Ultra-performance Liquid Chromatography-tandem Mass Research Articles

Pharmacokinetic Interaction Between Olaparib and Regorafenib in an Animal Model

Abstract: Background: Olaparib (OLA) and regorafenib (REG) are metabolized by the CYP3A4 isoenzyme of cytochrome P450. Both drugs are also substrates and inhibitors of the membrane transporters P-glycoprotein and BCRP. Therefore, the potential concomitant use of OLA and REG may result in clinically relevant drug–drug interactions. Knowledge of the influence of membrane transporters and cytochrome P450 enzymes on the pharmacokinetics of drugs makes it possible to assess their impact on the efficacy and safety of therapy. Purpose: The study aimed to evaluate the bilateral pharmacokinetic interactions of OLA and REG and its active metabolites after a single administration in healthy rats. Methods: The study was performed in male Wistar rats (n = 24) randomly divided into three groups: one study group, IREG+OLA (n = 8), received REG with OLA, and two control groups, IIREG (n = 8) and IIIOLA (n = 8), received REG and OLA, respectively. The concentrations of OLA, REG, REG-N-oxide (M-2), and N-desmethyl-REG-N-oxide (M-5) were determined by ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS). The values of the pharmacokinetic parameters of OLA, REG, M-2, and M-5 were determined by non-compartmental analysis with linear interpolation. Results: After OLA administration, the pharmacokinetic parameters of REG (AUC0–∞, tmax, and t0.5) increased significantly by 3.38-, 2.66-, and 1.82-fold, respectively. On the other hand, REG elimination parameters, i.e., kel and Cl/F, were significantly reduced in the study group by 1.77- and 1.70-fold, respectively. In the study group, Cmax and AUC0-t values were also 7.22- and 8.86-fold higher for M-2 and 16.32- and 17.83-fold higher for M-5, respectively. The Metabolite M-2/Parent and Metabolite M-5/Parent ratios for Cmax and AUC0-t increased by 6.52-, 10.74-, 28-, and 13-fold, respectively. After administration of OLA with REG, the Cmax, AUC0-t, and AUC0-∞ of OLA increased by 2.0-, 3.4-, and 3.4-fold, respectively, compared to the control group. Meanwhile, Cl/F and Vd/F of OLA were significantly decreased in the presence of REG. Conclusions: OLA was shown to significantly affect the pharmacokinetics of REG and its active metabolites M-2 and M-5 in rats after co-administration of both drugs. There was also a significant effect of REG on the pharmacokinetics of OLA, which may have clinical relevance. The AUC ratios (study group/control group) were 3.41 and 3.39 for REG and OLA, respectively, indicating that REG and OLA were moderate inhibitors in this preclinical study. The results obtained need to be confirmed in clinical studies. This study may provide guidance on the safety of using both drugs in clinical practice.

A Metabolomic Analysis of Tomato Fruits in Response to Salt Stress

Salt stress affects all stages of tomato growth and development, reducing tomato yield, but moderate salt stress improves tomato quality. To gain a deeper understanding of the effect of salt stress on tomato fruits, a widely targeted metabolomic method based on ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) was used to analyze tomato fruits under three different soil salt contents (CK: 0.05 g·kg−1; MS: 3 g·kg−1; and HS: 6 g·kg−1). A total of 847 metabolites were detected in tomato fruit under salt stress, including six primary metabolites such as lipids, amino acids and their derivatives, sugars, nucleotides and their derivatives, and organic acids, and nine secondary metabolites such as phenolic acids, alkaloids, and flavonoids. There were 54 different metabolites in the three treatments, mainly flavonols and phenolamines. Dopamine, galactotol, and mannitol were not detected in the control group, but their contents were higher in the salt treatment. KEGG analysis showed that the differential metabolites were mainly concentrated in flavonoid biosynthesis, betaine metabolism, sulfur metabolism, and galactose metabolism. This study provides a theoretical basis for the regulation of tomato quality through salt stress.

The amino acid metabolism pathway of peripheral T lymphocytes and ketamine-induced schizophrenia-like phenotype.

