Biomedical Chromatography最新文献

A sensitive and robust LC-MS/MS method was developed and validated for the quantification of vibegron in human plasma, employing electrospray ionization in positive mode and multiple reaction monitoring (MRM). The precursor ion m/z 445.5 yielded two product ions: m/z 427.4 and m/z 135.1. Despite higher intensity, m/z 427.4 was excluded due to chromatographic interference, while m/z 135.1, previously unreported for vibegron, was selected for its superior selectivity and specificity. A stable isotope-labeled internal standard, vibegron ¹³C₅ (m/z 450.3 → 139.1), was used to enhance precision and minimize matrix effects. Chromatographic separation was achieved on a Kromasil C18 column with a mobile phase of acetonitrile and 10-mM ammonium formate (pH 5.8), yielding a retention time of 1.33 min and total runtime of 3.00 min. Liquid–liquid extraction with ethyl acetate provided consistent recovery (87.7%), and sodium fluoride was added to inhibit enzymatic degradation. The method demonstrated excellent linearity (0.500–400.223 ng/mL), precision (0.58%–3.34%), and accuracy (98.26%–102.29%), using only 100 μL of plasma. Fully compliant with ICH M10 guidelines, the method is well suited for future pharmacokinetic, bioavailability, and bioequivalence studies.

Celastrus orbiculatus fruits as Beihehuan (BHH) in Chinese are used as an alternative to Albizia julibrissin flowers (Hehuanhua) for insomnia treatment in North of China. Both of them perhaps contain similar material foundations with common components, making them highly worthy of systematic analysis of the ingredients and related pharmacological mechanisms. In this study, the components in BHH were comprehensively analyzed by LC–MS/MS-based molecular networking technology combined with literature reports, and important targets and core active ingredients in BHH that can treat insomnia were screened through network pharmacology and molecular docking analysis. Finally, a total of 136 components were annotated and summarized. BHH had flavonoids, fatty acids, and volatile oil as similar or the same component categories and structures as Hehuanhua, especially different categories of dihydro-β-agarofuran-type sesquiterpenoids with similar functions. The screened top 10 KEGG pathways involved signal transduction related to neurotransmitters, hormones, and addictive substances, and the selected effective ingredients could play synergistic roles in inflammation regulation and neuroprotection. These results provide that BHH may be able to replace Hehuanhua in treating insomnia and other neurological disorders; modern research can provide the scientific connotation and basis for the proposed forward-thinking TCM theory of “effect-oriented source unification” and “heteroagent possessing homofunction.”

Elacestrant is an orally administered selective estrogen receptor degrader designed for the treatment of ER-positive, HER2-negative breast cancer. In this study, a simple and sensitive HPLC–MS/MS method was developed and validated for the quantification of elacestrant in liver microsomes. Chromatographic separation was achieved on a Waters ACQUITY BEH C₁₈ column using a gradient of 0.1% formic acid in water and acetonitrile. Elacestrant and the internal standard (elacestrant-d₆) were detected in multiple reaction monitoring mode via the transitions of m/z 459.2 → 298.3 and m/z 465.3 → 304.4, respectively. The method exhibited excellent linearity over the range of 1.0–2000 nM (r > 0.995). Elacestrant was rapidly metabolized, showing half-lives of 22.45 ± 0.66 min in rat liver microsomes and 43.36 ± 2.48 min in human liver microsomes. Using HPLC–Q-Orbitrap–HRMS, seven metabolites were identified, with M5 (N-deethylation) being the most abundant. Key metabolic pathways involved O-demethylation, N-deethylation, N-dealkylation, and oxidative deamination, primarily mediated by cytochrome P450 3A4. This study establishes the first HPLC–MS/MS and HPLC–Q-Orbitrap–HRMS–based analytical strategy for in vitro metabolic profiling of elacestrant, supporting its future application in clinical pharmacokinetic and metabolism investigations.

A highly sensitive and reliable ultrahigh performance liquid chromatography–tandem mass spectrometry method (UHPLC-MS/MS) has been developed for the quantification of saxagliptin (SAX) and its metabolite, 5-hydroxy saxagliptin (5-OH-SAX), in human plasma. Utilizing an ACE Excel 3 Super C18 column with electrospray ionization multiple reaction monitoring, this method achieves excellent linearity across the concentration ranges of 0.1–50 ng/mL for SAX and 0.2–100 ng/mL for 5-OH-SAX. The mean extraction recoveries are reported at 99.87% for SAX and 97.68% for 5-OH-SAX, with matrix effects maintained at 0.85 and 0.86, respectively. Both intraday and interday relative standard deviations are maintained below 15%. Furthermore, the developed method confirms appropriate short-term stability, satisfying the requirements for pharmaceutical analysis without residual effects. Significantly, this validated method applies to pharmacokinetic studies, providing crucial insights into the drug and metabolite behaviors during varying ingestion states, thus enhancing therapeutic efficacy and patient compliance through informed SAX administration.

