High Performance Liquid Chromatography-high Resolution Research Articles

Evaluation of the toxicological effects of Neltuma alpataco (Prosopis alpataco) pod alkaloid extract

The pods of Neltuma spp. have shown potential as a source of protein and energy in livestock. However, prolonged consumption of some of these species can lead to neurological symptoms in ruminants. This study aimed to determine the alkaloid content, as well as the in vitro and in vivo effects of an alkaloid-enriched extract (AEE) from N. alpataco pods. High performance liquid chromatography-high resolution mass spectrometry (HPLC-HRMS) identified juliprosine and juliprosopine as primary alkaloids, with juliprosine being most abundant. AEE from N. alpataco demonstrated dose-dependent cytotoxicity on glioma cells after 48 h, with a 50% cytotoxic concentration (CC50) of 24.69 μg/mL. However, the release of LDH was observed only at the highest tested concentration, indicating cellular damage. Further examination through phase-contrast microscopy and dual acridine orange/ethidium bromide fluorescence staining revealed morphological changes consistent with an apoptotic mechanism of cell death, ultimately leading to secondary necrosis. Finally, the LD50 after intraperitoneal injection in mice was determined to be 12.98 mg/kg. Taken together, these findings demonstrated for the first time the in vivo and in vitro toxicity of the AEE from N. alpataco pods.

Study on the production of Sophorolipid by Starmerella bombicola yeast using fried waste oil fermentation.

Sophorolipids (SLs) are surface active compounds that have excellent surface-lowering properties. SLs were produced by Starmerella bombicola (CGMCC1576) yeast with sunflower seed oil, fried waste oil, cooked tung oil and raw tung oil used as hydrophobic carbon sources. The results showed that the strain could use sunflower seed oil and fried waste oil as hydrophobic carbon sources to produce SLs, and the yields were 44.52 and 39.09 gl-1. It could not be used as cooked tung oil and raw tung oil. The analysis by high-performance liquid chromatography/high resolution mass spectrometry (HPLC-MS/MS) showed that the main composition and structure of SLs produced by fermentation using fried waste oil were similar to that of sunflower seed oil as hydrophobic carbon source. The yield of SLs was the highest when the fried waste oil was used as hydrophobic carbon source, glucose (8%), waste oil (6%) and yeast (0.3%). When fried waste oil was used as a hydrophobic carbon source in a parallel 4-strand fermentation tank (FT), the combination with the largest yield and the most cost saving was that 3% of fried waste oil was added into the initial medium, and another 3% was again added after 72 h of fermentation. The total yield of SLs was 121.28 gl-1, and the yield of lactone SLs was 48.07 gl-1.

A-031 Global Discovery of Serological Metabolome Uncovers Unique Molecular Signature for Early Onset of Type 2 Diabetes Mellitus: A Retrospective Study in Chinese Population

Abstract Background Type 2 diabetes mellitus (T2DM) is a multifaceted disorder affecting epidermic proportion at global scope. Defective insulin secretion by pancreatic β-cells and the inability of insulin-sensitive tissues to respond appropriately to insulin are the primary causes of T2DM. Early detection of people at high risk of developing T2DM is of great importance as interventions such as dietary control and intentional weight loss can effectively improve the insulin sensitivity and prevent the disease incidence. However, circulating biomarkers that are indicative of early diabetic onset at asymptomatic stage have not been well described. Here, we aim to identify new serological markers indicating early asymptomatic onset of T2DM by MS-based metabolomic discovery. Methods Three independent cohorts were assembled for the study. The discovery cohort included sera from 25 patients with T2DM and 25 healthy controls. The validation cohort included sera from 15 patients with T2DM and 15 healthy controls. The testing cohort included sera from 111 patients with pre-diabetes and 89 healthy controls. Untargeted approach was used to characterize the serological metabolome by high performance liquid chromatography-high resolution mass spectrometry. Results The univariate analysis identified 11 metabolites that were significantly altered in patients with T2DM across discovery and validation cohorts. After evaluated by the testing cohort, 8 of 11 metabolites, such as hexose, N-acetyl-glucosamine, and alpha-ketobutyrate, remained statistically significant in patients with prediabetes as compared to healthy controls. The multivariate analysis based on 8 significant metabolites revealed distinct patterns of metabolic perturbation at the early onset of T2DM. To gain mechanistic insights into the disease pathophysiology, pathway enrichment and correlation network analyses were conducted to unveil the molecular crosstalk underlying the disturbed metabolic network in the pathogenesis of T2DM. Conclusion Our study not only identified serological biomarkers detecting early asymptomatic onset of T2DM but also provided additional insights into the disease pathophysiology.

