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Abstract This study aimed to elucidate the clinical diagnostic value of plasma catecholamines and their metabolites for pheochromocytoma and paraganglioma (PPGL)‐induced secondary hypertension using ultraperformance liquid chromatography‐mass spectrometry (UPLC‐MS/MS). The study population included 155 patients with PPGL that were divided into the PPGL with hypertension (n = 79) and a PPGL without hypertension (n = 76) groups, and 90 healthy volunteers and 90 patients with primary hypertension as the control groups. UPLC‐MS/MS was performed to detect plasma levels of catecholamines and their metabolites, including dopamine, vanillylmandelic acid (VMA), norepinephrine, metanephrine, and normetanephrine. Receiver operating characteristic curves were generated to analyze the diagnostic value of the plasma levels of catecholamines and their metabolites in PPGL‐induced secondary hypertension. Patients in the primary hypertension and PPGL without hypertension groups had higher levels of dopamine, VMA, norepinephrine, metanephrine, and normetanephrine than patients in the normal group (all p

Abstract Autism spectrum disorder (ASD) presents a significant risk to human well‐being and has emerged as a worldwide public health concern. Twenty‐eight children with ASD and 33 healthy children (HC) were selected for the quantitative determination of their plasma metabolites using an ultraperformance liquid chromatography‐tandem mass spectrometry (UPLC‐MS/MS) platform. A total of 1997 metabolites were detected in the study cohort, from which 116 metabolites were found to be differentially expressed between the ASD and HC groups. Through analytical algorithms such as least absolute shrinkage selection operator (LASSO), support vector machine (SVM), and random forest (RF), three potential metabolic markers were identified as FAHFA (18:1(9Z)/9‐O‐18:0), DL‐2‐hydroxystearic acid, and 7(S),17(S)‐dihydroxy‐8(E),10(Z),13(Z),15(E),19(Z)‐docosapentaenoic acid. These metabolites demonstrated superior performance in distinguishing the ASD group from the HC group, as indicated by the area under curves (AUCs) of 0.935, 0.897, and 0.963 for the three candidate biomarkers, respectively. The samples were divided into training and validation sets according to 7:3. Diagnostic models were constructed using logistic regression (LR), SVM, and RF. The constructed three‐biomarker diagnostic model also exhibited strong discriminatory efficacy. These findings contribute to advancing our understanding of the underlying mechanisms involved in the occurrence of ASD and provide a valuable reference for clinical diagnosis.

Abstract Newborn screening (NBS) for isovaleric acidemia (IVA) is performed by flow injection tandem mass spectrometry quantifying C5 carnitines (C5). Isovalerylcarnitine, however, is isomeric with pivaloylcarnitine which can be present in blood due to maternal use of pivaloylester‐containing antibiotics, available in Germany since late 2016. During a 36‐month period (January 19–December 21), all newborns screened in Hamburg with a C5 above cutoff (NeoGram®: 0.50 μmol/L or Neobase®2: 0.45 μmol/L) were included in the study. As a second‐tier test, a simple ultra performance liquid chromatography‐tandem mass spectrometry (UPLC‐MS/MS) method was developed to differentiate the C5 isomers pivaloyl‐, 2‐methylbutyryl‐, isovaleryl‐, and valerylcarnitine. Out of 156 772 newborns tested, one turned out to have genetically proven IVA while 99 were false positive (C5: 0.5–8.2 μmol/L) due to the presence of pivaloylcarnitine. These cases have increased year by year and show local clusters. Retrospective analysis of another 39 cases from 287 206 neonates tested at the NBS center in Heidelberg with C5 elevation (0.9–10.6 μmol/L) but clinical and biochemical exclusion of IVA yielded evidence of pivaloylcarnitine in all cases. Inclusion of a second‐tier test into NBS significantly reduces the high and increasing false‐positive rate of IVA screening. This avoids further diagnostic steps, prevents unnecessary stress and anxiety of parents in a remarkably high number of cases. If Hamburg data of 2021 are extrapolated to all of Germany, one can assume around 800 (1‰) false‐positive cases in comparison to an average of two classic IVA cases per year. Unless licensing of pivaloylester‐containing drugs for use during pregnancy is reconsidered, a second‐tier test for C5 determination is indispensable.

