A new method to analysis synthetic acetic acid to vinegar: Hydrogen isotope ratio at the methyl site of acetic acid in vinegar by GC-IRMS.

Acetic acid is the key organic substance used to verify the authenticity of vinegar. A new method for precisely determining acetic acid δDCH3 in vinegar via gas chromatography -pyrolytic-stable isotope ratio mass spectrometry (GC-P-IRMS) was established. The δDCH3 values were obtained via calibration with a series of standards. The optimised method demonstrated a repeatability standard deviation within 3 ‰. The standard deviation of accuracy of the new method compared with that of the SNIF-NMR method was within 2.6 ‰. The synthetic acetic acid δDCH3 values was -136.7 ‰ ± 29.6 ‰, and the δDCH3 value of acetic acid in vinegar was -414.9 ‰ ± 40.5 ‰, with significant isotopic distribution characteristics. This methodology serves as a supplementary method for measuring the δDCH3 value of acetic acid in vinegar. It has advantages over other methods in terms of time, sensitivity and operability. And provides a new idea for solving the problem of analyzing substances in the presence of exchangeable groups.

A rapid method for the determination of stable hydrogen isotope ratios of acetic acid in vinegar.

RATIONALE: Vinegar is an everyday condiment made from fermented grains or fruits. It contains acetic acid which is the main organic material produced by fermentation. Vinegar suffers from the authenticity problem of exogenous adulteration due to the indistinguishability of low-cost chemical sources of synthetic acetic acid from acetic acid produced by fermentation. It is necessary to establish a simple and rapid measurement technique. METHODS: Determination was according to the total acid content of vinegar diluted with acetone to a certain concentration. Online separation and determination of acetic acid δD in vinegar were carried out using gas chromatography-pyrolysis-isotope ratio mass spectrometry. RESULTS: An HP-Plot/U column was selected for online separation of acetic acid and water with molecular sieve characteristics. At the same time, combined with the instrument blowback function to remove water. Dilute solvent acetone was treated with a molecular sieve to remove trace water. The reproducibility of this method is less than 3‰. The long-term stability is within a reasonable error range. The accuracy correlation coefficient is greater than 0.99. The δD values of acetic acid in vinegar (-264.5 ± 20.3‰) and from chemical sources (-30.5 ± 90.8‰) were obtained. CONCLUSIONS: A rapid method was developed for identification of different sources of acetic acid. These different sources of acetic acid exhibited significant hydrogen isotope distribution characteristics. Additionally, it was observed that the carboxyl hydrogen of acetic acid exhibited facile exchange with water. In future investigations, we aim to mitigate this interference.

Stable isotope ratio analysis of carbon to distinguish sialic acid from freshly stewed bird's nest products.

Freshly stewed bird's nest products are easily adulterated with exogenous synthetic sialic acid to enhance the grade of the products and sell at high prices. This paper identifies the carbon stable isotope characteristics of sialic acid from natural and commercially synthetic sources using stable isotope ratio mass spectrometry (IRMS). Specifically, an off-line pretreatment technique combined with on-line LC-IRMS was developed to accurately determine δ13C values of sialic acid in a freshly stewed bird's nest. This method has no obvious isotope fractionation and good reproducibility. EA-IRMS was used to determine the δ13C values of commercial sialic acid. The results showed that the δ13C values of sialic acid from natural and synthetic sources were -29.90% ± 0.42% and -16.26% ± 3.91%, respectively, with distinct carbon stable isotope distribution characteristics. By defining a δ13C threshold value of -28.54% for natural SA, additional commercial SA from a minimum of 10% can be identified. Therefore, δ13C was proposed as a suitable tool for verifying the authenticity of fresh stewed bird's nests on the market.

Mineralogical and elemental data for soil discriminating and geolocation tracing.

One of the key tasks of soil analysis in forensic sciences is to provide information about its diversities and geolocation. In fact, soil analysis is relevant for forensic geologists. In this study, a total of 80 soil samples were collected from eight Chinese cities (10 samples per city). Different minerals and their relative percentages were analyzed by the X-ray diffraction (XRD) method. In addition, the relative amounts of montmorillonite, kaolinite, amphibole, feldspar, calcite, and dolomite provided information about the origin of a soil, either if it came from a northern or southern city of China. The oxide weight percentages of 10 elements of Al2O3, SiO2, Fe2O3, K2O, Na2O, MgO, CaO, P2O5, MnO, and TiO2 were also obtained by using X-ray fluorescence (XRF) from the 80 soil samples. Moreover, principal component analysis (PCA) and hierarchical clustering analysis (HCA) methods were performed for dimensionality reduction, elemental marker identification and soils classification to the city they came from purposes. The eighty soils analyzed in this study could be tracked correctly to their city of origin. The K-Nearest Neighbors (KNN) model was done to evaluate the prediction ability based on the soil elemental composition, and it was confirmed by cross validation methods. The results demonstrated that mineralogical and elemental composition can provide powerful information for soil discrimination and source tracing.