Multi-omics approach to understand the impact of sun exposure on an in vitro skin ecosystem and evaluate a new broad-spectrum sunscreen.

A reconstructed human epidermal model (RHE) colonized with human microbiota and sebum was developed to reproduce the complexity of the skin ecosystem in vitro. The RHE model was exposed to simulated solar radiation (SSR) with or without SPF50+ sunscreen (with UVB, UVA, long-UVA, and visible light protection). Structural identification of discriminant metabolites was acquired by nuclear magnetic resonance and metabolomic fingerprints were identified using reverse phase-ultra high-performance liquid chromatography-high resolution mass spectrometry, followed by pathway enrichment analysis. Over 50 metabolites were significantly altered by SSR (p < 0.05, log2 values), showing high skin oxidative stress (glutathione and purine pathways, urea cycle) and altered skin microbiota (branched-chain amino acid cycle and tryptophan pathway). 16S and internal transcribed spacer rRNA sequencing showed the relative abundance of various bacteria and fungi altered by SSR. This study identified highly accurate metabolomic fingerprints and metagenomic modifications of sun-exposed skin to help elucidate the interactions between the skin and its microbiota. Application of SPF50+ sunscreen protected the skin ecosystem model from the deleterious effects of SSR and preserved the physiological interactions within the skin ecosystem. These innovative technologies could thus be used to evaluate the effectiveness of sunscreen.

Inherited disorders of hemoglobin: A review of old and new diagnostic methods

The genetic regulation of hemoglobin is complex and there are a number of genetic abnormalities that result in clinically important hemoglobin disorders. Here, we review the molecular pathophysiology of hemoglobin disorders and review both old and new methods of diagnosing these disorders. Timely diagnosis of hemoglobinopathies in infants is essential to coordinate optimal life-saving interventions, and accurate identification of carriers of deleterious mutations allows for genetic counseling and informed family planning. The initial laboratory workup of inherited disorders of hemoglobin should include a complete blood count (CBC) and peripheral blood smear, followed by carefully selected tests based on clinical suspicion and available methodology. We discuss the utility and limitations of the various methodologies to fractionate hemoglobin, including cellulose acetate and citrate agar hemoglobin electrophoresis, isoelectric focusing, high-resolution high-performance liquid chromatography, and capillary zone electrophoresis. Recognizing that most of the global burden of hemoglobin disorders exists in low- and middle-income countries, we review the increasingly available array of point-of-care-tests (POCT), which have an increasingly important role in expanding early diagnosis programs to address the global burden of sickle cell disease, including Sickle SCAN, HemoTypeSC, Gazelle Hb Variant, and Smart LifeLC. A comprehensive understanding of the molecular pathophysiology of hemoglobin and the globin genes, as well as a clear understanding of the utility and limitations of currently available diagnostic tests, is essential in reducing global disease burden.

Stabilization of N-terminal cysteines in HPLC-HRMS quality control of peptide pools

Abstract Synthetic peptide pools are used in experimental and clinical immunology, for example, cancer immunotherapy. The analytical characterization is challenging due to the similarity of the peptides, usually only a pre-characterization of the single peptides is performed. However, regular quality control of the peptide mix would be highly desirable. Therefore, a high performance liquid chromatography-high resolution mass spectrometry (HPLC-HRMS) method for quality control of a peptide pool was developed. Peptides were synthesized and purified to &amp;gt; 90% each. The lyophilized single peptides were combined to a peptide pool containing 32 peptides and analyzed by HPLC-HRMS. Quantification is carried out UV detection, and identification is obtained by high resolution mass spectrometry. Before separation on a capillary reversed-phase column, alkylation of thiols was performed. Best results were obtained with a linear gradient from 96% of solvent A (water with 0.05% TFA), 4% of solvent B (acetonitrile with 0.04% TFA) to 56% of solvent A, 44% of solvent B in 100 minutes. Without sample preparation, peptides with N-terminal cysteine will yield almost no UV signal. This surprising effect could be eliminated using iodoacetic acid (IAC) as an alkylation reagent. The amount of IAC and reductant (Tris(2-carboxyethyl)phosphine (TCEP)) was carefully optimized to avoid overalkylation. By a combination of capillary HPLC and HRMS, a method could be developed for the quality control of complex synthetic peptide pools. In contrast to the analysis of single peptides, analyzing complex peptide pools requires the stabilization of cysteine-containing peptides. Good results were obtained using iodoacetic acid as an alkylation reagent.

