Simultaneous determination of cyanide and thiocyanate in milk by GC-MS/MS using cetyltrimethylammonium bromide as both phase transfer catalyst and protein precipitant.

A method was developed for simultaneous determination of cyanide and thiocyanate in milk by gas chromatography-tandem quadrupole mass spectrometry (GC-MS/MS). Cyanide and thiocyanate were derivatized with pentafluorobenzyl bromide (PFBBr) as PFB-CN and PFB-SCN, respectively. Cetyltrimethylammonium bromide (CTAB) was employed both as a phase transfer catalyst and a protein precipitant in the sample pretreatment, which facilitates the separation of the organic and aqueous phases, and greatly simplifies the pretreatment procedures to achieve simultaneous and rapid determination of cyanide and thiocyanate. Under the optimized conditions, the limits of detection (LODs) of cyanide and thiocyanate in milk were 0.006 mg/kg and 0.015 mg/kg, and the spiked recoveries ranged from 90.1% to 98.2% and from 91.8% to 98.9% with relative standard deviations (RSDs) less than 18.9% and 15.2%, respectively. The proposed method was validated as a simple, fast and highly sensitive method for the determination of cyanide and thiocyanate in milk.

The relationship between plasma free fatty acids, cognitive function and structural integrity of the brain in middle-aged healthy humans.

BACKGROUND: The cerebral composition of ω-3 and ω-6 polyunsaturated fatty acids (PUFAs) is believed to influence cognitive function and structural damage of the aging brain. However, existing data is inconsistent. MATERIALS AND METHODS: This retrospective study explored the association between free plasma PUFA concentrations, cognitive function and brain structure atrophy in a well-characterized community-dwelling cohort of elderly individuals without stroke and dementia. Ten different fatty acids were analyzed in stored plasma samples from 391 non-demented elderly individuals by gas chromatography mass spectrometry. Neuropsychiatric tests capturing memory, executive function and visuopractical skills were performed in all participants. Brain atrophy was assessed by MRI in a subset of 167 individuals. RESULTS: Higher plasma concentrations of free ω-6 PUFAs (p = 0.042), and, in particular, linoleic acid (p = 0.01), were significantly associated with lower executive function. No significant association existed between ω-3 PUFA concentrations and cognitive functioning. The volume of the frontal lobes was inversely associated with ω-6 PUFAs, whereas ω-3 PUFAs were positively related with temporal lobe volumes. All associations did not withstand correction for multiple comparisons. CONCLUSIONS: Our study suggests subtle effects of PUFA imbalances on cognition and brain structure. Yet the observed associations are weak and unlikely to be of clinical relevance. The brain regions that seem to be most sensitive to imbalances of ω-3 and ω-6 PUFAs are the frontal and temporal lobes.

Determination of glyphosate, glufosinate and their major metabolites in urine by the UPLC-MS/MS method applicable to biomonitoring and epidemiological studies.

The preoccupation concerning glyphosate (GLYP) has rapidly grown over recent years, and the availability of genetically modified crops that are resistant to GLYP or glufosinate (GLUF) has increased the use of these herbicides. The debate surrounding the carcinogenicity of GLYP has raised interest and the desire to gain information on the level of exposure of the population. GLYP and aminomethylphosphonic acid (AMPA) are commonly simultaneously analysed. GLUF is sometimes also monitored, but its major metabolite, 3-[hydroxy(methyl)phosphinoyl]propionic acid (3MPPA), is rarely present in the method. Using a pentafluorobenzyl derivative to extract the analytes from human urine, we present a method that contains four important analytes to monitor human exposure to GLYP and GLUF. The use of the flash freeze technique speeds up the extraction process and requires less organic solvent than conventional liquid-liquid extraction. The limits of detection in the low µg/L range enable the use of this method for epidemiological studies. The results obtained for 35 volunteers from the Quebec City area are presented with the results from multiple interlaboratory comparisons (G-EQUAS, HBM4EU and OSEQAS). This methodology is currently being used in the Maternal-Infant Research on Environmental Chemicals (MIREC-ENDO) study and in the Canadian Health Measures Survey (CHMS).

Simultaneous quantification of straight-chain and branched-chain short chain fatty acids by gas chromatography mass spectrometry.

Biomedical research in areas such as metabolic disorders, neuromodulatory, and immunomodulatory conditions involves lipid metabolism and demands a reliable and inexpensive method for quantification of short chain fatty acids (SCFAs). We report a GC-MS method for analysis of all straight-chain and branched-chain SCFAs using pentafluorobenzyl bromide (PFBBr) as derivatization reagent. We optimized the derivatization and GC-MS conditions using a mixture containing all eight SCFA standards, i.e., five straight-chain and three branched-chain SCFAs. The optimal derivatization conditions were derivatization time 90 min, temperature 60 °C, pH 7, and (CH3)2CO:H2O ratio 2:1 (v:v). Comparing the performance of different GC column configurations, a 30 m DB-225ms hyphenated with a 30 m DB-5ms column in tandem showed the best separation of SCFAs. Using the optimized experiment conditions, we simultaneously detected all SCFAs with much improved detection limit, 0.244-0.977 µM. We further applied the developed method to measure the SCFAs in mouse feces and all SCFAs were successfully quantified. The recovery rates of the eight SCFAs ranged from 55.7% to 97.9%.

GC-MS Analysis of Short-Chain Fatty Acids in Feces, Cecum Content, and Blood Samples.