A rapid and reliable liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was established for the quantification of contezolid in human plasma and liver tissue. Sample preparation was performed by protein precipitation using linezolid as the internal standard. Chromatographic separation was achieved on an ACQUITY BEH C18 column (2.1 × 50 mm, 1.8 μm) with a mobile phase consisting of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B), delivered at a flow rate of 0.5 mL/min. Quantification was performed in multiple reaction monitoring (MRM) mode, monitoring the transitions m/z 409.1 → 269.1 for contezolid and m/z 338.1 → 296.2 for linezolid. The method showed excellent linearity over the ranges of 20–10,000 ng/mL in plasma and 100–50,000 ng/g in liver tissue. Full validation confirmed that the method met regulatory standards for accuracy, precision, sensitivity, selectivity, stability, matrix effects, carryover, and interference. The validated assay was successfully applied to a pharmacokinetic study of contezolid in liver transplant recipients. In conclusion, this robust, efficient, and cost-effective LC–MS/MS method provides a valuable tool for the determination of contezolid in clinical and pharmacokinetic research.

Salvia species are valuable natural resources due to their rich essential oils and bioactive compounds with applications in medicine, food, and cosmetics. In this study, essential oils and aroma components of S. montbretii and S. hydrangea were analyzed using GC–MS/FID and headspace GC–MS. Terpenoids were identified by GC–MS, while phenolic and flavonoid compounds were quantified by LC–MS/MS. Total phenolic and flavonoid contents, antioxidant capacities, and enzyme inhibitory activities were also evaluated. Caryophyllene was the dominant compound in S. montbretii essential oil and aroma samples, and camphor in S. hydrangea. GC–MS results indicated high levels of ursolic (54,675.22–1583.23 μg/g) and oleanolic acids (100,567.85–2592.11 μg/g), while LC–MS/MS revealed abundant rosmarinic acid (up to 27,0857–587.85 μg/g). All extracts exhibited moderate to strong antioxidant activities and notable inhibitory effects against urease and cholinesterase. Cytotoxic effects were observed on colon (Caco-2) and ovarian (SkOV3) cancer cell lines, along with toxicity in healthy cells (PDF). In conclusion, S. montbretii and S. hydrangea may serve as potential natural sources for pharmaceutical and cosmetic applications; however, detailed safety and dosage assessments are required prior to practical use due to the observed cytotoxic effects.

Apalutamide (Erleada) is an oral androgen receptor antagonist that inhibits androgen action, reducing cancer cell growth. The FDA approved apalutamide (APD) for metastatic castration-sensitive prostate cancer (mCSPC) and nonmetastatic castration-resistant prostate cancer. This study created a fast, reliable, and sensitive UPLC-MS/MS method to assess APD metabolic stability in human liver microsomes (HLMs). The greenness of the method was assessed using in silico applications like AGREE and MoGAPI. The StarDrop package (WhichP450 and DEREK modules) identified APD chemical structure warnings and assessed metabolic lability. An isocratic mobile phase system with a C18 column segregated APD and baricitinib (BCB as internal standard). APD calibration curve sensitivity was high, with statistical linearity from 1.0 to 4000 ng/mL. Intraday and interday evaluation accuracy and precision were 0.72%–10.25% and −0.69% to 12.25%, respectively. The UPLC-MS/MS method used a little amount of acetonitrile (35%), a low flow rate of 0.3 mL/min, and a short running period of 1 min, demonstrating environmental sustainability. APD's in vitro half-life (t_1/2) and intrinsic clearance (Cl_int) were 25.57 min and 31.7 mL/min/kg, respectively. These in silico data propose that APD derivatives with changes in N-methyl moiety might reveal altered metabolic lability; this requires experimental verification.

This study investigated the phytochemical composition and pharmacological activities of Fagonia indica crude methanolic extract (MEFFI) and its aqueous fraction (AQFFI). Phytochemical profiling via gas chromatography–mass spectrometry (GC–MS) identified 19 compounds in MEFFI and 11 in AQFFI. Predominant constituents in MEFFI included n-hexadecanoic acid, octadecanoic acid, phytol, and 2-butoxyethanol, while AQFFI was rich in 4-O-methylmannose, thiocyanic acid ethyl ester, and thiophene derivatives—compounds associated with diverse biological activities, including modulation of intestinal motility. Pharmacological evaluation revealed dose-dependent inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), indicating neuroprotective potential. In antioxidant assays, MEFFI exhibited stronger radical scavenging activity (IC₅₀ = 222 for DPPH and 230 μg/mL for ABTS) compared to AQFFI. Acute toxicity assessment in pigeons confirmed safety of both extracts at doses up to 2 g/kg. In vivo experiments revealed that AQFFI significantly enhanced gastrointestinal transit (83.87%), demonstrating prokinetic activity, whereas MEFFI exhibited pronounced laxative effects in constipation models. Both extracts also produced dose-dependent spasmolytic effects on isolated rabbit jejunum. These findings highlight the therapeutic potential of F. indica extracts in managing gastrointestinal motility disorders and support further mechanistic studies to correlate the specific phytochemicals with their observed laxative and spasmolytic activities.