Biomedical Chromatography最新文献

Prunus mume (PM), the dried flower bud of a Rosaceae plant, has a long history of use for its liver-soothing, depression-relieving, and appetite-stimulating effects. Recently, PM has gained attention for its anti-inflammatory, antioxidant, and uric acid-lowering properties. The chemical composition of PM was analyzed using network pharmacology and liquid chromatography–mass spectrometry (LC-MS). The therapeutic potential of PM for hyperuricemia-induced kidney damage was evaluated in a quail model. Antioxidant activity in an HK-2 cell model of hyperuricemia was assessed by measuring the levels of MDA, SOD, and GSH. Additionally, the anti-inflammatory potential was examined using ELISA to measure TNF-α and IL-6 levels. Western blotting was employed to study the effects on URAT1, GLUT9, and the PI3K/AKT pathway. LC-MS identified 284 compounds in PM, with 35 predicted active ingredients. The quail model demonstrated PM's protective effects on the kidneys under hyperuricemic conditions. In vitro, PM reduced oxidative stress and lowered TNF-α and IL-6 levels. It also modulated URAT1 and GLUT9 expression and influenced the PI3K/AKT pathway. PM shows promise in protecting kidneys from hyperuricemia-induced damage, likely through its anti-inflammatory and antioxidant activities, as well as the regulation of urate transport proteins and the PI3K/AKT pathway.

Idiopathic central precocious puberty (ICPP) is an endocrine disorder increasingly observed in children, commonly treated with gonadotropin-releasing hormone agonists (GnRHa). However, the serum lipid profiles in girls with ICPP and the potential regulatory effects of GnRHa in lipid metabolism remain unclear. This research analyzed lipidomic profiles of serum samples from girls younger than 10 years of age, including 37 patients with ICPP, 32 ICPP patients treated with GnRHa, and 30 age-matched healthy controls. Using UHPLC-Q-Exactive Orbitrap/MS, we identified a total of 898 differential lipids in both positive and negative modes, covering classes such as phosphatidylcholine (PC, including EtherPC), acylcarnitine (CAR), triglyceride (TG), N-acyl ethanolamines (NAEs), diacylglycerol (DG), sphingomyelin (SM), phosphatidylethanolamine (PE), ceramide (Cer), and fatty acids (FAs). In the ICPP cohort, 105 lipids exhibited significant differences compared with controls, with increased levels of CAR, SM, PC, PE, TG, and FA (p < 0.05), and notably decreased NAE levels (p < 0.05). Specifically, NAE 20:1 (AUC: 1.0), NAE 18:2 (AUC: 0.948), and NAE 20:0 (AUC: 0.895) were identified as potential diagnostic biomarkers for ICPP. Following GnRHa treatment, serum peak LH and FSH levels decreased, alongside reversible changes in 44 lipids, predominantly CAR, NAE, and TG. In conclusion, this study demonstrates that ICPP is associated with significant alterations in lipid metabolism. The lipidomic changes induced by GnRHa therapy suggest a potential link between ICPP treatment and lipid homeostasis, which warrants further investigation to refine therapeutic approaches for ICPP and potentially mitigate associated metabolic disorders.

As a traditional Chinese medicine, velvet antler (VA) is usually processed with white wine according to the Chinese Pharmacopoeia. The practice of using aged vinegar to process VA is uncommon, which is only used in “GuiLingJi.” In this study, we found significant chemical changes in vinegar processing. Network pharmacology analysis showed that 20 increased components mainly were related with fecundity through the regulation of biosynthesis of steroid hormones and the estrogen signaling pathway. Reproduction experiment using Drosophila melanogaster showed that both VA and vinegar-processed velvet antler (VPVA) could enhance the reproductive capacity and increase the steroid hormone levels in Drosophila, while VPVA was much superior to VA. In addition, metabolomics showed that energy metabolism was related with the mechanisms by which VA improves the fecundity of Drosophila. This study provides a theoretical basis for the rationale of vinegar processing of VA.