Evaluation of the Antioxidant Activities and Phenolic Profile of Shennongjia Apis cerana Honey through a Comparison with Apis mellifera Honey in China

This study evaluates the phenolic profile as well as the antioxidant properties of Shennongjia Apis cerana honey through a comparison with Apis mellifera honey in China. The total phenolic content (TPC) ranges from 263 ± 2 to 681 ± 36 mg gallic acid/kg. The total flavonoids content (TFC) ranges from 35.9 ± 0.4 to 102.2 ± 0.8 mg epicatechin/kg. The correlations between TPC or TFC and the antioxidant results (FRAP, DPPH, and ABTS) were found to be statistically significant (p < 0.01). Furthermore, the phenolic compounds are quantified and qualified by high performance liquid chromatography-high resolution mass spectrometry (HPLC-HRMS), and a total of 83 phenolic compounds were tentatively identified in this study. A metabolomics analysis based on the 83 polyphenols was carried out and subjected to principal component analysis and orthogonal partial least squares-discriminant analysis. The results showed that it was possible to distinguish Apis cerana honey from Apis mellifera honey based on the phenolic profile.

Amitraz and Its Metabolites: Oxidative Stress-Mediated Cytotoxicity in HepG2 Cells and Study of Their Stability and Characterization in Honey.

The population decrease of bees that has been observed in recent years due to the Varroa destructor parasite may endanger the production of bee-products whose demand is on the rise. To minimize the negative effects caused by this parasite, the pesticide amitraz is commonly used by beekeepers. Based on these, the objectives of this work are to determine the toxic effects caused by amitraz and its metabolites in HepG2 cells, as well as its determination in honey samples and the study of its stability with different heat treatments commonly used in the honey industry and its relationship with the amount of 5-hydroxymethylfurfural (HMF) produced. Amitraz significantly decreased cell viability by MTT assay and total protein content (PC) assay, being more cytotoxic than its metabolites. Amitraz and its metabolites caused oxidative stress by Lipid Peroxidation (LPO) production and Reactive Oxygen Species (ROS) generation. Residues of amitraz and/or its metabolites were found in analyzed honey samples, with 2,4-Dimethylaniline (2,4-DMA) being the main metabolite confirmed by high-performance liquid chromatography-high resolution mass spectrometry (HPLC-QTOF HRMS). Amitraz and its metabolites resulted as unstable even at moderate heat treatments. Additionally, a positive correlation in terms of HMF concentration in samples and the severity of heat treatment was also observed. However, quantified amitraz and HMF were within the levels set in the regulation.

The tRNA-degradation pathway impacts the phenotype and metabolome of Arabidopsis thaliana: evidence from atipt2 and atipt9 knockout mutants

Isopentenyltransferases (IPTs), including adenosine phosphate-isopentenyltransferases (ATP/ADP-IPTs and AMP-IPTs) and tRNA‐isopentenyltransferases (tRNA-IPTs), are responsible for a rate-limiting step of cytokinin (CK) biosynthesis. tRNA-IPTs, which account for the synthesis of cis-Zeatin (cZ)-type CKs, are less understood and have often been thought to play a housekeeping role or have low activity during plant growth and development. Even with increasing evidence supporting the tRNA-degradation pathway and the role of cis-CK metabolites to perform important functions, a fulsome examination at the metabolome level where critical biological effects are normally absent of phenotypic expression, remains unpublished. Here, two Arabidopsis tRNA-IPT knockout mutants, atipt2 and atipt9, with independent disturbance of the pathway leading to cisCKs were investigated at the phenotype and metabolite levels at four stages of plant development: first leaf, inflorescence, siliques, and mature seed. Phenotypic deviations were noted in rosette diameter, number of non-rosette leaves, shoot height, flowering time, flower number, carotenoid content, trichome development, and above-ground fresh mass. Hormone profiling by high-performance liquid chromatography-high resolution tandem mass spectrometry (HPLC-HRMS/MS) showed that the atipt2 mutant accumulates lower total cisCKs in the first leaves and in siliques. The atipt9 mutant showed reduced total cisCKs in first leaves, but, during silique development, it had higher levels of cisCKs in than those of the wild type (WT) plants. Additionally, metabolite detection was performed via an untargeted approach using HPLC-HRMS. A total of 33 significant features differing in abundance between ipt mutants and the WT were putatively identified based on database search. Matched metabolites included those that participate in hormone cross-talk, fatty acid synthesis, seed set and germination, and in stress acclimation. Evidence indicates that cisCK production is important for plant growth and development, in ways distinct from CKs produced from de novo pathway.