What's in a wine? - A spot check of the integrity of European wine sold in China based on anthocyanin composition, stable isotope and glycerol impurity analysis.

The international wine market has been repeatedly hit by cases of fraud in recent decades. While several studies attested a special vulnerability of the fast growing wine business in China, reports on chemical analyses of commercial wine samples are rare. We examined 50 predominantly red wines with European labelling, which were purchased on the Chinese market, for fraud-relevant parameters. More than 20% of the tested samples revealed anomalies in relation to the stable isotope ratios of D/H, 18O/16O and 13C/12C, contents of technical glycerol by-products or anthocyanin composition. These results strongly suggested watering of the wines, chaptalisation, glycerol addition or the use of non-Vitis anthocyanin sources, respectively. Some of these samples also showed suspicious spelling errors or other irregularities in the labelling, but the majority appeared genuine to the eye. Hence, this spot check demonstrates the importance of chemical authenticity analysis of market samples in order to detect fraudulent products. Moreover, we used the same sample set for an evaluation of the Chinese standard method for carbon stable isotope determination of wine ethanol in comparison to the current OIV (International Organisation of Vine and Wine) standard method. The results of a Bland-Altman analysis indicated that the methods can be applied interchangeably. As the two methods differ in their workflow and in the requested equipment, this might eventually enable more laboratories to perform 13C/12C analysis of wine and spirits.

Vintage analysis of Chinese Baijiu by GC and 1H NMR combined with multivariable analysis.

Economical-driven counterfeit and inferior aged Chinese Baijiu has caused serious concern of publicity in China. In this study, a total of 167 authentic Chinese Baijiu samples with different vintages including 3 flavor types were carefully collected. Gas chromatography (GC) was used to determine main volatile components and proton nuclear magnetic resonance (1H NMR) spectroscopy was employed to obtain non-targeted fingerprints of Chinese Baijiu samples. Partial least squares regression (PLSR) models, which were confirmed by internal and external validation, were established for effectively identifying actual storage vintage of Chinese Baijiu with various brands, flavor types. Centering (Ctr), pareto scaling (Par), unit variance scaling (UV) data pretreatment methods, principal components (PCs), and three modified variable selection methods were proposed to successfully optimize the vintage model and effectively extract important vintage characteristic factors. This study demonstrated that NMR and GC combined with multivariate statistical analysis are effective tools for validating vintage authenticity of Chinese Baijiu.

Correlation between bacterial community succession and propionic acid during gray sufu fermentation.

In order to explore the correlation between the production of propionic acid (PA) and the succession of bacterial community during the fermentation of gray sufu, high-throughput sequencing and HPLC (High Performance Liquid Chromatography) were used to monitor the changes of bacterial community and metabolite content. The abundance and metabolite concentration of Propionibacterium increased rapidly in the early stage of fermentation. In the middle stage, the abundance of Lactobacillus began to increase, while the pH decreased rapidly. In the late stage, the concentration of PA began to decrease, but it remained at a high level at the end of fermentation. Correlation analysis showed that Lactobacillus and Bacillus had a strong negative correlation with PA and its precursor. The results showed that Fusobacterium, Providencia, Lactobacillus and Bacillus could be the key factors to reduce the PA content. This study provides a new idea for the quality control of traditional fermented food.

Survey of mineral oil hydrocarbons in infant formula from the Chinese market.

Mineral oil hydrocarbon (MOH) contamination of various foods in the past few decades has raised much concern due to its potential adverse health effects. Since infant formulas (IF) is the major food source for infants, it is necessary to understand MOH contamination level in IF and consequent potential food safety risks. Data on the contamination of IF by mineral oil are lacking in China. On the other hand, the analysis of MOH in food is difficult. There is no harmonised standard analytical method for testing MOHs in IF. GC-FID/MS was chosen as the analytical tool being more convenient for surveys at a national level. Fifty-one IFs comprising dairy milk-based IFs (n = 39) and goat milk-based IFs (n = 12), including different stages (Stage 1, 2 and 3), package type (metal cans and paper boxes) were collected in China market in 2018 for this survey. 17 of 51 IFs were found positive, but trace levels MOAH were found (≤0.7 mg/kg). For the positive samples, all the MOSH and MOAH hump fell into the C16-C25 fraction. MOH humps were found in all the 12 goat milk-based IFs, even 4 samples are reported with quantifiable values which are higher than the method defined LOQ. The highest quantifiable MOH contamination level of goat milk-based IFs were MOSH = 3.5 mg/kg and MOAH = 0.7 mg/kg. Further root cause analysis of contamination is highly recommended to control the MOH contamination for goat milk-based IFs.