Apixaban is an oral anticoagulant classified as a direct factor Xa inhibitor. It is widely utilized for prophylaxis and management of thromboembolic conditions, including stroke, deep vein thrombosis, and pulmonary embolism, as a primary therapeutic option. Apixaban is subjected to stringent testing for potential genotoxic impurities during its manufacturing, due to its structure indicating the possibility of such impurity's formation. Setting limits in accordance with ICH M7 guidelines is essential to reduce patient exposure and also recommended an advanced analytical tool for testing. The present study aims to establish limits based on computational toxicological evaluation, and develop and validate a highly sensitive LS-MS/MS method for quantifying three novel genotoxic impurities of apixaban, namely impurities F, G, and H. The method was established utilizing a mobile phase comprising a pH 5.5 acetate buffer and acetonitrile in a gradient mode. Used a C18 column, of 150 mm (length) × 3.0 mm (width), 2.7 μm of particle size as stationary phase. Quantification was performed with multi-response monitoring in mass spectrometry, employing precursor ions of m/z 437.2, 525.2, and 569.1 for impurities F, G, and H, respectively. The established method is validated for its intended use in accordance with regulatory guidelines and found suitable.

Niraparib is a small-molecule inhibitor of poly(adenosine diphosphate [ADP]-ribose) polymerase 1/2, which is used to treat ovarian cancer. Elevated maximum blood concentrations of niraparib and the area under the blood concentration–time curve (AUC) were correlated with body weight up to 77 kg. Lower body weight increases blood niraparib concentrations and the AUC of ovarian cancer in Asian patients. Therefore, therapeutic drug monitoring (TDM) of ovarian cancer drugs may increase niraparib efficacy and minimize adverse events. In this study, we quantified niraparib in human plasma (50 μL) using a simple and specific HPLC–UV method. The analyte was separated on a reversed-phase column with an isocratic mobile phase of 0.5% KH₂PO₄ (pH 4.5) and acetonitrile (75:25, v/v) at a flow rate of 1.0 mL/min. Calibration curves were linear over 0.25–5 μg/mL (r² = 0.9998). Intraday and interday precision ranged from 2.25% to 6.29% and 1.73% to 3.20%, respectively, whereas accuracy and recovery ranged from −6.02% to −1.75% and > 93.2%, respectively. We cost-effectively quantified steady-state niraparib concentrations in the plasma of patients with ovarian cancer. Therefore, our method could be applied to the departments of pharmacy and clinical laboratories in general hospitals to facilitate the TDM of niraparib without the need for LC–MS/MS.

Alzheimer's disease (AD) is a common neurodegenerative disease for which there are no effective drugs. Kai-Xin-San (KXS), with definite curative effects, is widely used for the prevention and treatment of AD in China. But its mechanism is not yet fully understood. Based on our established rat model and previous pharmacodynamics study, Multi-omics (metabolomics, proteomics) and network analysis were integrated to explore the holistic mechanism of anti-AD effects of KXS. The key pathways were validated with western blot and ELISA methods. Morris water maze and Nissl staining showed that KXS could ameliorate cognitive deficits and pathological morphology of the hippocampus in AD rats. A total of nine metabolites were identified, which were related to pyrimidine metabolism, riboflavin metabolism, tyrosine metabolism, tryptophan metabolism, and glycerophospholipid metabolism. Proteomics results indicated that the improvement of cognitive deficits by KXS was closely related to the regulation of oxidative phosphorylation in mitochondria. Western blotting results showed that KXS significantly inhibited the expression of Mt-nd2 and Ndufb6 in AD rats. Integrated analysis indicated that the anti-AD targets of KXS were interrelated and KXS could exert its anti-AD effect by reducing oxidative stress, neurotoxicity, and inflammation.

