Interaction Chromatography-tandem Mass Spectrometry Research Articles

Development of a Cation Exchange SPE-HILIC-MS/MS Method for the Determination of Ningnanmycin Residues in Tea and Chrysanthemum.

Ningnanmycin is a widely used antibiotic in agricultural production that effectively controls fungal and viral diseases in tea trees and chrysanthemums. The polarity characteristic of ningnanmycin has posed limitations on the development of robust detection methods, thereby hindering effective monitoring and control measures. By combining cation exchange solid phase extraction (SPE) with hydrophilic interaction chromatography tandem mass spectrometry (HILIC-MS/MS), we have effectively tackled the issue pertaining to the separation and retention of ningnanmycin. The average recoveries of ningnanmycin in green tea, black tea, and chrysanthemum were 77.3-82.0%, 80.1-81.5%, and 74.0-80.0%, respectively. The intraday and interday relative standard deviations (RSDs) were below and equal to 7.7%. Good linearity was observed in the concentration range of 1-1000 μg/L (R2 > 0.998). The limits of detection (LODs) ranged from 1.1 μg/kg to 7.1 μg/kg, and the limits of quantification (LOQs) ranged from 3.6 μg/kg to 23.7 μg/kg for ningnanmycin. These results indicate the good accuracy, repeatability, reproducibility, and sensitivity of the method. It is suitable for detecting ningnanmycin in tea and chrysanthemum.

Simultaneous determination of 13 paralytic shellfish toxins and tetrodotoxin in marine shellfish by porous graphitic carbon solid-phase extraction and hydrophilic interaction chromatography-tandem mass spectrometry

The lethal neurotoxins, paralytic shellfish toxins (PSTs), and tetrodotoxin (TTX) have recently been found in marine shellfish from many coastal states. Herein, we applied a sensitive and reliable ultra-performance hydrophilic interaction chromatography (HILIC)-tandem mass spectrometry (MS) method to determine 13 PSTs and TTX in marine shellfish using a porous carbon solid-phase extraction (SPE). This in-house validation study required the development of a novel chromatographic separation using a HILIC-Z column, which was necessary to retain highly polar compounds. Using acetonitrile as the organic phase and ammonium formate-formic acid buffer as the aqueous phase, the quantitative analysis was carried out with an external standard method in the multiple reaction monitoring modes using positive electrospray ionization. To reduce interference, 1% aqueous acetic acid extracts of the shellfish samples were cleaned up by ion-pair SPE using a porous graphitic carbon cartridge. The calibration curves for PSTs and TTX were linear (R 2 > 0.995), and the sensitivity was good, with limits of detection (LODs) of 1.7–13.7 µg/kg, and limits of quantitation (LOQs) of 5.2–41.0 µg/kg. The recoveries were 76.5–95.5% with RSDs of 3.1–12.0%. Finally, We applied the method for the determination of PSTs and TTX in three batches of Nassarius showing excellent method accuracy against expected values.

Determination of caffeine, paraxanthine, theophylline and theobromine in premature infants by HILIC-MS/MS

Aim: Caffeine is a central nervous system stimulant, used to treat apnea of prematurity. A hydrophilic interaction chromatography-tandem mass spectrometry(HILIC-MS/MS) approach was developed to detect caffeine, paraxanthine, theophyllineand theobromine in premature infants. Method: Protein precipitation of plasma samples (10μl) was carried out by treating with acetonitrile containing caffeine-13C3. The separation was carried out on an ACQUITY HPLC® BEH HILIC column. Caffeine and its metabolites were quantified by multiple reaction monitoring modes with positive electrospray ionization. Results: The established method had a good linear relationship in the range of 0.0600-60.0μg/ml for caffeine, 0.0250-7.50μg/ml for theobromine and0.0150-4.50μg/ml for paraxanthine and theophylline. Conclusion: A HILIC-MS/MS method was developed and validated to determine caffeine and its major metabolites in plasma of premature infants.

Effects of serum branched-chain amino acids on nonalcoholic fatty liver disease and subsequent cardiovascular disease.