Microbial Community Analyses Associated with Nine Varieties of Wine Grape Carposphere Based on High-Throughput Sequencing.

Understanding the composition of microbials on the grape carposphere may provide direct guidance for the wine brewing. In this work, 16S rRNA and ITS (Internal Transcribed Spacer) fungal amplicon sequencing were performed to investigate the differences of epiphytic microbial communities inhabiting different varieties of wine grape berries. The results showed that the dominated phylum of different wine grape carpospheres were Proteobacteria, Actinomycetes, Firmicutes, Gemmatimonadete, and Bacteroidetes. The dominant bacterial genera of different wine grape varieties were Pseudomonas, Arthrobacter, Bacillus, Pantoea, Planomicrobium, Massilia, Curtobacterium, Corynebacterium, Cellulomonas, Sphingomonas, and Microvirga. The fungal communities of the grapes were dominated by Ascomycota for all nine wine varieties. The dominant fungal genera on grape carposphere were Alternaria, Cladosporium, unclassified Capnodiales, Phoma, Rhodotorula, Cryptococcus, Aureobasidium, and Epicoccum. Community structure exerts a significant impact on bacterial Bray-Curtis dissimilarity on six red grapes and also a significant bacterial impact on three white grapes. Community structure exerts a significant impact on fungal Bray-Curtis dissimilarity on six red grapes but weak or no fungal impact on three white grapes. The results revealed that grape variety plays a significant role in shaping bacterial and fungal community, varieties can be distinguished based on the abundance of several key bacterial and fungal taxa.

Elemental profile and oxygen isotope ratio (δ18O) for verifying the geographical origin of Chinese wines.

The elemental profile and oxygen isotope ratio (δ18O) of 188 wine samples collected from the Changji, Mile, and Changli regions in China were analyzed by inductively coupled plasma mass spectrometry (ICP-MS), inductively coupled plasma optical emission spectroscopy (ICP-OES) and isotope ratio mass spectrometry (IRMS), respectively. By combining the data of δ18O and the concentration data of 52 elements, the analysis of variance (ANOVA) technique was firstly applied to obtain the important descriptors for the discrimination of the three geographical origins. Ca, Al, Mg, B, Fe, K, Rb, Mn, Na, P, Co, Ga, As, Sr, and δ18O were identified as the key explanatory factors. In the second step, the key elements were employed as input variables for the subsequent partial least squares discrimination analysis (PLS-DA) and support vector machine (SVM) analyses. Then, cross validation and random data splitting (training set: test set = 70:30, %) were performed to avoid the over-fitting problem. The average correct classification rates of the PLS-DA and SVM models for the training set were both 98%, while for the test set, these values were 95%, 97%, respectively. Thus, it was suggested that the combination of oxygen isotope ratio (δ18O) and elemental profile with multi-step multivariate analysis is a promising approach for the verification of the considered three geographical origins of Chinese wines.

Correction to: Measurement of the 15N/14N ratio of phenylalanine in fermentation matrix by isotope ratio mass spectrometry.

The publisher was alerted that the following important entry in the references of this article was missing.

Measurement of the 15N/14N ratio of phenylalanine in fermentation matrix by isotope ratio mass spectrometry.

OBJECTIVES: To determine the origin of 15N-labeled phenylalanine in microbial metabolic flux analysis using 15N as a tracer, a method for measuring phenylalanine δ15N using HPLC coupled with elemental analysis-isotope ratio mass spectrometry (EA-IRMS) was developed. RESULTS: The original source of the 15N-labeled phenylalanine was determined using this new method that consists of three steps: optimization of the HPLC conditions, evaluation of the isotope fractionation effects, and evaluation of the effect of pre-processing on the phenylalanine nitrogen stable isotope. In addition, the use of a 15N-labeled inorganic nitrogen source, rather than 15N-labeled amino acids, was explored using this method. CONCLUSIONS: The method described here can also be applied to the analysis of metabolic flux.

Rapid Method for the Determination of the Stable Oxygen Isotope Ratio of Water in Alcoholic Beverages.