Acetylcorynoline is an alkaloid isolated from the tubers of Corydalis ambigua Cham. et Schltdl. It has anti-inflammatory properties with the potential to treat Parkinson's disease. However, the use of UPLC-MS/MS for identifying acetylcorynoline in mouse plasma has not yet been explored. The present study aimed to develop a fast and selective method for determining the amount of acetylcorynoline in mouse plasma using UPLC-MS/MS. Plasma samples (10 μL) were prepared using methanol-induced protein precipitation following the addition of aconitine as an internal standard. The chromatographic separation was accomplished using a UPLC HSS T3 column with acetonitrile and 0.1% formic acid as the mobile phase. The analytes were run for 4.0 min in total. The target fragment ions m / z 410.4 ⟶ 350.3 for acetylcorynoline and m / z 646.6 ⟶ 586.5 for internal standard were used for quantification using multiple reaction monitoring mode. The mouse blood was obtained at different time points after intravenous (5 mg/kg) and oral (20 mg/kg) administration of acetylcorynoline. The calibration plots for acetylcorynoline in mouse plasma showed a linear trend over the whole range of 1-2000 ng/mL. Both the intraday and interday precision relative standard deviations were less than 11%. The half-life in mice was found to be 2.6 ± 0.7 h and 2.7 ± 0.8 h following oral and intravenous administration, respectively. The bioavailability was determined to be 58.9%. The pharmacokinetics and bioavailability of acetylcorynoline in mice were effectively analyzed using this UPLC-MS/MS method, which had a runtime of 4 min per sample and required only 10 μL of plasma., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2025 Mengzhi Xu et al. International Journal of Analytical Chemistry published by John Wiley & Sons Ltd.)

Deoxycholic acid (DCA) injection is applied in treating moderate to severe submental bulge or facial fullness caused by excessive submental fat accumulation. Using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) technology, which was swiftly, precisely, and reliably confirmed, DCA was determined in human plasma with low quantification limits of 56 ng/mL. We selected six healthy individual blank human plasma with low concentrations of endogenous DCA and mixed them to prepare standard curve samples. The samples were purified by the protein precipitation technique and then separated using a BEH C18 column (2.1 × 50 mm, 1.7 μm). Using multiple reaction monitoring (MRM) and electrospray ionization (ESI) sources operating in negative mode, the mass was identified and measured. The precursor-to-product ion transitions were observed at m/z 391.2 -  → 345.2 - and m/z 395.2 -  → 349.2 - for DCA and DCA-d 4 (isotope internal standard), respectively. This method was thoroughly validated, encompassing assessments of linearity, sensitivity, precision, selectivity, stability, matrix effect, accuracy, carryover, and recovery. In a word, the validation results demonstrated that this method exhibited sensitivity, accuracy, and reproducibility and could effectively be utilized for studying the pharmacokinetic properties of DCA in a randomized, parallel, controlled, Phase I clinical trial., (© 2025 John Wiley & Sons Ltd.)

Introduction: Vitex L. is a large genus of tropical and subtropical trees used in medicine of many nations. Some species are used in gynecology due to flavonoids, iridoids, and diterpenes. Other species were shown to contain ecdysteroids and are used as tonic remedies., Objective: This study aimed to investigate content and patterns of distribution of ecdysteroids in native trees and their accumulation in in vitro cell cultures as potential biotechnological agents of Vitex species growing in remote flora of Russia and Vietnam., Methodology: Targeted UPLC-MS/MS was used for the quantitation of the ecdysteroids in samples of plants and callus cultures of Vitex species. Determination of nucleotide sequences was carried out on ABI PRISM 3730xl sequencer. Phylogenetic tree was constructed using Mega v.7.0., Results: Samples of Vitex agnus-castus L. from Russian Black Sea coast of the Caucasus, as well as samples of nine species of Vitex from Vietnam were collected for ecdysteroids quantification. Relationships between the content of ecdysteroids and the systematic position of the Vitex species were shown., Conclusions: Higher levels of ecdysteroids were revealed in different plant parts of Vitex canescens Kurz, Vitex pinnata L., Vitex quinata (Lour.) F.N.Williams, and Vitex tripinnata (Lour.) Merr. It has been established that species rich in ecdysteroids, as well as those containing trace amount of ecdysteroids (V. agnus-castus L., Vitex negundo L., and Vitex rotundifolia L.f.) are grouped in certain clades isolated from each other on the molecular phylogenetic tree of the genus. The most promising raw materials are the young leaves of V. tripinnata (8562 μg/g of dry biomass), as well as the bark of V. canescens (16,520 μg/g) and V. quinata (19,130 μg/g). The bark of V. canescens could be a good source of turkesterone (11α-oxyecdysteroid) that has pronounced pharmacological effects. Its content reaches up to 1400 μg/g in the bark. Discovering ability of callus cultures for synthesis of ecdysteroids allows us to outline experimental ways to increase the level of ecdysteroid biosynthesis in biotechnological systems as alternative source of ecdysteroids., (© 2025 John Wiley & Sons Ltd.)