To reveal the role of branched-chain amino acids (BCAAs) in the development and progression of nonalcoholic fatty liver disease (NAFLD) and the effect on the incidence of subsequent cardiovascular disease. A total of 1302 subjects in the cohort study of the Huai'an Diabetes Prevention Program were divided into two groups according to whether NAFLD was present at baseline. The group without NAFLD at baseline was only followed up, and the group with NAFLD at baseline received diet and exercise interventions. Anthropometric and biochemical examinations were performed at baseline and at the end of 4years for all subjects. Serum BCAA (leucine, isoleucine, and valine) levels were measured by hydrophilic interaction chromatography-tandem mass spectrometry. The associations of baseline serum BCAA levels with the risk for NAFLD, coronary heart disease (CHD), and cardiovascular events (CVEs) after 4years were further evaluated. (1) At baseline and after the 4-year follow-up, baseline serum leucine, valine, and total BCAAs in the NAFLD group were significantly higher than those in the non-NAFLD group (p < 0.05). (2) According to whether NAFLD was present at baseline and after follow-up, all subjects were divided into four groups, including the control group, new case group, improvement group, and unchanged group. There was no significant difference in baseline BCAAs levels between the new case group and the improvement group (p > 0.05). (3) Risk factors for the occurrence and development of NAFLD were analysed by a multiple logistic regression model according to whether NAFLD existed at baseline. Serum leucine (OR = 1.058, 95% CI 1.005-1.114, p = 0.033) and total BCAAs (OR = 1.023, 95% CI 1.001-1.046, p = 0.045) were independent risk factors for new-onset NAFLD. Serum valine (OR = 1.131, 95% CI 1.043-1.226, p = 0.003), and total BCAAs (OR = 1.040, 95% CI 1.003-1.078, p = 0.035) were independent risk factors showing that NAFLD could not be reversed. (4) The cross-table Chi-square test showed that the incidence of both CHD and CVEs was significantly highest in the new case group (p < 0.05). (5) After adjusting for confounding factors, baseline isoleucine, valine, and BCAA levels were independently associated with new-onset CHD in subjects with or without NAFLD at baseline (p < 0.05). High BCAA levels exacerbate the risk of CHD and CVEs by influencing the occurrence and progression of NAFLD. However, lifestyle interventions could reverse the risk of NAFLD, CHD and CVEs associated with BCAAs.

Simultaneous Determination of Metabolites Related to Arginine Metabolism in Rat plasma by Hydrophilic Interaction Chromatography-Tandem Mass Spectrometry.

Arginine and its metabolites play important roles in pain and analgesia. This study aimed to develop a comprehensive quantification method for amino acids and metabolites related to arginine metabolism in rat plasma by hydrophilic interaction chromatography-tandem mass spectrometry (HILIC-MS/MS). Rat plasma was diluted to reduce the matrix effect and deproteinized with acetonitrile. The analytes were separated on a Syncronis HILIC column with a gradient elution. MS analysis was performed in positive ion mode with an electrospray ionization source using multiple reaction monitoring technology. All calibration curves for the 10 analytes showed good linear regression (R2 > 0.99). The limits of detection (LODs) were in the range of 0.9-13.4μg/L. The established method was validated for intra-day and inter-day precisions (relative standard deviation [RSDs] < 6.21%) and accuracy (average recovery ranged from 87.34% to 100.35% with the RSD values less than 11.41%). This method was successfully applied to characterize dynamic alterations in the plasma of rats with neuropathic pain and thus provide service to explore the mechanism of action between metabolite changes and clinical disease.

Metabolomic navigated Citrus waste repurposing to restore amino acids disorder in neural lesion

The fruit juice food industry produces huge waste annually, mainly Citrus peel and seeds. We investigated their chemical composition using hydrophilic interaction chromatography (HILIC-) and reverse phase liquid chromatography tandem mass spectrometry (RPLC-MS/MS), revealing 277 compounds, mainly containing flavonoids and limonoids. As the primary representative component in Citrus waste, limonin was selected to be explored new bio-functions. We applied Zebrafish larvae to study the metabolomic response invoked by limonin. The differential metabolites (DMs) varied depending on the exposing concentration of limonin. Enrichment analysis indicated that the identified DMs related to inflammation and neurologic disorders, including epilepsy which were newly discovered for limonoids and Citrus waste. Limonin was found to restore amino acids disorder to take neuroprotection against epilepsy. Our findings provided a new bio-function and purpose for Citrus waste and limonoids. Also, we demonstrated a concise case to repurpose food waste for new applications under metabolome investigation.