Metabolic study of selective androgen receptor modulator LY2452473 in thoroughbred horses for doping control.

Since 2010, there has been an increasing number of adverse analytical findings related to selective androgen receptor modulators (SARMs) in competitive sports. It emphasizes the importance of comprehensive doping control analytical procedures that are capable of detecting SARM misuse. In this study, it is described how LY2452473, a SARM, was metabolized in thoroughbred horses after a single-dose oral administration and in vitro with equine liver microsome preparations. An investigation of the metabolism of LY2452473 in horses' urine, plasma, and hair matrices was carried out during the study. The plausible structures of the detected metabolites were postulated using high-performance liquid chromatography-high resolution mass spectrometry. Under the experimental conditions 15 metabolites (12 phase I and three conjugates of phase I) were detected (M1-M15). The major phase I metabolites identified were formed by hydroxylation. Side-chain dissociated and methylated metabolites were also detected. In phase II, the glucuronic acid and sulfonic acid conjugates of hydroxy LY2452473 were detected as the major metabolites. In vitro analysis has confirmed the presence of all metabolites found in vivo except for the methylated analogs M11 and M12. A peak concentration of LY2452473 (0.5pg/mg) in proximal hair segments was achieved 4 weeks after administration, according to hair analysis. Data obtained will aid in identifying LY2452473 and related substances faster. Furthermore, the results will assist in checking for the illegal use of these substances in competitive sports.

Lithium Difluorophosphate: A Boon for High Voltage Li Ion Batteries and a Bane for High Thermal Stability/Low Toxicity: Towards Synergistic Dual Additives to Circumvent this Dilemma.

The specific energy/energy density of state-of-the-art (SOTA) Li-ion batteries can be increased by raising the upper charge voltage. However, instability of SOTA cathodes (i. e., LiNiy Cox Mny O2 ; x+y+z=1; NCM) triggers electrode crosstalk through enhanced transition metal (TM) dissolution and contributes to severe capacity fade; in the worst case, to a sudden death ("roll-over failure"). Lithium difluorophosphate (LiDFP) as electrolyte additive is able to boost high voltage performance by scavenging dissolved TMs. However, LiDFP is chemically unstable and rapidly decomposes to toxic (oligo)organofluorophosphates (OFPs) at elevated temperatures; a process that can be precisely analyzed by means of high-performance liquid chromatography-high resolution mass spectroscopy. The toxicity of LiDFP can be proven by the well-known acetylcholinesterase inhibition test. Interestingly, although fluoroethylene carbonate (FEC) is inappropriate for high voltage applications as a single electrolyte additive due to rollover failure, it is able to suppress formation of toxic OFPs. Based on this, a synergistic LiDFP/FEC dual-additive approach is suggested in this work, showing characteristic benefits of both individual additives (good capacity retention at high voltage in the presence of LiDFP and decreased OFP formation/toxicity induced by FEC).

An Integrative Pharmacology-Based Strategy to Uncover the Mechanism of Zuogui Jiangtang Shuxin Formula in Diabetic Cardiomyopathy.