TNG908 is a potent and selective protein arginase methyltransferase 5 (PRMT5) inhibitor that is currently going through phase I/II clinical development for the treatment of non-small cell lung cancer. To facilitate pharmacokinetic and toxicokinetic studies of TNG908, here, we reported an ultra-high-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the detection of TNG908 in dogs. The dog plasma samples were precipitated by acetonitrile and analyzed using a Waters ACQUITY BEH C 18 column combined with a Thermo triple quadrupole mass spectrometer. The mobile phase consisted of 0.1% formic acid solution and acetonitrile, at a flow rate of 0.3 mL/min. TNG908 and internal standard were monitored by selective reaction monitoring (SRM) with m/z 410.2 > 150.1 and m/z 394.2 > 278.1, respectively. The method demonstrated excellent linearity over the concentration range of 1-1000 ng/mL, with a correlation coefficient greater than 0.995. Acetonitrile-mediated protein precipitation showed high extraction efficiency and a recovery above 80%. The validated assay was further applied to measure TNG908 in dog plasma after oral and intravenous administration and achieved success. The obtained pharmacokinetic parameters indicated low clearance of TNG908 (3.7 ± 0.8 mL/min/kg) and moderate oral bioavailability (>36.4%)., (© 2024 John Wiley & Sons Ltd.)

Introduction: Frankincense is used for analgesic, tumor-suppressive, and anti-inflammatory treatments in Traditional Chinese Medicine but poses toxicological concerns. Vinegar processing is a common technique used to reduce the toxicity of frankincense., Objective: This study aimed to investigate the chemical composition and quality evaluation of raw and vinegar-processing frankincense by multiple UPLC-MS/MS techniques. Additionally, we purposed refining the vinegar processing technique and identifying potentially harmful ingredients in the raw frankincense., Methodology: Sub-chronic oral toxicity studies were conducted on raw and vinegar-processing frankincense in rats. The composition of frankincense was identified by UPLC-Q-TOF-MS/MS. Chemometrics were used to differentiate between raw and vinegar-processing frankincense. Potential chemical markers were identified by selecting differential components, which were further exactly determined by UPLC-QQQ-MS/MS. Moreover, the viability of the HepG2 cells of those components with reduced contents after vinegar processing was assessed., Results: The toxicity of raw frankincense is attenuated by vinegar processing, among which vinegar-processing frankincense (R40) (herb weight: rice vinegar weight = 40:1) exhibited the lowest toxicity. A total of 83 components were identified from frankincense, including 40 triterpenoids, 37 diterpenoids, and 6 other types. The contents of six components decreased after vinegar-processing, with the lowest levels in R40. Three components, specifically 3α-acetoxy-11-keto-β-boswellic acid (AKBA), 3α-acetoxy-α-boswellic acid (α-ABA), and 3α-acetoxy-β-boswellic acid (β-ABA), inhibited the viability of HepG2 cells. The processing of frankincense with vinegar at a ratio of 40:1 could be an effective method of reducing the toxicity in raw frankincense., Conclusion: Our research improves understanding of the toxic substance basis and facilitates future assessments of frankincense quality., (© 2024 John Wiley & Sons Ltd.)

Ethoxysanguinarine (ETSG), a benzophenanthridine alkaloid, exhibits diverse biological activities, including antibacterial, antifungal, anti-inflammatory, antioxidant, and anti-tumor effects. Despite these properties, limited research exists on ETSG in vivo pharmacokinetics due to its poor solubility and low bioavailability. In this study, we developed a rapid and specific UPLC-MS/MS method for ETSG bioanalysis. Sample preparation involved one-step protein precipitation using methanol and phellodendrine as an internal standard (IS). The Waters HSS T3 column (2.1 * 50 mm, 1.8 μM) employed a gradient elution with mobile phases A (2 mmol/L ammonium formate aqueous solution-formic acid [99.8:0.2, v/v]) and B (methanol-formic acid [99.8:0.2, v/v]). Mass analysis via Waters Q-mass spectrometer utilized positive scan mode and multiple reaction monitoring. ETSG and IS were detected at m/z 332.0 → 274.0 and 342.0 → 177.0, respectively, within 7.0 min. The method demonstrated excellent precision, accuracy, recovery, and stability, with a linear calibration curve (1.1-560 ng/mL) and strong correlation coefficient (0.9984). Successful pharmacokinetic evaluation in Sprague-Dawley rats included intravenous ETSG administration and intragastric ETSG nanoemulsion/suspension. This method enables steroidal saponin analysis from ETSG in biological samples., (© 2025 John Wiley & Sons Ltd.)