Attenuation of doxorubicin-induced oxidative damage in rat brain by regulating amino acid homeostasis with Astragali Radix.

The nervous system disorders caused by doxorubicin (DOX) are among the severe adverse effects that dramatically reduce the quality of life of cancer survivors. Astragali Radix (AR), a popular herbal drug and dietary supplement, is believed to help treat brain diseases by reducing oxidative stress and maintaining metabolic homeostasis. Here we show the protective effects of AR against DOX-induced oxidative damage in rat brain via regulating amino acid homeostasis. By constructing a clinically relevant low-dose DOX-induced toxicity rat model, we first performed an untargeted metabolomics analysis to discover specific metabolic features in the brain after DOX treatment and AR co-treatment. It was found that the amino acid (AA) metabolism pathways altered most significantly. To accurately characterize the brain AA profile, we established a sensitive, fast, and reproducible hydrophilic interaction chromatography-tandem mass spectrometry method for the simultaneous quantification of 22 AAs. The targeted analysis further confirmed the changes of AAs between different groups of rat brain. Specifically, the levels of six AAs, including glutamate, glycine, serine, alanine, citrulline, and ornithine, correlated (Pearson |r| > 0.47, p < 0.05) with the brain oxidative damage that was caused by DOX and rescued by AR. These findings present that AAs are among the regulatory targets of DOX-induced brain toxicity, and AR is a promising therapeutic agent for it.

Performance Characteristics of Refined LC-FLD and HILIC-MS/MS Methods for the Determination of Paralytic Shellfish Toxins in Shrimp, Whelk, and Crab.

Paralytic Shellfish Poison (PSP) toxins have been reported in non-bivalve shellfish species, including crustaceans and gastropods. Routine surveillance of these species is currently conducted in parts of England. To date, detection methods have not been validated for these matrices. Validation is required to ensure the test is fit for purpose, to give greater confidence in any results generated and ultimately facilitates accreditation. The aim was to test and validate two independent PSP toxin detection methods previously validated for bivalve shellfish matrices, for applicability to commercial non-bivalve species of interest. Matrices were shrimp (Crangon crangon), common whelk (Buccinum undatum), and edible crab (Cancer pagurus). The two methods assessed were the pre-column oxidation high-performance liquid chromatography-fluorescence detection AOAC 2005.06 Official Method of analysis and an internationally validated hydrophilic interaction chromatography-tandem mass spectrometry method. Brown and white crab meat were assessed separately. A refined extraction protocol was implemented with an increased solvent to sample ratio. The same extraction protocol was utilized for both methods, allowing both methods to be run simultaneously. Method sensitivity, recovery, repeatability, and method uncertainty were characterized in all matrix/toxin combinations. Overall, both methods performed similarly to that previously reported in bivalve mollusks. Acceptability of the majority of toxin/matrix combinations was evidenced through comparison of method performance characteristics against specific performance criteria, including Horwitz ratio values. Both PSP toxin detection methods were found to provide acceptable performance for the monitoring of shrimp, whelk, and crab species. Two PSP toxin detection methods have been single-laboratory validated successfully for three non-bivalve shellfish species.

Alteration of the goat milk glycoproteins N/O-glycome at different lactation stages