The intricate interplay between peripheral adaptive immune cells and the central nervous system (CNS) has garnered increasing recognition. Given that alterations in cell quantities often translate into modifications in metabolite profiles and that these metabolic changes can potentially traverse the bloodstream and enter the CNS, thereby modulating the progression of mental illnesses, we sought to explore the metabolic profiles of peripheral immune cells in a ketamine-treated mouse model of schizophrenia. We used flow cytometry to scrutinize the alterations in peripheral adaptive immune cells in a ketamine-induced schizophrenia mouse model. Subsequently, we implemented an untargeted metabolomic approach with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to detect the metabolite profiles of peripheral abnormal lymphocytes and identify differential metabolites present in plasma. We then employed targeted metabolomics using UPLC-MS/MS to quantify the common differential metabolites detected in mouse plasma. Flow cytometry analysis detected a notable increase in the count of peripheral CD3+ T cells in a ketamine-induced schizophrenia mouse model. Subsequent untargeted metabolomics analysis revealed that the amino acid metabolism pathway underwent substantial alterations. A detailed quantification of 22 amino acid profiles in the peripheral plasma indicated significant elevation in the levels of glycine, alanine, asparagine, and aspartic acid. Our ongoing research has yet to conclusively identify the precise amino acid metabolism pathway that serves as the pivotal factor in the manifestation of the schizophrenia-like phenotype induced by ketamine. The peripheral amino acid metabolism pathway is involved in the ketamine-induced schizophrenia-like phenotype. The metabolic profile of peripheral immune cells could provide accurate biomarkers for the diagnosis and treatment of psychiatric diseases.

LC-MS/MS method for the simultaneous determination of six phenolic acids in rat plasma and hypoglycemic effects in normal and T2DM rats after oral administration of Eleutherococcus senticosus fruits extract

AbstractEleutherococcus senticosus (Rupr. & Maxim.) Maxim. fruits (ESF) is a natural health food that has been widely used in traditional medicine. This study aimed to establish and validate a rapid and sensitive method for determining of six phenolic acids in plasma of normal and T2DM rats by ultra-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS). Separation and quantification were performed on a Waters HSS T3 column (100 × 2.1 mm, 1.8 μm) with water (A) and methanol (B). The electrospray ionization (ESI) source was operated in negative ionization mode. Multiple reaction monitoring (MRM) was used to quantitatively analyzed hyperoside m/z 463.2→301.1, caffeic acid m/z 179.1→135.1, chlorogenic acid m/z 353.2→190.9, gallic acid m/z 169.0→125.1, protocatechuic acid m/z 153.1→109.1, and isofraxidin m/z 221.1→191.0, respectively. The hypoglycemic effect of ESF was verified by determining blood glucose and lipid levels in a type 2 diabetes model. This study laid the foundation for exploring ESF clinical guidelines in the future.

Removal residual, commercial processing factor and risk assessment of four fungicides in orange fruit and various processing by-products

The residual behaviors and processing factors of four fungicides (prochloraz, kasugamycin, oxine-copper and fenaminstrobin) in orange fruit and various processing by-products were evaluated. The residues of kasugamycin, oxine-copper and fenaminstrobin were detected using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) combined with QuEChERS pretreatment method and that of prochloraz was monitored using gas chromatography with an electron capture detector (GC-ECD). The results showed washing processing reduced residues by 8.6%-33.9%. Juicing was an effective process to reduce pesticide residues by 65.6%-98.7% with processing factor (PF) of 0.021-0.339. However, high levels of residual prochloraz, oxine-copper and fenaminstrobin (from 5.777 to 51.533mg/kg) were detected in orange essential oils, with PF of 8.720-14.558. Prochloraz was more easily enriched in wet pomace compared with the other three fungicides. When spraying was conducted in accordance with good agricultural practices and using the recommended highest pesticide application dosage and frequency, the dietary intake exposure risks of the four residual fungicides in orange and its processed juices in China were less than 100% for different age groups and caused no unacceptable risks to human health.

Unsaturated Fatty Acid Profiles and Prognostic Significance in Epilepsy Patients: A Comprehensive Analysis Using UPLC-MS/MS and SVM Algorithm