This study aimed to explore the mechanism of Zuogui Jiangtang Shuxin formula (ZGJTSXF) in the treatment of diabetic cardiomyopathy (DCM) by an integrative strategy combining serum pharmacochemistry, network pharmacology analysis, and experimental validation. An Ultra high performance liquid chromatography-high resolution mass spectrometry (UPLC-Q-Exactive-Orbitrap-MS) method was constructed to identify compounds in rat serum after oral administration of ZGJTSXF. A component-target network between the targets of ZGJTSXF ingredients and DCM was established using Cytoscape. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed to deduce ZGJTSXF-associated targets and pathways. The DCM model mice were treated with ZGJTSXF, and the predicted important signaling pathways were verified using quantitative PCR and Western blot. We identified 78 compounds in serum of medicated rats, which mainly included flavonoids, small peptides, nucleosides, organic acids, phenylpropanoids, alkaloids, phenanthrenequinones, iridoids, phenols, and saponins. Network pharmacology analysis revealed that ZGJTSXF may regulate targets including ALB, TNF, AKT1, GAPDH, VEGFA, EGFR, SRC, CASP3, MAPK3, JUN, and PI3K/AKT signaling pathway in the treatment of DCM. ZGJTSXF administration improved blood sugar levels, heart function, and cardiac morphological changes in DCM mice. Notably, ZGJTSXF inhibited cardiomyocytes apoptosis, which was associated with restored PI3K/Akt signaling and upregulated Bcl-2 and Bcl-xL proteins expression. Our preliminary results proposed the material basis and possible mechanisms of ZGJTSXF in treating DCM, which is related to the activation of the PI3K/AKT signaling pathway and apoptosis inhibition. These findings shed new light in developing ZGJTSXF-based therapeutics in treating DCM.

Features and Practical Aspects of Radiochemical Purity Determination of Receptor-Specific Lu-177 Radiopharmaceuticals as Exemplified by [&lt;sup&gt;177&lt;/sup&gt;Lu]Lu–PSMA-617

Radiochemical purity (RCP) is one of the key quality criteria for radiopharmaceuticals (RPs) used in clinical practice. The results of RCP measurements depend on the analytical technique, as well as detection parameters, which are specific to a particular analytical system. When reviewing literature data on the synthesis, pharmaceutical development, preclinical and clinical trials of the same radiopharmaceutical by different authors, one may note significant variability in the RCP values obtained. Hence, it is important to carefully select analysis parameters and study their influence on the results. The aim of the study was to compare previously published and self-developed procedures for RCP analysis of 177 Lu-RPs in terms of their efficiency in the detection and quantification of possible radiochemical impurities, as well as to determine the analytical system parameters that have a significant impact on the interpretation of the analysis results, using [ 177 Lu]Lu–PSMA-617 as a case study. Materials and methods : the study used samples of [ 177 Lu]Lu–PSMA-617 with a volume activity of lutetium-177 of 150–4800 MBq/mL, containing 10–100 µmol/L of the PSMA-617 precursor and 30 mmol/L of sodium acetate as a buffering solution (pH 4.5). The samples were tested by high-performance liquid chromatography (HPLC) and thin-layer chromatography in the conditions described in the literature and developed in the course of the work. Results : the study showed a significant influence of the chosen analytical procedure on the results of [ 177 Lu]Lu–PSMA-617 RCP assessment. The profile of possible radiochemical impurities requires the use of high-resolution HPLC techniques. Conclusions : the analytical procedure developed and applied by the authors is quite effective. The results of RCP assessment are influenced by the detection system parameters, such as the length and inner diameter of the flow cell and the means of analytical signal processing (peak extraction parameters, smoothing parameters, and noise subtraction/suppression). This fact necessitates validation considering the characteristics of a particular analytical system and demonstrates the need to assess interlaboratory precision in the framework of the implementation and verification of analytical procedures.

Quality control standardization of Indonesian noni fruit (Morinda citrifolia) extract and evaluation of their angiotensin-converting enzyme inhibitory activity

Morinda citrifolia fruit (Noni) has been used as a folk medicine in several countries. Noni possessed various pharmacological activities such as; anticancer, antidiabetic, antihypertensive, antarthritic, and antioxidants. The present study evaluated pharmacognostic properties, profiling of active constituent through High-Performance Thin Layer Chromatography (HPTLC) and Liquid Chromatography-High Resolution Mass Spectrometer (LC-HRMS) run for quantitative and qualitative phytochemical analysis and determining angiotensin-converting enzyme (ACE) inhibitor activity of Noni from three different locations. The physicochemical parameters of crude drugs and extracts met the requirement of Indonesian Herbal Pharmacopeia. Total phenol content was 2.16-3.08 mg GAE/g extract and total flavonoid content was 0.11 - 1.58 mg QE/g extract. HPTLC analysis revealed that scopoletin content in Noni was in the range of 0.44 - 0.51%. The results were also corresponding well with LC-HRMS fingerprint analysis. In addition, Noni fruit extract from Bogor potential exhibited activity in inhibiting ACE with an IC50 value of 206.26 µg/mL.