Goat milk oligosaccharides represent an unexplored multi-functional ingredient for the dairy industry. Here, we qualitatively and quantitatively compared the N/O-glycome at different lactation stages via online hydrophilic interaction chromatography-tandem mass spectrometry. Complex N-glycans and high mannose N-glycans constituted 82.1% and 17.9% of the glycan pool, respectively. N-glycans with isomers containing non-bisected antenna complex structures accounted for 30.8%. N-glycans modified with Neu5Ac, Neu5Gc and fucosylated were 3.7%, 5.3% and 35.3%. The triantennary trifucosylated complex N-glycan (H5N5F3) was reported for the first time. A comparison between colostrum and mature milk revealed a 1.20-fold decrease in total N-glycans and 1.66-fold decrease in fucosylation with ongoing lactation, echoing the trend in human milk. Similarly, Neu5Ac- and Neu5Gc-modified sialylation decreased by 1.69 and 3.62 times, respectively. In the O-glycome, 46.2% of structures were O-linked core 1, 23.1% were O-linked core 2, 7.7% were O-linked core 3 and core 4. As lactation progressed, overall O-glycans content decreased by 1.26-fold. Unlike human milk, Neu5Ac- and Neu5Gc-modified sialylation increased by 4.4 and 2 times, respectively. These findings will facilitate research on the structure-function relationship of goat milk oligosaccharides and the development of formula food targeting different age groups.

Quantitative and comparative study of plasmalogen molecular species in six edible shellfishes by hydrophilic interaction chromatography mass spectrometry

Shellfishes contain plasmalogens correlating to the functions of brain, heart, etc. Herein, a mild acid hydrolysis and hydrophilic interaction chromatography (HILIC) tandem mass spectrometry method was developed for analyzing plasmalogens in six shellfish species. A total of 19 plasmalogen molecular species were successfully identified, including nine phosphatidylcholine plasmalogen (plasPC), seven phosphatidylethanolamine plasmalogen (plasPE), and three phosphatidylserine plasmalogen (plasPS). The quantitative results indicated that mussel (32 μg·mg−1) possessed the highest content of plasmalogens, followed by oyster (21 μg·mg−1) and razor clam (15 μg·mg−1). The statistic models showed that the plasPE P-18:0/20:5 (m/z 748), plasPE P-16:0/22:2 & P-18:0/20:2 (m/z 754) and plasPS were the most contributing difference between shellfishes. The results indicated that this method was sensitive and precise to determine plasmalogens in shellfish, and mussel was demonstrated to be a good choice for the large-scale preparation of plasmalogens.

Quantitative determination of colistin A/B and colistin methanesulfonate in biological samples using hydrophilic interaction chromatography tandem mass spectrometry.

Colistin (polymyxin E) is a polycation antibiotic which is increasingly used (administered as colistin methanesulfonate, CMS) as a salvage therapy in critically ill patients with multidrug resistant Gram-negative infections. Even though colistin has been used for more than 50 years, its metabolic fate is poorly understood. One of the current challenges for studying the pharmacokinetics (PK) is the precise and accurate determination of colistin in in vitro and in vivo studies. In the present study, we developed and validated a series of sensitive and robust liquid chromatography tandem mass spectrometry (LC-MS/MS) methods for analysing biological samples obtained from in vitro and in vivo disposition assays. After a zinc acetate-mediated precipitation, hydrophilic-lipophilic-balanced solid phase extraction (HLB-SPE) was used for the extraction of colistin. The compounds were retained on a hydrophilic interaction liquid chromatography (HILIC) column and were detected by MS/MS. CMS was quantified by determining the produced amount of colistin during acidic hydrolysis. The developed methods are sensitive with lower limits of quantification varying between 0.009 μg/mL and 0.071 μg/mL for colistin A, and 0.002 μg/mL to 0.013 μg/mL for colistin B. The intra- and inter-day precision and accuracy were within ±15%. Calibration curves of colistin were linear (0.063 μg/mL to 8.00 μg/mL) within clinically relevant concentration ranges. Zinc acetate-mediated precipitation and the use of a HILIC column were found to be essential. The developed methods are sensitive, accurate, precise, highly efficient and allow monitoring colistin and CMS in biological samples without the need for an internal standard.