This paper demonstrates the first successful application of an online pyrolysis technique for the direct determination of oxygen isotope ratios (δ(18)O) of water in alcoholic beverages. Similar water concentrations in each sample were achieved by adjustment with absolute ethyl alcohol, and then a fixed GC split ratio can be used. All of the organic ingredients were successfully separated from the analyte on a CP-PoraBond Q column and subsequently vented out, whereas water molecules were transferred into the reaction furnace and converted to CO. With the system presented, 15-30 µL of raw sample was diluted and can be analyzed repeatedly; the analytical precision was better than 0.4‰ (n = 5) in all cases, and more than 50 injections can be made per day. No apparent memory effect was observed even if water samples were injected using the same syringe; a strong correlation (R(2) = 0.9998) was found between the water δ(18)O of measured sample and that of working standards. There was no significant difference (p > 0.05) between the mean δ(18)O value and that obtained by the traditional method (CO2-water equilibration/isotope ratio mass spectrometry) and the newly developed method in this study. The advantages of this new method are its rapidity and straightforwardness, and less test portion is required.

[Determination of ethyl carbamate in wine by HPLC-FLD].

OBJECTIVE: To evaluate the method for determining ethyl carbamate (EC) in wine by HPLC-FLD. METHODS: Samples were derived and determined by HPLC with a fluorescence detector. Using a Agilent TC-C18 (250 mm x 4.6 mm x 5 microm) column and a mixture of CH3CN-0.02 mol/L CH3COONa as mobile phase for gradient elution. The incidence light wavelength was 233 nm, emission wavelength was 600 nm. The fluorescence reaction time was 30 min. The contents were calculated with an external standard. RESULTS: The linearity was good in the ranges of 10-500 microg/L for EC. The average recovery rates of EC was 93.7%-106.0%. The RSD was 1.4%-2.8%, and the limit of quantification was 15 microg/L. GC-MS and HPLC-FLD were compared to evaluate for determining EC content in wine. CONCLUSION: The HPLC-FLD method was simple, accurate and suitable for the determination of EC in multitudinous wine.

[Determination of 2,4,6-trichloroanisole in wine by head-space solid phase micro-extraction and gas chromatography-mass spectrometry].

OBJECTIVE: To establish a method of combining gas chromatography-mass spectrometry (GC-MS) with head-space solid phase micro-extraction (HS-SPME) for the detection of 2,4,6-trichloroanisole (2,4,6-TCA) in wine. METHODS: 2,4,6-TCA in wine were extracted by polydimethylsiloxane (PDMS,100 microm) fiber for 15 min under 35 degrees C in a water bath. The fiber was then transferred into the injection port of the GC-MS and separated by a CP-SIL 8CB-MS capillary column (30m x 0.25mm x 0.25 microm), and detected by mass spectrometry using d5-2,4,6-TCA as an internal standard. RESULTS: The linear range for 2,4,6-TCA was 5 (60ng/L, the detection limit was 0.06ng/L. The recovery of standard addition (5, 10, 20ng/L) was in the range of 79.8% (101.6% with relative standard deviations (RSD, n=5) < or = 7%. CONCLUSION: Organic solvents were avoided and the selection was sensitive for the method, which is suitable for the determination of 2,4,6-TCA in wine.

Methylotrophic methanogenesis governs the biogenic coal bed methane formation in Eastern Ordos Basin, China.

To identify themethanogenic pathways present in a deep coal bed methane (CBM) reservoir associated with Eastern Ordos Basin in China, a series of geochemical and microbiological studies was performed using gas and water samples produced from the Liulin CBM reservoir. The composition and stable isotopic ratios of CBM implied a mixed biogenic and thermogenic origin of the methane. Archaeal 16S rRNA gene analysis revealed the dominance of the methylotrophic methanogen Methanolobus in the water produced. The high potential of methane production by methylotrophic methanogens was found in the enrichments using the water samples amended with methanol and incubated at 25 and 35 °C. Methylotrophic methanogens were the dominant archaea in both enrichments as shown by polymerase chain reaction (PCR)­denaturing gradient gel electrophoresis (DGGE). Bacterial 16S rRNA gene analysis revealed that fermentative, sulfate-reducing, and nitrate-reducing bacteria inhabiting the water produced were a factor in coal biodegradation to fuel methanogens. These results suggested that past and ongoing biodegradation of coal by methylotrophic methanogens and syntrophic bacteria, as well as thermogenic CBM production, contributed to the Liulin CBM reserves associated with the Eastern Ordos Basin.