Abstract Niazirin is a nitrile glycoside isolated from the pods and leaves of Moringa oleifera and possesses antidiabetic activity. However, its pharmacokinetic properties have not been well studied yet. The present study aims to establish a specific, rapid, and simple method for the determination of niazirin in rat plasma and apply it to evaluate its absolute bioavailability and dose proportionality. MS/MS detection was performed in multiple reaction monitoring modes with negative electrospray ionization. The precursor–product ion transitions were m/z 278.1 [M‐H]− → 130.8 for niazirin and m/z 316.9 [M‐H]− → 271.0 for IS. The intra‐ and inter‐day precision (RSD) was

Pectolinarin is a flavonoid compound known for its wound-healing properties, including anti-inflammatory and antibacterial effects. In this study, we employed UPLC-MS/MS to quantify pectolinarin in rat plasma and investigate its pharmacokinetics. Plasma samples were processed using an acetonitrile precipitation method. Chromatographic separation was performed on a UPLC BEH column with a gradient mobile phase of acetonitrile-water (containing 0.1% formic acid). Detection was carried out using electrospray ionization (ESI) tandem mass spectrometry in multiple reaction monitoring (MRM) mode with positive ionization, targeting transitions of m/z 623.3 → 315.3 for pectolinarin and m/z 370.5 → 125.0 for the IS. The results demonstrated that pectolinarin exhibited acceptable linearity in rat plasma within the concentration range of 1.2 to 2300 ng/mL (r > 0.995). The intraday and interday precision, expressed as relative standard deviation (RSD), was below 9.2%. Accuracy ranged from 97.3% to 108.3%, with average recovery exceeding 94.7%. The matrix effect was between 97.8% and 105.3%. The method was successfully applied to evaluate the pharmacokinetics of pectolinarin in rats following both oral and intravenous administration. The absolute bioavailability of pectolinarin in rats was determined to be 0.28%., (© 2025 John Wiley & Sons Ltd.)

A sensitive and simple method using ultra-liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed and validated to determine the concentration of curcumin in rat plasma and tissue samples. Emodin was selected as the internal standard (IS), and biological samples were pretreated with simple one-step acetonitrile precipitation. The calibration curves exhibited linearity within the range of 1-1000 ng/ml for both rat plasma and tissue samples. The accuracy and precision of intra-day as well as inter-day determinations ranged from 99.3% to 117.3% and from 98.2% to 105.1%, respectively. This method demonstrated excellent recovery rates ranging from 76.4% to 96.4% along with minimal matrix effect ranging from 86.5% to 99.6%. The effectiveness of this method was successfully demonstrated through its application in an in vivo pharmacokinetic and tissue distribution study after single administration via inhalation (100 mg/kg), oral gavage (100 mg/kg) and intravenous injection (2.5 mg/kg) of curcumin in rats. The results revealed that inhalation significantly improved the bioavailability of curcumin, with most of the drug being deposited in the lung. These findings highlight inhalation as an effective route for targeted delivery of drugs directly into lung tissues, thus suggesting potential future applications for treating pulmonary diseases utilizing inhaled curcumin., (© 2024 John Wiley & Sons Ltd.)