Kaixin San (KXS) is a classical prescription for the treatment of depression. However, the mechanism is not clear. In this study, serum pharmacochemistry, mediated by the UHPLC-Orbitrap Exploris 480 mass spectrometer, was used to identify compounds derived from the KXS-medicated serum. These components were used to construct a compound-target network for depression using a network pharmacology approach to predict potential biological targets of KXS. Subsequently, we established a mouse model of CUMS-induced depression and observed the antidepressant effect of KXS. The signalling pathways predicted by the network pharmacology were further validated in animal experiments. The results showed that 36 compounds were identified from the KXS-medicated serum. Based on this, 984 genes related to the compounds and 4966 genes related to depression were identified using network pharmacology. Critically, KEGG analysis identified the PI3K/Akt and NF-κB signalling pathways as the main pathways through which KXS exerts its antidepressant effect. KXS significantly alleviated depression-like behaviour and hippocampal histopathological changes in a mouse model of depression. Compared with the model group, the treatment of KXS significantly reduced the expression of protein targets in the PI3K/Akt/NF-κB signalling pathway. All these studies effectively corroborated the predicted results, confirming the feasibility of this integrated strategy.

Bile-processed Coptidis Rhizoma (BPCR) exhibits stronger efficacy in treating T2DM than Coptidis Rhizoma(CR) alone. However, the synergistic mechanism of its processing remains unknown. This study utilized HPLC to determine the content and dissolution characteristics of alkaloid components in BPCR before and after processing. The results indicated that the diffusion of the alkaloids in BPCR is stronger than that of CR, and their dissolution conforms to the Weibull equation. Additionally, BPCR significantly reduced fasting blood glucose (FBG) and serum insulin (FINS) levels in T2DM rats induced by a high-fat diet (HFD) and streptozotocin (STZ), improved glucose and lipid metabolism, and mitigated liver damage. Serum metabolomics analysis based on UPLC-Q-TOF-MS revealed that BPCR significantly regulates 27 endogenous differential biomarkers. The underlying mechanism may be related to glycerophospholipid metabolism, linoleic acid metabolism, steroid biosynthesis, and arachidonic acid metabolism pathways.

A simple and sensitive liquid chromatography tandem mass spectrometry method was established and validated for determination of HRO761 in rat plasma. After prepared by protein precipitation with acetonitrile, HRO761 and internal standard were separated on a Waters BEH C18 column using acetonitrile containing 0.1% formic acid and 0.1% formic acid in water as mobile phase by gradient elution. The method showed excellent linearity over the range of 5–5000 ng/mL with acceptable intraday and interday precision, accuracy, matrix effect, and recovery. The stability assay indicated that HRO761 was stable during the sample acquisition, preparation, and storage. The method was applied to pharmacokinetic study of HRO761 in rats. The result suggested that after intravenous administration at dose of 1 mg/kg, HRO761 was quickly eliminated from the plasma with the elimination half-life of 1.9 h. After oral administration at doses of 5, 10, and 20 mg/kg, HRO761 was quickly absorbed into plasma and reach the peak concentration (C_max) of 2598.1–9379.2 ng/mL at 1.0–4.0 h. The exposure increased proportionally with the dose. The oral bioavailability was 79.0%–99.1% over the range of 5–20 mg/kg. This study provides useful information for its further development in clinic.

The Tibetan medicine Wuwei Shaji powder (WWSJ) has been extensively applied to treating lung diseases such as chronic obstructive pulmonary disease (COPD). However, there are significant limitations in its quality control and evaluation. The purpose of this study was to analyze the overall chemical composition of WWSJ and to predict potential quality markers for WWSJ in the treatment of COPD. Using UHPLC-Q-Exactive Orbitrap-MS, 83 chemical constituents of WWSJ were identified or preliminarily characterized by 14 authentic reference standards, accurate mass counts, and characteristic fragment ions. In addition, serum pharmacology methods were employed to detect 13 blood-migrating components. Further network pharmacological research found that the potential mechanism of WWSJ in the treatment of COPD may be related to the regulation of the immune system and inflammatory response and identified 10 chemicals as key molecules in the treatment of COPD. Finally, the contents of six potential quality markers were determined simultaneously by high-performance liquid chromatography (HPLC). This study lays the foundation for further research on the mechanism of WWSJ in the treatment of COPD.