Identification of a novel bromoindazole synthetic cannabinoid analogue in seized e-cigarette liquid: ADB-BRINACA

A wide variety of new synthetic cannabinoids have emerged around the world in recent years, and because of this rapid emergence, the detection and monitoring of this class of abused drugs remain a challenge. In this study, a new cannabimimetic indazole-3-carboxamide derivative, N-(1-amino-3,3-dimethyl-1-oxobutan-2-yl)− 5-bromo-1 H-indazole-3-carboxamide, was identified from seized e-cigarette liquid samples and newly named as ADB-BRINACA by referring to the names of other known indazole-class synthetic cannabinoids. Structure identification was accomplished based on gas chromatography-mass spectrometry (GC-MS), high-performance liquid chromatography-high resolution quadrupole mass spectrometry (HPLC-QTOF-MS/MS), and nuclear magnetic resonance spectrometry (NMR). The concentration range of ADB-BRINACA in six e-cigarette liquid samples was found to be 2228–4203 mg/L using ERETIC 2, a quantitative NMR method, which is advantageous in the absence of a reference material. As there have been no chemical or pharmaceutical reports on ADB-BRINACA until now, this is the first report presenting a comprehensive analytical characterization of ADB-BRINACA.

Determination of Organic Nitrogen Compounds in Gasoline by Water-Assisted Dispersive Solid-Phase Extraction (DSPE) and High-Performance Liquid Chromatography-High-Resolution Mass Spectrometry (HPLC-HRMS)

For the identification of trace-level organic nitrogen compounds (NCs) in gasoline, a method consisting of water-assisted dispersive solid-phase extraction (DSPE) and high-performance liquid chromatography-high resolution mass spectrometry was developed in which mixed-mode cation exchange was used as the adsorbent. To increase the DSPE efficiency, water was dispersed into the gasoline to increase the pronation of NCs and enhance the ionic interaction with the adsorbent. After systematic optimization of DSPE conditions, including the adsorbent mass, sample volume, water to gasoline ratio, sample loading time, washing and elution solvents, sensitive determination of NCs (detection limits from 0.1 to 5 ng/L for the 14 analytes) was achieved by this method with a 100-fold enrichment. A number of unknown NC isomers were observed from the extracted ion chromatograms based on the accurate determination of mass-to-charge ratio (m/z), from which two sets of isomers at theoretical m/z of 122.09643 and 136.11208 were selected for identification. By careful elucidation of the product ion spectra of the selected ions, 16 NCs were putatively identified, in which six non-aniline homologs were reported for the first time.

Serological Phenotyping Analysis Uncovers a Unique Metabolomic Pattern Associated With Early Onset of Type 2 Diabetes Mellitus.

Background: Type 2 diabetes mellitus (T2DM) is a multifaceted disorder affecting epidemic proportion at global scope. Defective insulin secretion by pancreatic β-cells and the inability of insulin-sensitive tissues to respond effectively to insulin are the underlying biology of T2DM. However, circulating biomarkers indicative of early diabetic onset at the asymptomatic stage have not been well described. We hypothesized that global and targeted mass spectrometry (MS) based metabolomic discovery can identify novel serological metabolic biomarkers specifically associated with T2DM. We further hypothesized that these markers can have a unique pattern associated with latent or early asymptomatic stage, promising an effective liquid biopsy approach for population T2DM risk stratification and screening. Methods: Four independent cohorts were assembled for the study. The T2DM cohort included sera from 25 patients with T2DM and 25 healthy individuals for the biomarker discovery and sera from 15 patients with T2DM and 15 healthy controls for the testing. The Pre-T2DM cohort included sera from 76 with prediabetes and 62 healthy controls for the model training and sera from 35 patients with prediabetes and 27 healthy controls for the model testing. Both global and targeted (amino acid, acylcarnitine, and fatty acid) approaches were used to deep phenotype the serological metabolome by high performance liquid chromatography-high resolution mass spectrometry. Different machine learning approaches (Random Forest, XGBoost, and ElasticNet) were applied to model the unique T2DM/Pre-T2DM metabolic patterns and contrasted with their effectiness to differentiate T2DM/Pre-T2DM from controls. Results: The univariate analysis identified unique panel of metabolites (n = 22) significantly associated with T2DM. Global metabolomics and subsequent structure determination led to the identification of 8 T2DM biomarkers while targeted LCMS profiling discovered 14 T2DM biomarkers. Our panel can effectively differentiate T2DM (ROC AUC = 1.00) or Pre-T2DM (ROC AUC = 0.84) from the controls in the respective testing cohort. Conclusion: Our serological metabolite panel can be utilized to identifiy asymptomatic population at risk of T2DM, which may provide utility in identifying population at risk at an early stage of diabetic development to allow for clinical intervention. This early detection would guide ehanced levels of care and accelerate development of clinical strategies to prevent T2DM.