Yigong San (YGS) is a traditional Chinese medicine formula used for pediatric anorexia, chronic atrophic gastritis, and irritable bowel syndrome. In this study, the excretion of eight main compounds, including liquiritin; isoliquiritin; hesperidin; ginsenosides Rb 1 , Re, and Rg 1 ; and atractylenolides I and II, in rat urine, feces, and bile, was investigated by ultra-high performance liquid chromatography-tandem mass spectrometry. The results showed that the cumulative excretion rates of the compounds in rat urine, feces, and bile were 0.018-1.15%, 0.024-19.89%, and 0.0025-0.72%, respectively. Among the eight compounds detected, liquiritin was the richest in urine, and ginsenosides Re and Rg 1 and atractylenolide I were mainly found in feces and bile. In summary, the main components of YGS are excreted via multiple approaches. Liquiritin is mainly through urine, whereas isoliquiritin; hesperidin; ginsenosides Rb 1 , Re, and Rg 1 ; and atractylenolides I and II are mainly through feces. The excretion of these compounds in bile is usually positively correlated with that in feces. This study lays a foundation for further pharmacological research and application of YGS., (© 2024 John Wiley & Sons Ltd.)

Huperzine A is a reversible and selective cholinesterase inhibitor and has been approved for the treatment of Alzheimer's diseases. In this study, we developed a highly sensitive and specific ulta-high-performance liquid chromatography-tandem mass spectrometry method for the determination of Huperzine A in rat plasma. An aliquot of 50 μL of rat plasma sample was pretreated with 200 μL of acetonitrile-methanol (v/v; 1:1) containing 0.2% formic acid followed by solid phase extraction. The resulting sample was separated on a Waters ACQUITY BEH C 18 column using acetonitrile and water containing 0.2% formic acid as mobile phase, at a flow rate of 0.3 mL/min. Multiple-reaction monitoring (MRM) mode was used for quantitative analysis of Huperzine A in positive electrospray ionization. In the concentration range of 0.01-10 ng/mL, Huperzine A showed excellent linearity with correlation coefficient > 0.998. The intra- and inter-day RSD% were less than 9.7%, while the RE% ranged from -6.7% to 10.0%. The mean recovery was >84.5%. The validated method was demonstrated to be selective, sensitive, and reliable, which has been successfully applied to pharmacokinetic study of Huperzine A in rat plasma. Huperzine A displayed a long half-life in rat plasma and high oral bioavailability., (© 2024 John Wiley & Sons Ltd.)

Dissipative behavior and final residue levels of difenoconazole, prochloraz, propiconazole, and pyraclostrobin in figs were investigated using field trials and laboratory assays. A three-factor, three-level orthogonal test was designed to optimize the pretreatment conditions of the method. A method was established using high-performance liquid chromatography tandem mass spectrometry for the determination of difenoconazole, prochloraz, propiconazole, and pyraclostrobin residues in figs. The limit of quantification for all four targets in figs was 0.002 mg/kg. Difenoconazole, prochloraz, propiconazole, and pyraclostrobin are readily digestible pesticides in figs with half-lives of 6.4, 6.2, 4.8, and 7.9 days, respectively. Residues of difenoconazole, prochloraz, propiconazole, and pyraclostrobin in figs were below the European Union established residue levels of 0.1, 0.03, 0.01, and 0.02 mg/kg, respectively, at day 7 after application. Pyraclostrobin, propiconazole, difenoconazole, and prochloraz were applied twice at doses of 75, 125, 150, and 200 mg a.i./kg at 7-day intervals, and the residues of the four fungicides in figs were acceptable 7 days after the last application. Therefore, the safety interval can be set at 7 days for 70% difenoconazole-prochloraz wettable powder and 40% pyraclostrobin-propiconazole aqueous emulsion according to the protocol., (© 2024 John Wiley & Sons Ltd.)

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by the disruption of synaptic communication among millions of neurons. Recent research has highlighted the potential therapeutic effectiveness of natural polyphenolic compounds in addressing AD. Soybeans are abundant in polyphenols, and their polyphenolic composition undergoes significant alteration through fermentation by Eurotium cristatum. Through comprehensive database searches, we identified active components within fermented soybean polyphenols and genes associated with AD. Subsequently, we utilized Venn diagrams to analyze the overlap between AD-related genes and these components. Furthermore, we visualized the network between intersecting targets and proteins using Cytoscape software. The anti-AD effects of soybeans were further explored through comprehensive analysis, including protein-protein interaction analysis, pathway enrichment analysis, and molecular docking studies. Our investigation unveiled 6-hydroxydaidzein as a major component of fermented soybean polyphenols, shedding light on its potential therapeutic significance in combating AD. The intersection between target proteins of fermented soybeans and disease-related targets in AD comprised 34 genes. Protein-protein interaction analysis highlighted key potential targets, including glyceraldehyde-3-phosphate dehydrogenase (GAPDH), glycogen synthase kinase 3 beta (GSK3B), amyloid precursor protein (APP), cyclin-dependent kinase 5 (CDK5), and beta-site APP cleaving enzyme 1 (BACE1). Molecular docking results demonstrated a robust binding effect between major components from fermented soybeans and the aforesaid key targets implicated in AD treatment. These findings suggest that fermented soybeans demonstrate a degree of efficacy and present promising prospects in the prevention of AD., (© 2024 John Wiley & Sons Ltd.)