Unsaturated fatty acids (UFAs) play a crucial physiological role in human body. However, the concentration-related changes and prognostic significance of UFAs in epilepsyremain unclear. An optimized ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) approach was developed to measure six key UFAs: oleic acid (OA), linoleic acid (LA), arachidonic acid (AA), α-linolenic acid (ALA), docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA). Subsequently, the levels of these six UFAs were determined in 40 healthy individuals and 49 epilepsy patients. The diagnostic value of UFAs and clinical examination indicators were assessed using statistical analysis and the support vector machine (SVM) algorithm. The results showed that the UPLC-MS/MS method successfully quantified the levels of OA, LA, AA, ALA, EPA, and DHA in both the healthy individuals and epilepsy patients. Compared with the healthy group, the levels of ALA, AA, and DHA were significantly elevated in the epilepsy group (P < 0.05). Pearson correlation analysis revealed a strong positive correlation among the UFAs in the epilepsy group. The orthogonal partial least squares-discriminant analysis (OPLS-DA) model showed that DHA and EPA were more important than cholesterol in distinguishing between two groups, although the separation was not complete. The SVM model achieved better separation, with an area under the curve (AUC) of 0.95 when including the six UFAs. The EPA/DHA ratio was identified as a key feature, with a significant contribution to the model's performance. Removing the six UFAs from the model reduced the AUC to 0.91, highlighting the predictive value of UFAs for epilepsy. In conclusion, ALA, AA, and DHA, are altered in epilepsy patients. The EPA/DHA ratio was found to be a key predictive indicator for epilepsy. The use of UFAs in conjunction with clinical examination data improved the predictive power of the SVM model, suggesting that UFAs have potential as biomarkers for epilepsy.

Volatile organic compounds (VOC) metabolites in urine are associated with increased systemic inflammation levels, and smokers are identified as a vulnerable population

BackgroundPrevious studies indicated that exposure to VOCs was linked to increased systemic inflammation levels. However, the dose-response relationships between urine VOCs metabolites and systemic inflammation have not been established, and the key metabolite of the toxic compounds has not been identified. MethodsWe used data in 7007 US adults in the NHANES cycles (2011–2018) across 8 years. Urinary VOC metabolites were measured using ultra-performance liquid chromatography and electrospray tandem mass spectrometry (UPLC-ESI/MSMS). VOC metabolites were adjusted by urinary creatinine level before analysis. Systemic inflammation was assessed by systemic immune-inflammation index (SII) and systemic inflammation response index (SIRI) indices. Generalized linear models, restricted cubic splines (RCS), and weighted quantile sum (WQS) regression were applied to evaluate the associations, exposure-response (E-R) curve and identify the key contributor compound, adjusting for gender, age, race, BMI, marital condition, education level, smoking level, alcohol consumption and physical activity. Smoking status was assessed as an effect modifier. ResultsSignificant and robust positive correlations were found between 8 VOC metabolites and both SII and SIRI. They were N-Acetyl-S-(2-carboxyethyl)-L-cysteine (CEMA), N-Acetyl-S-(2-cyanoethyl)-L-cysteine (CYMA), N-Acetyl-S-(3,4-dihydroxybutyl)-L-cysteine (DHBMA), N-Acetyl-S-(3-hydroxypropyl)-L-cysteine (3HPMA), mandelic acid (MA), N-Acetyl-S-(4-hydroxy-2-butenyl)-L-cysteine (MHBMA3), phenylglyoxylic acid (PGA), and N-Acetyl-S-(3-hydroxypropyl-1-methyl)-L-cysteine (HPMMA). The RCS curves showed J-shaped or exponential shaped E-R relationships for most VOC metabolites. WQS regression found that exposure to the mixture of VOC metabolites was related to increased systemic inflammation, and MA was the key VOC metabolite contributing most to systemic inflammation levels. Smokers exhibited higher levels of urinary VOCs and larger susceptibility to VOC-related increases in SII and SIRI compared to non-smokers. ConclusionThis study demonstrated a strong link between urinary VOC metabolites and increased systemic inflammation, and smokers were more susceptible. Our findings highlighted the significance of reducing VOC exposure to mitigate the inflammation levels, particularly for smokers.

Central and peripheral kynurenine pathway metabolites in COVID-19: Implications for neurological and immunological responses

Long-term symptoms such as pain, fatigue, and cognitive impairments are commonly observed in individuals affected by coronavirus disease 2019 (COVID-19). Metabolites of the kynurenine pathway have been proposed to account for cognitive impairment in COVID-19 patients.Here, cerebrospinal fluid (CSF) and plasma levels of kynurenine pathway metabolites in 53 COVID-19 patients and 12 non-inflammatory neurological disease controls in Sweden were measured with an ultra-performance liquid chromatography-tandem mass spectrometry system (UPLC-MS/MS) and correlated with immunological markers and neurological markers. Single cell transcriptomic data from a previous study of 130 COVID-19 patients was used to investigate the expression of key genes in the kynurenine pathway.The present study reveals that the neuroactive kynurenine pathway metabolites quinolinic acid (QUIN) and kynurenic acid (KYNA) are increased in CSF in patients with acute COVID-19. In addition, CSF levels of kynurenine, ratio of kynurenine/tryptophan (rKT) and QUIN correlate with neurodegenerative markers. Furthermore, tryptophan is significantly decreased in plasma but not in the CSF. In addition, the kynurenine pathway is strongly activated in the plasma and correlates with the peripheral immunological marker neopterin. Single-cell transcriptomics revealed upregulated gene expressions of the rate-limiting enzyme indoleamine 2,3- dioxygenase1 (IDO1) in CD14+ and CD16+ monocytes that correlated with type II-interferon response exclusively in COVID-19 patients.In summary, our study confirms significant activation of the peripheral kynurenine pathway in patients with acute COVID-19 and, notably, this is the first study to identify elevated levels of kynurenine metabolites in the central nervous system associated with the disease. Our findings suggest that peripheral inflammation, potentially linked to overexpression of IDO1 in monocytes, activates the kynurenine pathway. Increased plasma kynurenine, crossing the blood–brain barrier, serves as a source for elevated brain KYNA and neurotoxic QUIN. We conclude that blocking peripheral-to-central kynurenine transport could be a promising strategy to protect against neurotoxic effects of QUIN in COVID-19 patients.