Aminopropyl hybrid silica monolithic capillary containing mesoporous SBA-15 particles for in-tube SPME-HILIC-MS/MS to determine levodopa, carbidopa, benserazide, dopamine, and 3-O-methyldopa in plasma samples

This manuscript describes an automated method that combines the selectivity of in-tube solid-phase microextraction (in-tube SPME) with the sensitivity of liquid hydrophilic interaction chromatography-tandem mass spectrometry (HILIC-MS/MS) to determine levodopa, carbidopa, benserazide, dopamine, and 3-O-methyldopa in plasma samples. An aminopropyl hybrid silica monolithic capillary containing mesoporous Santa Barbara Amorphous (SBA-15) particles for in-tube SPME was prepared by sol-gel reactions (one step). The organic–inorganic hybrid silica monolithic capillary was reused over fifty times without significant extraction efficiency loss, which attested to its robustness. The FTIR spectra displayed readily identifiable peaks that were characteristic of aminopropyl groups. SEM and TEM micrographs showed that the monolithic phase consisted of a homogeneous, continuous, and mesoporous skeleton composed of interconnected ordered particles. The developed method afforded adequate linearity at concentrations ranging from the lower limit of quantification (1.2–170 ng mL−1) to the upper limit of quantification (2000 ng mL−1), and the coefficients of correlation (r) were higher than 0.99. The method was validated with intra-day and inter-day precision and accuracy. In-tube SPME-HILIC-MS/MS was successfully developed to determine levodopa, carbidopa, benserazide, dopamine, and 3-O-methyldopa in plasma samples obtained from patients with Parkinson’s Disease for therapeutic drug monitoring purposes.

Determination of Moniliformin in Vegetable Oil by Solid-Phase Extraction–Hydrophilic Interaction Chromatography–Tandem Mass Spectrometry

An analytical method based on solid-phase extraction–hydrophilic interaction chromatography–tandem mass spectrometry (SPE–HILIC–MS/MS) has been developed to quantify moniliformin in vegetable oil. The sample was extracted by methanol and purified by an Oasis MAX solid-phase extraction column. Following purification, it was separated by a Waters HILIC hydrophilic interaction column. Acetonitrile and 0.5 mM ammonium acetate solution were used as a mobile phase for gradient elution. An electrospray-negative ion multiple-reaction monitoring mode was used to quantify moniliformin by external standard method. In the mass concentration range of 0.1–10 μg/L, the correlation coefficient of moniliformin was 0.9997. The detection limit of this method was 0.03 μg/L, and the limit of quantification was 0.1 μg/L. When the three levels of 1.0, 2.0 and 5.0 μg/kg moniliformin were added, the recovery was 84.6%–98.2% with a relative standard deviation of 1.3%–5.1%. The results indicated that this method is simple, fast, sensitive, reproducible, and can be used to determine moniliformin in vegetable oil.

A simple, versatile, and automated pulse-diffusion-focusing strategy for sensitive milliliter-level-injection HILIC-MS/MS analysis of hydrophilic toxins

The measurement of trace hydrophilic toxins in complex aqueous-rich matrices is a daunting challenge. To address this analytical bottleneck, pulse diffusion focusing (PDF), a novel sample injection technique for hydrophilic interaction chromatography-tandem mass spectrometry (HILIC-MS/MS), was developed. Theoretical and experimental investigations of the mechanism and key parameters revealed that the pulse-injection approach, assisted by solvent diffusion, efficiently solved the volume overload problem. This milliliter-level-injection HILIC-MS/MS technique was reported for the first time herein, and provided a significant enhancement in sensitivity compared to the conventional injection method, in addition to being an efficient approach to address the solvent incompatibility of off-line sample preparation and HILIC. The automated PDF-HILIC-MS/MS system was obtained by slightly modifying a commercial LC–MS/MS instrument in an easy and economical manner. The efficiency of the system was demonstrated through the detection of trace tetrodotoxin contents in plasma and urine samples. Low limits of detection (i.e., 0.65 and 2.2 ng·mL−1) were obtained using the simplified sample preparation method. The recoveries were in the range 91–113.3 % with intra-day and inter-day precisions of ≤9.6 %. Further experimental results proved the method to be versatile for various hydrophilic toxins.

Application of Stable Isotope Tracing to Elucidate Metabolic Dynamics During Yarrowia lipolytica α-Ionone Fermentation.