Short-chain fatty acids, the end products of fermentation of dietary fibers by the gut microbiota, have been shown to exert multiple effects on mammalian metabolism. For the analysis of short-chain fatty acids, gas chromatography-mass spectrometry is a very powerful and reliable method. Here, we describe a fast, reliable, and reproducible method for the separation and quantification of short-chain fatty acids in mouse feces, cecum content, and blood samples (i.e., plasma or serum) using gas chromatography-mass spectrometry. The short-chain fatty acids analyzed include acetic acid, propionic acid, butyric acid, valeric acid, hexanoic acid, and heptanoic acid.

GC-MS Analysis of Medium- and Long-Chain Fatty Acids in Blood Samples.

Our body contains a wide variety of fatty acids that differ in chain length, the degree of unsaturation, and location of the double bonds. As the various fatty acids play distinct roles in health and disease, methods that can specifically determine the fatty acid profile are needed for fundamental and clinical studies. Here we describe a method for the separation and quantification of fatty acids ranging from 8 to 24 carbon chain lengths in blood samples using gas chromatography-mass spectrometry following derivatization using pentafluorobenzyl bromide. This method quantitatively monitors fatty acid composition in a manner that satisfies the requirements for comprehensiveness, sensitivity, and accuracy.

Detecting human urinary thiocyanate by gas chromatography with pre-column derivatization / 中国职业医学

OBJECTIVE: To establish a method for detecting human urinary thiocyanate by gas chromatographic and pre-column derivatization with 2,3,4,5,6-pentafluorobenzyl bromide( PFB-Br). METHODS: A total of 20. 0 μL of urine was taken and 1. 0 m L of acetonitrile and 100. 0 μL of PFB-Br were added for derivative reaction. The gas chromatography was directly used for measurement. RESULTS: The urinary thiocyanate concentration showed a good linear range of 1. 000-10. 000 mg/L. The linear correlation coefficient was 0. 999 6. The minimum detection concentration was 0. 112 mg/L,and the minimum quantitative concentration was 0. 411 mg/L( 20. 0 μL urine sample). The standard recovery rate was 97. 22%-102. 04%.The within-run relative standard deviation( RSD) of this method was 1. 56%-5. 35%. The between-run RSD was 1. 46%-5. 10%. Hydrocyanic acid ions interfered with the measurement. Other common inorganic ions such as chloride,sulfate,and nitrate ions did not interfere with the measurement results. The samples can be stored at 4 ℃ for at least 15 days. CONCLUSION: This method is suitable for detecting human urinary thiocyanate.

Analysis of naphthenic acid mixtures as pentafluorobenzyl derivatives by gas chromatography-electron impact mass spectrometry.

In this study, we report for the first time the efficiency of pentafluorobenzyl bromide (PFBBr) for naphthenic acid (NA) mixtures derivatization, and the comparison in the optimal conditions to the most common NAs derivatization reagents, BF3/MeOH and N-(t-butyldimethylsilyl)-N-methyltrifluoroacetamide (MTBSTFA). Naphthenic acids are carboxylic acid mixtures of petrochemical origin. These compounds are important for the oil industry because of their corrosive properties, which can damage oil distillation infrastructure. Moreover, NAs are commercially used in a wide range of products such as paint and ink driers, wood and fabric preservatives, fuel additives, emulsifiers, and surfactants. Naphthenic acids have also been found in sediments after major oils spills in the United States and South Korea. Furthermore, the toxicity of the oil sands process-affected water (OSPW), product of the oil sands extraction activities in Canada's oil sands, has largely been attributed to NAs. One of the main challenges for the chromatographic analysis of these mixtures is the resolution of the components. The derivatization optimization was achieved using surface response analysis with molar ratio and time as factors for derivatization signal yield. After gas chromatography-electron impact mass spectrometry (GC/EIMS) analysis of a mixture of NA standards, it was found that the signal produced by PFB-derivatives was 2.3 and 1.4 times higher than the signal produced by methylated and MTBS-derivatives, respectively. The pentafluorobenzyl derivatives have a characteristic fragment ion at 181m/z that is diagnostic for the differentiation of carboxylic and non-carboxylic acid components within mixtures. In the analysis of a Sigma and a Merichem derivatized oil extract NA mixtures, it was found that some peaks lack the characteristic fragment ion; therefore they are not carboxylic acids. Open column chromatography was used to obtain a hexane and a methanol fraction of the Sigma and Merichem mixtures. The components in the hexane fraction, presumably hydrocarbons that did not react with PFBBr were ~7% by weight. The effectiveness of PFBBr was confirmed when the two NA oil extracts were spiked with 8 distinct NA standards and identified by GC/EIMS in the methanol fraction. Here, we also report retention indices of the methyl, MTBS and PFB derivatives of these 8 NAs. The use of PFBBr increases sensitivity, chromatographic resolution, and identification accuracy for the analysis of standards and mixtures of NAs compared to MTBSTFA and BF3/MeOH. This methodology will have wide applications in the elucidation of NA mixtures.

Critical practical aspects in the application of liquid chromatography-mass spectrometric studies for the characterization of impurities and degradation products.

Liquid chromatography-mass spectrometry (LC-MS) is considered today as a mainstay tool for the structure characterization of minor components like impurities (IMPs) and degradation products (DPs) in drug substances and products. A multi-step systematic strategy for the purpose involves high resolution mass and multi-stage mass studies on both the drug and IMPs/DPs, followed by comparison of their fragmentation profiles. Its successful application requires consideration of many practical aspects at each step. The same are critically discussed in this review.