Determination of sacubitril and seven sartan drugs in serum samples by online solid phase extraction-liquid chromatography-tandem mass spectrometry

An online solid phase extraction–ultra performance liquid chromatography–tandem mass spectrometry detection method was developed for the simultaneous determination of sacubitril and seven sartan drugs in blood serum in this study. The compounds were separated through a C18 column. Mass spectrometry of the samples was performed using a jet stream electrospray ion source (AJS, ESI+). The samples were detected via a multiple reaction monitoring (MRM) mode and quantified via a stable isotope internal standard method. 900 μL of prepared sample was injected and a run time of 12.5 minutes was obtained in this proposed method. The eight examined target compounds showed good linearity (r2＞0.994) in the corresponding mass concentration range and a lower limit of quantitation (LLOQ) was in the range of 0.05–0.1 μg/L. The recovery of the spiked serum samples ranged from 90.90 % to 106.20 % with relative standard deviations (RSDs) of 4.57 %–9.27 %. Five target compounds were detected in the actual serum samples using this method. The proposed method is simple to use, sensitive, accurate, and suitable for the trace detection of sacubitril and seven sartan drugs present in serum samples. The method can meet the needs for clinical monitoring of blood concentrations of this type of drug, while providing detection technology support for the development of compound preparations of sacubitril and other sartan drugs.

Widely Targeted Metabolomics Reveal the Distribution of Metabolites in Shatian Pomelo Fruit.

Using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) technology in multiple reaction monitoring mode, a widely targeted metabolomics approach was employed to identify metabolites in five tissues (exocarp, endocarp, segment membrane, pulp, and seeds) of the Shatian pomelo fruit. The differences in metabolite composition and abundance among different tissues were analyzed using multivariate statistical analysis methods. The results showed that a total of 1722 metabolites were identified from the five tissues of the Shatian pomelo, including 413 flavonoids and 277 amino acids and their derivatives. Flavonoid metabolites accumulate the most abundantly in the exocarp and seeds, while amino acids and their derivatives are primarily accumulated in the exocarp and pulp. A total of 649 key differential metabolites were screened, including flavonoids, amino acids, and their derivatives, indicating the presence of tissue-specific accumulation of metabolites in the Shatian pomelo. This study systematically investigated the metabolite distribution in different tissue parts of the Shatian pomelo, and validated the feasibility of widely targeted metabolomics technology in pomelo quality analysis. It provided a theoretical reference for metabolic research on the Shatian pomelo and other citrus fruits, and offered a theoretical basis for the efficient utilization of pomelo resources.

Enantioseparation, Absolute Configuration, and Enantioselective Bioactivity Mechanism of the Chiral Fungicide Nuarimol.

In this study, the nuarimol enantiomers were successfully baseline separated with Rs 1.70 by ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The absolute configurations of the nuarimol enantiomers were confirmed as R-(+)-nuarimol and S-(-)-nuarimol. The enantioselective bioactivity assay indicated that R-(+)-nuarimol exhibited greater potency against seven phytopathogenic fungi, with values approximately 1.4-3.5 and 4.5-51.4 times higher than those of rac-nuarimol and S-(-)-nuarimol. The active contribution value of R-enantiomer was 82-98%, showing that R-(+)-nuarimol played a crucial role in bioactivity. Meanwhile, R-(+)-nuarimol exhibited stronger effects in increasing the cell membrane permeability, compromising the cell membrane integrity, and inhibiting ergosterol biosynthesis. Molecular docking analysis showed that R-(+)-nuarimol possessed a stronger binding affinity to sterol 14-α demethylase (CYP51) than S-(-)-nuarimol, with docking energies of -7.42 and -7.36 kcal/mol. This study contributes essential data for screening a high-activity enantiomer of nuarimol and provide guidance for reducing used dosage and increasing the efficiency of nuarimolAQ.