SummaryTargeted metabolite analysis in combination with 13C-tracing is a convenient strategy to determine pathway activity in biological systems; however, metabolite analysis is limited by challenges in separating and detecting pathway intermediates with current chromatographic methods. Here, a hydrophilic interaction chromatography tandem mass spectrometry approach was developed for improved metabolite separation, isotopologue analysis, and quantification. The physiological responses of a Yarrowia lipolytica strain engineered to produce ∼400 mg/L α-ionone and temporal changes in metabolism were quantified (e.g., mevalonate secretion, then uptake) indicating bottleneck shifts in the engineered pathway over the course of fermentation. Dynamic labeling results indicated limited tricarboxylic acid cycle label incorporation and, combined with a measurable ATP shortage during the high ionone production phase, suggested that electron transport and oxidative phosphorylation may limit energy supply and strain performance. The results provide insights into terpenoid pathway metabolic dynamics of non-model yeasts and offer guidelines for sensor development and modular engineering.

Accurate, sensitive and rapid determination of perchlorate in tea by hydrophilic interaction chromatography-tandem mass spectrometry.

Perchlorate is an environmental contaminant interrupting thyroid hormone production, and perchlorate in tea has raised wide concern recently. In this study, an accurate method was developed for the determination of perchlorate in tea using hydrophilic interaction chromatography-tandem mass spectrometry and a simplified QuEChERS procedure. The method utilized a zwitterion HILIC column for separation, and the optimal gradient eluents consisted of acetonitrile and aqueous solution with 0.1% formic acid and 20 mmol L-1 ammonium formate. Calibration curves were fitted by the quadratic model with 1/x weight instead of the linear model. As perchlorate was only partially extractable when using acetonitrile or methanol as the extraction solvent, acetonitrile/water (1 : 1, v/v) was chosen to extract perchlorate from tea samples. Graphitized carbon black was used as the dispersive solid phase extraction sorbent to clean up tea extracts. The method exhibited satisfactory accuracy with recoveries of 81.4-100.9% and relative standard deviations of 1.3-14.5% for green and black teas. The limit of quantitation was 0.005 mg kg-1, while the limits of detection were 0.0011 mg kg-1 for green tea and 0.0013 mg kg-1 for black tea, indicating an excellent sensitivity of this method. A 100% positive rate of perchlorate was found in 100 real tea samples, and the concentrations ranged from 0.0030 mg kg-1 to 0.78 mg kg-1. This accurate, sensitive and rapid method would be suitable for monitoring, risk assessment and source identification of perchlorate in tea.

2296-PUB: Serum Metabolome Reflects the Status of Glucose Metabolism of Japanese Pregnant Women

Objective: We reported that some metabolites during early pregnancy could predict subsequent development of gestational diabetes. In this study, to elucidate the metabolic status of pregnant women associated with glucose metabolism, we examined the association of the metabolomic profile and the glycohemoglobin (A1c) levels in Japanese pregnant women. Research Design and Methods: This study was conducted with participants enrolled by the Center of Chiba Unit of the Japan Environment and Children’s Study. Of those, 237 participants were enrolled in this study. Metabolite levels of their serum obtained during early pregnancy were analyzed using hydrophilic interaction chromatography tandem mass spectrometry. To examine the association between A1c levels and metabolic profiles, we performed least absolute shrinkage and selection operator (lasso) regression. The study protocol was approved by the Biomedical Research Ethics Committee of the Graduate School of Medicine, Chiba University. Results: Characteristics of participants were followed: age 32.0 (5.0) years, pre-gestational BMI 22.7 (4.6) kg/m2, A1c 5.09 (0.37) %. The association between their A1c levels and the metabolomic profiles, analyzed using lasso regression showed that nineteen metabolites were associated with A1c levels. Enrichment analysis revealed that the metabolites associated with A1c were related to ABC transporters, aminoacyl-tRNA biosynthesis, arginine and proline metabolism, glycine, serine and threonine metabolism, and metabolic pathway. Conclusion: Our results suggest the association of the serum metabolomic profile with the level of glycemia during pregnancy. Disclosure K. Sakurai: None. A. Eguchi: None. M. Watanabe: None. M. Yamamoto: None. C. Mori: None. Funding Japan Society for the Promotion of Science; Japan Ministry of the Environment

Simultaneous determination of cartap and its metabolite in tea using hydrophilic interaction chromatography tandem mass spectrometry and the combination of dispersive solid phase extraction and solid phase extraction