Serum untargeted lipidomics by UHPLC-ESI-HRMS aids the biomarker discovery of colorectal adenoma

BackgroundColorectal adenoma (CA) is an important precancerous lesion and early screening target of colorectal cancer (CRC). Lipids with numerous physiological functions are proved to be involved in the development of CRC. However, there is no lipidomic study with large-scale serum samples on diagnostic biomarkers for CA.MethodsThe serum lipidomics of CA patients (n = 50) and normal control (NR) (n = 50) was performed by ultra high performance liquid chromatography-high resolution mass spectrometry with electrospray ionization (UHPLC-ESI-HRMS). Univariate and multivariate statistical analyses were utilized to screen the differential lipids between groups, and combining the constituent ratio analysis and diagnostic efficiency evaluation by receiver operating characteristic (ROC) curve disclosed the potential mechanism and biomarkers for CA.ResultsThere were obvious differences in serum lipid profiles between CA and NR groups. Totally, 79 differential lipids were selected by criterion of P < 0.05 and fold change > 1.5 or < 0.67. Triacylglycerols (TAGs) and phosphatidylcholines (PCs) were the major differential lipids with ratio > 60%, indicating these two lipid metabolic pathways showed evident disequilibrium, which could contribute to CA formation. Of them, 12 differential lipids had good diagnostic ability as candidate biomarkers for CA (AUC ≥ 0.900) by ROC analysis.ConclusionsTo our knowledge, this is the first attempt to profile serum lipidomics and explore lipid biomarkers of CA to help early screening of CRC. 12 differential lipids are obtained to act as potential diagnostic markers of CA. PCs and fatty acids were the main dysregulated biomarkers for CA in serum.

Saliva Metabolomics in Dry Mouth Patients with Head and Neck Cancer or Sjögren's Syndrome.

The etiology of dry mouth conditions is multi-faceted. Patients radiated after head and neck cancer (HNC) and those with primary Sjögren’s syndrome (pSS) share many of the same symptoms despite different causes. With the aim of better understanding the pathophysiology and biochemical processes behind dry mouth with different etiologies, we investigated the metabolic profile of 10 HNC patients, 9 pSS patients and 10 healthy controls using high-performance liquid chromatography-high resolution mass spectrometry (HPLC-MS) metabolomics. Principal component analysis (PCA) revealed different metabolic profiles when comparing all subjects included in the study. Both patient groups showed higher ratios of several pyrimidine nucleotides and nucleosides when compared to controls. This finding may indicate that purinergic signaling plays a role in dry mouth conditions. Moreover, significantly increased levels of DL-3-aminoisobutyric acid were found in HNC patients when compared to controls, and a similar tendency was observed in the pSS patients. Furthermore, a dysregulation in amino acid metabolism was observed in both patient groups. In conclusion, metabolomics analysis showed separate metabolic profiles for HNC and pSS patients as compared to controls that could be useful in diagnostics and for elucidating the different pathophysiologies. The demonstrated dysregulation of pyrimidine nucleotides and levels of metabolites derived from amino acids in the patient groups should be studied further.

Acute effect of oat β-glucan on the bioavailability of orange juice flavanones

The impact of β-glucan on the bioavailability of orange juice (OJ) flavanones was investigated in a randomised controlled trial. Volunteers consumed 500 mL of OJ without or with either 3 g (OB-3) or 6 g (OB-6) of β-glucan. Urine samples, collected 12 h before and over a 0–24 h period post-supplementation, were analysed by high-performance liquid chromatography-high resolution mass spectrometry. The overall 0–24 h urinary excretion of the 17 flavanone metabolites identified and quantified in urine after OJ ingestion corresponded to 29.7 µmol, and 25.0 and 9.3 µmol, respectively, after OB-3 and OB-6 intake. This corresponds to 9.3, 7.9, and 2.9% recoveries of the 318 µmol of the ingested flavanones. The acute ingestion of OJ with 6 g, but not 3 g of β-glucan led to a significant reduction (p < 0.05) in the excretion of flavanone metabolites compared with consumption of OJ alone.