The present study examined the pharmacokinetics of IMM-H012 in rat plasma, utilizing ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Internal standard cilostazol was employed, and plasma samples were processed using acetonitrile precipitation. A mobile phase (acetonitrile-0.1% formic acid in water) with gradient elution was used to achieve chromatographic separation using a UPLC BEH C18 column. In multiple reaction monitoring mode, electrospray ionization MS/MS was utilized in positive ionization mode. Based on findings, the lower limit of quantification was 2 ng/mL, and the linearity of IMM-H012 in rat plasma was found to be acceptable within the range of 2-2000 ng/mL (R 2  > 0.995). The intra-day and inter-day precision relative standard deviation was less than 14% of IMM-H012 in rat plasma. The matrix effect was within the range of 102%-107%, and the accuracy ranged from 92% to 113%. Pharmacokinetics of IMM-H012 in rats after oral administration were successfully studied using UPLC-MS/MS., (© 2024 John Wiley & Sons Ltd.)

VX-548 is an orally active and highly selective NaV 1.8 inhibitor that is undergoing development for the treatment of acute pain. The aim of this study was to develop a liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the measurement of VX-548 in monkey plasma. VX-548 was extracted from the plasma using acetonitrile-mediated protein precipitation. The quantitative analysis was performed on a Thermo Vantage TSQ mass spectrometer with ibrutinib as an internal standard. Chromatography was performed on a Waters ACQUITY UPLC BEH C 18 column with 0.1% aqueous formic acid and acetonitrile as mobile phase. The precursor-to-product ion transitions were m/z 474.2 > 165.0 and m/z 441.2 > 138.1 for VX-548 and internal standard, respectively. This developed method was successfully validated in the concentration range of 1-1000 ng/mL. The calibration curve showed excellent linearity with a correlation coefficient of >0.999. The precision expressed as relative standard deviation (RSD) was <8.4%, whereas the accuracy denoted as relative error (RE) ranged from -5.0% to 9.1%. The mean recovery was >84%. VX-548 was stable in monkey plasma after storage under certain conditions. The validated method was successfully applied to the pharmacokinetic study of VX-548 in monkey plasma after single oral (2 mg/kg) and intravenous (1 mg/kg) administrations., (© 2024 John Wiley & Sons Ltd.)

Numerous studies have suggested that intra-articular administration of antibiotics following primary revision surgery may be one of the methods for treating prosthetic joint infection (PJI). Vancomycin and meropenem are the two most commonly used antibiotics for local application. Determining the concentrations of vancomycin and meropenem in the serum and synovial fluid of patients with PJI plays a significant role in further optimizing local medication schemes and effectively eradicating biofilm infections. This study aimed to establish a rapid, sensitive, and accurate ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for determining the concentrations of vancomycin and meropenem in human serum and synovial fluid. Serum samples were processed using acetonitrile precipitation of proteins and dichloromethane extraction, while synovial fluid samples were diluted before analysis. Chromatographic separation was achieved in 6 min on a Waters Acquity UPLC BEH C18 column, with the mobile phase consisting of 0.1% formic acid in water (solvent A) and acetonitrile (solvent B). Quantification was carried out using a Waters XEVO TQD triple quadrupole mass spectrometer with an electrospray ionization (ESI) source in positive ion mode. The multiple reaction monitoring (MRM) mode was employed to detect the following quantifier ion transitions: 717.95-99.97 (norvancomycin), 725.90-100.04 (vancomycin), 384.16-67.99 (meropenem). The method validation conformed to the guidelines of the FDA and the Chinese Pharmacopoeia. The method demonstrated good linearity within the range of 0.5-50 μg/ml for serum and 0.5-100 μg/ml for synovial fluid. Selectivity, intra-day and inter-day precision and accuracy, extraction recovery, matrix effect, and stability validation results all met the required standards. This method has been successfully applied in the pharmacokinetic/pharmacodynamic (PK/PD) studies of patients with PJI., (© 2024 John Wiley & Sons Ltd.)