Alterations of endocannabinoid signaling and microglia reactivity in the retinas of AD-like mice precede the onset of hippocampal β-amyloid plaques.

Extra-cerebral manifestations of Alzheimer's disease (AD) develop in the retina, which is, therefore, considered a "window to the brain". Recent studies demonstrated the dysregulation of the endocannabinoid (eCB) system (ECS) in AD brain. Here, we explored the possible alterations of ECS and the onset of gliosis in the retina of AD-like mice. Tg2576 (TG) mice overexpressing the amyloid precursor protein (APP) were used at the age of 12 months, when hippocampal β-amyloid plaques had not been developed yet. Analysis of retinal gliosis showed a significant increase in the number of IBA1 (+) microglia cells in TG versus wild type (WT). Gliosis was not associated with retinal β-amyloid plaques, evident retinal degenerative signatures, or excitotoxicity; instead, oxidative stress burden was observed as increased acrolein levels. Analysis of the ECS (receptors/metabolic enzymes) through western blotting (WB) revealed the up-regulation of cannabinoid receptor 2 (CB2) and monoacylglycerol lipase (MAGL), the enzyme responsible for the degradation of 2-arachidonoylglycerol (2-AG), in TG retinas. Fluorescence intensity analysis of anti-CB2 and anti-MAGL immuno-stained cryosections was consistent with WB, showing their up-regulation throughout the retinal layers. No statistically significant differences were found for the other enzymes/receptors of the ECS under study. However, linear regression analysis for individual animals showed a significant correlation between CB2 and fatty acid amide hydrolase (FAAH), diacylglycerol lipase α/β (DAGLα/β), and APP; instead, a significant negative correlation was found between MAGL and APP. Finally, ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) demonstrated a significant reduction of 2-AG in TG retinas (~0.34 ng/mg) compared to WT (~1.70 ng/mg), while a trend toward increase was found for the other eCB anandamide (AEA). Overall, our data indicate that gliosis and ECS dysregulation-in particular of CB2, MAGL and 2-AG-occur in the retina of AD-like mice before retinal degeneration and development of hippocampal β-amyloid plaques.

Detected substance abuse among injecting drug users through analysis of used syringes in Tunisia

Background and aimsInjecting drug use poses significant public health risks due to unsafe practices such as syringe sharing, reuse, and risky sexual behaviors, which increase the transmission of bloodborne viruses. In Tunisia, limited data on injecting drug use hinders the development of informed health and harm reduction policies. MethodsA syringe collection campaign was conducted in Tunis in November 2022. The used syringes were provided by the Tunisian Association for Information and Orientation on AIDS and Addiction (ATIOST), a harm-reduction service. These syringes had been distributed to people who inject drugs (PWID) as part of a mobile syringe exchange program. The objective of the study was to analyze the contents of the used syringes to gain further insights into drug use patterns among PWID. The residual substances in the syringes were examined using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), in accordance with the standardized protocol of the European Syringe Collection and Analysis Project Enterprise (ESCAPE). ResultsA total of 261 syringes from five collection sites were analyzed. Among these, 87 % contained at least one psychoactive substance, while 32 % contained more than two psychoactive substances. The most frequently identified psychoactive substances were buprenorphine (50.28 %), amphetamine (11.65 %) and tramadol (9.66 %). No substances were detected in 34 syringes. ConclusionThis method provides rapid data on drug use trends in specific regions and timeframes, revealing differences that can inform tailored prevention and harm reduction strategies. Such analyses are valuable for comparative studies across countries in the European Neighbourhood Policy (ENP-South) region.

Astragalus polysaccharide alleviates asthma by modulating gut microbiota and serum metabolomics.

