study, we systematically investigated the influence of the vapor pressure of the extracting solvent. Seven extracting solvents with different vapor pressures were selected and tested. It was found that the vapor pressure of the extracting solvent dramatically affects the enrichment factor and the factor was increasing by decreasing the extracting solvent vapor pressure under given experimental conditions. The result was validated for volatile organic compounds such as polynuclear aromatic hydrocarbons, organochlorine pesticides and polychlorinated biphenyls.

A simple and economical cleanup technique was developed to determine alkylphenols by GC–MS from biological extracts containing relatively high lipids. The lipids were successfully removed from bivalve extracts through a two-step cleanup. The new method is a combination of Florisil adsorption chromatography and silyl derivatization technique. Low and high (non-polar and highly polar) molecular weight lipids were removed from the biota extract with deactivated Florisil column in the first step. And in the second step, middle molecular weight (middle polar) lipids were removed in an activated Florisil column after the alkylphenols were converted to corresponding silyl derivatives with bis(trimethylsilyl)trifluoroacetamide (BSTFA). On the basis of the above results, a simple cleanup kit was developed for convenience. The technique was optimized with reference to the activity of packing materials and polarity of eluting solvents. Only 3 g of Florisil, 25 mL of hexane and 10 mL of dichloromethane were required for one sample. The recoveries of alkylphenols from spiked samples varied from 88 to 103% with a low relative standard deviation (mean value: 5.3%) and the recovery was similar or even higher than other methods currently in use. The technique was successfully applied to mussel samples from Masan Bay, South Korea. Simultaneous measurement of these compounds in water, sediment and biota; the resulting bio-concentration factor and their relationships confirm previously published works, validating the method applied.