In this paper, an ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for quantifying the levels of crassicauline A, fuziline, karacoline, and songorine in rat plasma. After processing the rat plasma, the proteins in the plasma were separated by extracting the analytes with acetonitrile-methanol (9:1, v/v). The chromatographic column used was the UPLC HSS T3 column, and the mobile phase (methanol-water with 0.1% formic acid) under a gradient elution profile was used to separate the four compounds, with elution times for each analyte being less than 5 min. Electrospray ionization in positive-ion mode and operating in multiple reaction monitoring mode was used for quantitative analysis. Crassicauline A, fuziline, karacoline, and songorine were administered to 48 rats (n = 6 per group) orally (5 mg/kg) and intravenously (0.5 mg/kg). The standard curves demonstrated excellent linearity in the range of 1-2500 ng/mL, wherein all r values were greater than 0.99. The UPLC-MS/MS method for the determination of crassicauline A, fuziline, karacoline, and songorine in rat plasma was successfully applied in determining their pharmacokinetics parameters, from which their oral bioavailabilities were calculated to be 18.7%, 4.3%, 6.0%, and 8.4%, respectively., (© 2024 John Wiley & Sons Ltd.)

Introduction: Ginkgo biloba extract (GBE) is an effective substance from traditional Chinese medicine (TCM) G. biloba for treating ischaemic stroke (IS). However, its active ingredients and mechanism of action remain unclear., Objectives: This study aimed to reveal the potential active component group and possible anti-IS mechanism of GBE., Materials and Methods: The network pharmacology method was used to reveal the possible anti-IS mechanism of these active ingredients in GBE. An ultra-high-performance liquid chromatography triple quadrupole electrospray tandem mass spectrometry (UPLC-MS/MS) method was established for the simultaneous detection of the active ingredients of GBE., Results: The active components of GBE anti-IS were screened by literature integration. Network pharmacology results showed that the anti-IS effect of GBE is achieved through key active components such as protocatechuic acid, bilobalide, ginkgolide A, and so on. Gene Ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that the possible anti-IS mechanism of GBE is regulating the PI3K-Akt signalling pathway and other signal pathways closely related to inflammatory response and apoptosis regulation combined with AKT1, MAPK, TNF, ALB, CASP3, and other protein targets. Nineteen main constituents in seven batches of GBE were successfully analysed using the established UPLC-MS/MS method, and the results showed that the content of protocatechuic acid, gallic acid, ginkgolide A, and so forth was relatively high, which was consistent with network pharmacology results, indicating that these ingredients may be the key active anti-IS ingredients of GBE., Conclusion: This study revealed the key active components and the anti-IS mechanism of GBE. It also provided a simple and sensitive method for the quality control of related preparations., (© 2023 John Wiley & Sons, Ltd.)

In this present study, we developed a reliable and simple ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) assay for the simultaneous quantification of paeoniflorin, albiflorin, oxypaeoniflorin, benzoylpaeoniflorin and isomaltopaeoniflorin in beagle dog plasma. We also analyzed the pharmacokinetics of those components after oral administration of fried Radix Paeoniae Alba (FRPA) in beagle dogs. Plasma samples were processed by protein precipitation with methanol. Chromatographic separation was performed with a Waters HSS-T3 C 18 column (100 × 2.1 mm, 1.8 μm, kept at 40°C) using multiple reaction monitoring mode. A gradient elution procedure was used with solvent A (0.02% formic acid-water) and solvent B (0.02% formic acid-acetonitrile) as mobile phases. Method validation was performed as US Food and Drug Administration guidelines, and the results met the acceptance criteria. The method we establish in this experiment was successfully applied to the pharmacokinetic study after oral administration of FRPA extract to beagle dogs., (© 2023 John Wiley & Sons, Ltd.)