Risk assessment of cartap residue in tea should include the exposure of cartap and its metabolite due to rapid degradation of cartap into nereistoxin. Herein, a reliable method for determination of cartap and nereistoxin in tea was developed by hydrophilic interaction chromatography tandem mass spectrometry. Target compounds were extracted with water containing 1% formic acid and 5 mM ammonium formate. The use of dichloromethane effectively removed caffeine. Tea extracts were cleaned up by dispersive adsorbents of octadecylsilane and strong anion exchanger, then further purified using hydrophilic lipophilic balanced solid phase extraction cartridge. Isotopic internal standard was employed to calibrate the loss of analytes during sample preparation and compensate matrix effects. Method validation illustrated excellent linearity, with correlation coefficients (R2) higher than 0.999. Satisfactory recoveries of target compounds spiked in green tea, black tea and oolong tea ranged from 87.6% to 119.9% with intra- and inter-day precisions below 20%. Limits of quantification of cartap and nereistoxin were 10.0 μg kg−1, and limits of detection were 2.0 μg kg−1 for cartap and 4.0 μg kg−1 for nereistoxin. The developed method was applied to determine cartap and nereistoxin in thirty tea samples.

Simultaneous Quantification of 22 Glucosinolates in 12 Brassicaceae Vegetables by Hydrophilic Interaction Chromatography-Tandem Mass Spectrometry.

Glucosinolates, which are unique to Brassicaceae vegetables, have diverse biological activities, including antimicrobial, antioxidant, and anticancer actions. In this study, we applied hydrophilic interaction chromatography–tandem mass spectrometry (HILIC–MS/MS) to the simultaneous quantification of 22 glucosinolates in 12 Brassicaceae vegetables, including pak choi, choy sum, Chinese cabbage, cauliflower, cabbage, broccoli, Kai Lan, Brussels sprouts, rocket salad, daikon radish, red cherry radish, and watercress. Significant differences in concentration and composition of glucosinolates were observed among these vegetables. Cabbage had the highest level of total glucosinolates (μg/g dry weight: 19 551.2 ± 1317.7), whereas Kai Lan had the lowest level (7611.3 ± 868.4). Aliphatic and indole glucosinolates were the major components in the 12 vegetables ranging from 76 to 100%, except watercress (37%). On the basis of the content of glucosinolates, the 12 vegetables were well distinguishable and classified according to their morphological taxonomy. This study presents a HILIC–MS/MS approach for quantification of glucosinolates, and demonstrates the potential of glucosinolate profiles for Brassicaceae species identification.

Exploration of predictive metabolic factors for gestational diabetes mellitus in Japanese women using metabolomic analysis.

Aims/IntroductionWe aimed to explore novel predictive markers for gestational diabetes mellitus using metabolomic analysis in pregnant Japanese women.Materials and MethodsWe carried out a case–control study with a cohort of participants enrolled during the first or early second trimester in the Center of Chiba Unit of the Japan Environment and Children's Study. Participants were classified as either gestational diabetes mellitus cases or matched controls based on age, body mass index and parity. Metabolite levels of their serum and urine obtained randomly before the diagnosis of gestational diabetes mellitus were analyzed using hydrophilic interaction chromatography tandem mass spectrometry. Orthogonal projections to latent structures discriminant analysis was carried out to investigate metabolome profiles for the different groups. Metabolites with a variable importance in projection value of >1.5 were identified as potential markers.ResultsIn total, 242 participants were enrolled in the study, of which 121 were cases. The R2X, R2Y and Q2 parameters for the discrimination ability of the resulting models were 0.388, 0.492 and 0.45 for serum, and 0.454, 0.674 and 0.483 for urine, respectively. We finally identified three metabolites in serum and 20 in urine as potential biomarkers. Glutamine in serum and ethanolamine and 1,3‐diphosphoglycerate in urine showed >0.8 area under the receiver operating characteristic curves.ConclusionsThe present study identified serum and urine metabolites that are possible predictive markers of subsequent gestational diabetes mellitus in Japanese women. Further studies are required to elucidate their efficacy.