The aim of the present study was to examine the effects of Astragalus polysaccharide (APS) on gut microbiota and serum metabolites in asthmatic mice. For this purpose, a total of 36 BALB/c female mice were selected and randomly classified into the following groups: i) The normal control group sensitized with phosphate-buffered saline; ii) the asthma group sensitized and challenged with ovalbumin (OVA); iii) the OVA + APS (2.5 g/kg) treatment group; iv) the OVA + APS (5.0 g/kg) treatment group; v) the OVA + APS (10 g/kg) treatment group; and vi) the OVA + dexamethasone (2 mg/kg) treatment group, with 6 mice in each group. OVA was used to establish the mouse model of asthma. In the APS group, the asthmatic mice were intragastrically administered APS at various doses at 1 h prior to each OVA stimulation. The airway hyperreactivity (AHR) was measured, and hematoxylin and eosin staining was employed to evaluate pulmonary inflammatory infiltration. In addition, 16S rRNA sequencing and ultra-performance liquid chromatography-tandem mass spectrometry were used to detect the changes in the mouse gut microbiota and serum metabolites. The results revealed that compared with the asthma model group, APS improved airway inflammation and eosinophil infiltration in asthmatic mice. In asthmatic mice, the gut microbial imbalance mainly manifested as a low abundance of Bacteroidetes and a high abundance of Firmicutes, yielding an increased F/B ratio. In the high-dose APS group, the abundance of Firmicutes was reduced, and the abundance of Bacteroidetes was increased, which thereby decreased the F/B ratio and corrected the gut microbial imbalance. Through blood metabolomics, 145 and 105 significantly differential metabolites were detected in the medium- and high-dose APS groups, respectively. Moreover, Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis demonstrated that the metabolic pathways in the medium-dose APS group included the biosynthesis of unsaturated fatty acids and the biosynthesis of arginine. On the other hand, the metabolic pathways enriched in high-dose APS group were the biosynthesis of unsaturated fatty acids, and pyrimidine metabolism. On the whole, the present study demonstrates that APS may regulate the gut microbiota and the metabolites to improve airway inflammation and AHR in asthmatic mice.

Effect of dietary supplementation of yeast culture Saccharomyces cerevisiae in lactating female goats.

This study was designed to investigate the effects of adding a novel yeast culture, Saccharomyces cerevisiae refermented sorghum distiller's dried grains with solubles (SSDDGS), to the diets of lactating female goats on lactation performance and lamb growth performance. We divided 10 lactating Dazu black goats of similar age, weight, and offspring into two groups: one fed a pelleted diet with 50 g/day SSDDGS (ET), and the other without SSDDGS as a control (EC) for 7 weeks. We monitor the weight changes of each goat and collect blood and milk samples from experimental ewes at specific times for hormone and milk composition determination. We use ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) to detect metabolites in the serum of lactating ewes. Our results showed that SSDDGS supplementation significantly reduced female goats' average daily weight loss during weeks 2-4 of lactation and increased serum IGF-1 and prolactin levels at week 4 (p < 0.05). SSDDGS supplementation in early lactation significantly increased milk protein, lactose, and ash content (p < 0.05). UPLC-MS/MS analysis showed that SSDDGS changed the levels of 58 metabolites in the serum of lactating goats. These metabolites were mainly involved in the sohingolipid signaling pathway, and cysteine, methionine, and sphingolipid metabolism. In summary, Yeast culture SSDDGS reduced weight loss, enhanced milk quality, and modified metabolic profiles in early lactation goats, providing insight into the potential regulatory role and mechanism of yeast culture in lactation female goats.

Integrative Transcriptomic and Metabolomic Analysis Reveals Quinoa Leaf Response Mechanisms to Different Phosphorus Concentrations During Filling Stage

Quinoa is an annual self-pollinating plant rich in polyphenols, flavonoids, saponins, and amino acids; its protein balance closely aligns with the ideal recommendation set by the Food and Agriculture Organization. Therefore, quinoa is considered the most suitable “all-nutrient food”. Phosphorus fertilization plays an important role in restricting the growth and development of quinoa; however, the effects of phosphorus fertilizer on quinoa growth remain unclear. Therefore, we conducted metabolome and transcriptome analyses on quinoa leaves during the filling stage, subjecting plants to different doses of phosphorus fertilizer. Overall, phosphorus treatment exerted a significant impact on the phenotypic characteristics of quinoa. Specifically, through a combined analysis of ultra-performance liquid chromatography–tandem mass spectrometry and transcriptome analysis, we identified the alteration and regulation of specific metabolites and genes within flavonoid biosynthesis pathways; this comprehensive evaluation helped elucidate the response mechanism of quinoa leaves during the grouting stage under various phosphorus conditions. Ultimately, the results of this study provide a reference for the selection of quinoa cultivars that exhibit tolerance to low- or high-phosphorus stress; additionally, we offer a theoretical basis for the rational application of phosphorus fertilizer and the enhancement of phosphorus utilization efficiency.

Ergot alkaloids: comparison of extraction efficiencies for their monitoring in several cereal-solvent combinations by UPLC-MS/MS.

To date, there are more than 80 ergot alkaloids identified; their distribution depends on different factors (e.g. geographic regions, host plants). These toxins can cause acute and chronic toxic effects on human health and commonly infect cereal crops such as triticale and rye, wheat, barley and oats. Considering the growing consumption of plant-based foods, the European Food Safety Authority has highlighted the need to develop risk assessment strategies. This work focused on the optimization of extraction efficiency, to quantify the main ergot alkaloids and their epimers, that are available on the market without any legal restriction (ergosine, ergocristine, ergocriptyne, ergocornine, ergosinine, ergocristinine, ergocriptinine and ergocorninine). Considering the quantification of 8 out of 12 regulated compounds by EU (sum of -ine and -inine forms), this approach can be defined as a screening method for a reliable estimation of the risk, specifically devoted to industrial stakeholders that can then possibly outsource to authorized external labs only the samples suspected of significant positivity. The effectiveness of three different extraction conditions (acidic, alkaline and neutral) followed by a rapid clean-up using dispersive solid-phase extraction with C18 sorbent was evaluated by ultra-performance liquid chromatography tandem quadrupole mass spectrometry (UPLC-MS/MS), resulting in a short chromatographic run (16min). The method was developed and validated in five different cereal production chains (rye, oat, wheat, wheat gluten and baby food). The applicability of the method was examined by analyzing a set of 54 samples, including also other cereals like spelt, tritordeum and triticale, and evaluating also some reference materials.

Spatial-temporal Distribution and Ecological Risk of Antibiotics in the Soils of a Typical Agricultural Watershed in the Three Gorges Reservoir Area

In order to understand the spatial-temporal distribution and ecological risk of antibiotics in the soil of an agricultural watershed in the Three Gorges Reservoir area, the topsoil samples were collected at 26 sites in the Wangjiagou small watershed, Fuling District, Chongqing in spring, summer, autumn, and winter of 2022, and 21 antibiotics with five classes were determined using solid phase extraction and ultra-performance liquid chromatography-tandem mass spectrometry. The content levels and spatial-temporal distribution of antibiotics were analyzed, the correlations between antibiotic contents and soil physicochemical factors were discussed, and the potential ecological risk of antibiotics in the soil was evaluated using the risk quotient method. The results showed that the detection rates of 21 antibiotics were 0-100% with the range of ND-219.5 μg·kg-1, and those of tetracyclines （TCs）, quinolones （FQs）, and chloramphenicols （CAPs） reached 100% in all four seasons. The total antibiotic content ranged from 14.35 to 504.1 μg·kg-1 with the average value of 149.7 μg·kg-1, and the average contents of the five classes of antibiotics showed a decreasing trend of TCs （77.95 μg·kg-1）, FQs （34.96 μg·kg-1）, CAPs （28.14 μg·kg-1）, sulfonamides （SAs, 7.15 μg·kg-1）, and macrolides （MLs, 1.48 μg·kg-1）. The antibiotic contents of soils during autumn and winter were significantly higher than those in summer （P &lt; 0.05） and showed an overall variation trend of first decreasing and then increasing with the season and a distribution situation of "low in the west and high in the east." The sites with high antibiotic contents were mainly concentrated in vegetable fields, residential areas, and dry lands. In spring soils, the contents of tylosin （TYL） （P &lt; 0.01）, tetracycline （TC） （P &lt; 0.01）, doxycycline （DXC）, and sulfamethodiazine （SMR） （P &lt; 0.05） were significantly negatively correlated with soil pH. In summer soils, total phosphorus （TP） had significantly positive effects on the contents of sulfamethoxazole （SFZ） and TC （P &lt; 0.05） and had extremely significant positive effects on sulfamethoxazole （SMX）, SMR, and ofloxacin （OFL） （P &lt; 0.01）. Total nitrogen （TN） had significantly positive effects on the contents of SMX （P &lt; 0.01） and SMR （P &lt; 0.05）, and soil organic matter （SOM） had significantly positive effects on the contents of TC and enrofloxacin （ENR） （P &lt; 0.05）. However, ciprofloxacin （CIP） （P &lt; 0.001）, sulfampyridine （SPD）, norfloxacin （NOR）, and TYL contents （P &lt; 0.05） were significantly negatively correlated with soil pH. In autumn soils, TN significantly negatively affected CIP content （P &lt; 0.05）； the contents of sulfadiazine （SDZ）, TC, and DXC were negatively affected by soil pH （P &lt; 0.05）； and cation exchange capacity （CEC） significantly positively affected the contents of TC and SFZ （P &lt; 0.05）. In winter soils, TP had a significantly positive effect on TYL content, and TN had a significantly positive effect on SFZ content （P &lt; 0.05） and an extremely positive effect on sulfamethoxine （SMM） （P &lt; 0.001）. Chlorotetracycline （CTC） （P &lt; 0.01）, CIP （P &lt; 0.01）, thiamphenicol （TAP） （P &lt; 0.001）, chloramphenicol （CAP） （P &lt; 0.001）, oxytetracycline （OTC） （P &lt; 0.001）, and sulfadimidine （SM2） （P &lt; 0.05） were significantly negatively correlated with soil pH, and CEC had a significantly negative effect on SDZ content （P &lt; 0.05）. The soils in the Wangjiagou agricultural small watershed were mainly threatened by SMX, SDZ, TC, DXC, erythromycin, NOR, OFL, CIP, and ENR, and their ecological risks should not be ignored.

Adenosine regulates depressive behavior in mice with chronic social defeat stress through gut microbiota

Major depressive disorder (MDD) is recognized as the most prevalent affective disorder worldwide. Metagenomic studies increasingly support a critical role for dysbiosis of gut microbiota in the development of depression. Previous studies have demonstrated that adenosine alleviates gut dysbiosis, suggesting that elevating adenosine levels could be a novel intervention for MDD; however, the mechanisms underlying this effect remain unclear. This study utilized 16S rRNA gene sequencing, fecal microbiota transplantation (FMT) and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to test the hypothesis that increased adenosine alleviates depressive behaviors in male mice subjected to chronic social defeat stress (CSDS) through alterations to gut microbiota. The data showed that depression-susceptible (SUS) mice exhibited gut dysbiosis, and FMT from SUS mice increased depression-like behaviors in healthy recipients. In SUS mice, adenosine supplementation ameliorated both depression-like behaviors and abnormalities in gut microbiota, and co-administration of probiotics and adenosine not only mitigated depression-like behaviors but also enhanced gut barrier integrity. By including 83 depressed adolescents and 67 healthy controls, this study found that the level of short-chain fatty acids (SCFAs) in the depression group was reduced, this finding parallels reductions seen in SUS mice and in recipient mice after FMT from SUS donors. Conversely, supplementation with either adenosine or probiotics led increased SCFAs concentrations in the serum of SUS mice. These findings suggest that adenosine may alleviate depression-like behaviors in CSDS mice by modulating the gut microbiota. This effect is likely associated with increased serum SCFAs, metabolites produced by the gut microbiota, following adenosine supplementation.

Evaluation of protein impurities in Ademetionine 1,4-Butanedisulfonate

Ademetionine 1,4-Butanedisulfonate (SAMe) is widely used as a prescription drug to treat cholestasis associated with liver disease, and is also used to treat depression, Alzheimer's disease and other diseases. Currently, the main way to produce SAMe is by fermentation of S-adenosylmethionine synthetase（SAMS）. However, during the fermentation process, host cells produce contaminants unrelated to the treatment. Among them, host cell protein (HCP), which is co-purified with the drug, is the main impurity in the production process. HCP-induced antigenic reactions can lead to allergies or other adverse reactions and affect drug quality, efficacy, and safety. Enzyme-linked immunosorbent assay (ELISA) and mass spectrometry (MS) are currently used as the main analytical methods for measuring HCP. The ELISA method may ignore some HCPs with low immunogenicity, while mass spectrometry is primarily employed for the characterization of proteomes. Orthogonal methods can more effectively evaluate impurities in drugs. In this study, ELISA was initially utilized to quantify the content of Saccharomyces cerevisiae host protein in 11 batches of SAMe active pharmaceutical ingredient (API) from two suppliers, then the above APIs were qualitatively analyzed by MS method. Following the identification of surrogate peptide of SAMS, an Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry（UPLC-MS/MS）method was established and validated, the SAMS protein residues in 11 batches of samples were subsequently determined. The results demonstrated that the levels of Saccharomyces cerevisiae host protein and SAMS in the samples were both low, with values below 20 ppm and 1.19 ppm, respectively. The qualitative results also indicate that the types of peptide residues in the samples are more diverse than those of protein residues. Furthermore, residues of proteins and peptides can be detected in all samples, which underscores the importance of evaluating the HCP in API. This study employs a range of complementary detection methods to conduct a comprehensive and sensitive evaluation of total HCP and specific proteins in drugs, thereby guiding more informed assessments of the risks posed by HCP impurities in raw materials during pharmaceutical processes.