Although levetiracetam (LEV) has favorable linear pharmacokinetic properties, therapeutic drug monitoring (TDM) is necessary for pregnant women with epilepsy. This study aims to build a simple, reliable, and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for determining LEV concentrations in plasma and saliva samples, to support the routine TDM of LEV in Chinese pregnant women with epilepsy. The stable isotope-labeled LEV-d 6 was used as the internal standard. The extracted samples were analyzed using a UPLC-MS/MS system with positive electrospray ionization. Mobile phase A was water containing 5 mM ammonium acetate and 0.1% formic acid, and phase B was 1:1 methanol-acetonitrile with 0.1% formic acid. The method was validated and utilized to determine LEV concentrations in non-pregnant and pregnant patients with epilepsy. The developed method was validated in both plasma and saliva samples over a concentration range of 0.1-50 μg/mL. The intra- and inter-batch accuracy for LEV ranged from -7.0% to 2.9%, with precisions between 2.7% and 9.3%. In pregnant patients, the mean dose-standardized LEV trough plasma concentrations were significantly lower than those in non-pregnant patients (4.73 ± 2.99 vs. 7.74 ± 3.59 ng/mL per mg/day; P < 0.0001). It is recommended that the TDM of LEV should be routinely performed during the different stages of pregnancy., (© 2023 John Wiley & Sons, Ltd.)

This study performed the simultaneous quantification of assay and two alkyl sulfonate (tosylate) analogs of empagliflozin (EGZ), specifically methyl 4-methyl benzene sulfonate (MMBS) and ethyl 4-methyl benzene sulfonate (EMBS) in EGZ, and its finished dosage form using an accurate and sensitive ultra-performance liquid chromatography-mass spectrometry method. The separation was achieved on a Waters Acquity BEH Shield RP18 (100 × 2.1 mm, 1.7 μm) column in gradient elution mode with 0.1% formic acid and acetonitrile as the mobile phases and a flow rate of 0.5 mL/min. For simultaneous quantification, the multiple reaction monitoring technique was utilized. The procedure was successfully validated in accordance with the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines. The peak areas of both impurities, along with their concentrations, exhibited a good relationship with Pearson's correlation coefficient (R), which was >0.999 in the range of 0.3-6 ppm with an EGZ concentration of 2 mg/mL. The percentage recoveries from the limit of quantitation (LOQ) to 200% to the specification level were in the range of 94.82%-102.92%, whereas the percentage relative standard deviation (%RSD) was <2. Therefore, this method is rapid and accurate to quantify MMBS, EMBS, and EGZ assay simultaneously from the marketed tablet dosage forms of EGZ for commercial release and stability sample testing., (© 2023 John Wiley & Sons, Ltd.)

In this study, an ultra-performance liquid chromatography-tandem mass spectrometry method was established for the development and validation of baloxavir acid (BXA) concentrations and the active ingredients of the antiviral drug baloxavir marboxil (BXM). Further, the method was applied to study the pharmacokinetics of BXA. BXA was determined by the electrospray ionization multiple reaction monitoring positive ion mode, and the mass-to-charge ratios (m/z) of BXA and internal standard baloxavir-d4 were 484.2 → 247.2 and 488.1 → 247.2. An Oasis max online column (2.1 × 20 mm, 30 μm) was used with 1% formic acid in water (A) and 2% formic acid in acetonitrile (B) as mobile phases at a flow rate of 0.5 mL·min -1 for chromatographic separation. The linearity was good in the range of 3-200 ng·mL -1 (r = 0.9994), with 3.00 ng·mL -1 lower limit of quantification. The relative standard deviation of the inter-assay precision was less than or equal to 6.51%, and the accuracy was in the range of 91.28%-104.29%. This method is suitable for the determination of BXA and for performing pharmacokinetic studies in clinical research., (© 2023 John Wiley & Sons, Ltd.)

Favipiravir, a broad-spectrum RNA-dependent RNA polymerase inhibitor, is currently being evaluated in preclinical and clinical studies for the treatment of various infectious diseases including COVID-19. We developed an ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) assay for the quantification of favipiravir and its hydroxide metabolite (M1), in human and hamster biological matrices. Analytes were separated on an Acquity UPLC HSS T3 column (2.1 × 100 mm, 1.8 μm) after a simple protein precipitation with acetonitrile. The mobile phase consisted of water and methanol, each containing 0.05% formic acid. Experiments were performed using electrospray ionization in the positive and negative ion mode, with protonated molecules used as the precursor ion and a total run time of 6 min. The MS/MS response was linear over the concentration ranges from 0.5-100 μg/ml for favipiravir and 0.25-30 μg/ml for M1. Intra- and inter-day accuracy and precision were within the recommended limits of the European Medicines Agency guidelines. No significant matrix effect was observed, and the method was successfully applied to inform favipiravir dose adjustments in six immunocompromised children with severe RNA viral infections. In conclusion, the UPLC-MS/MS assay is suitable for quantification of favipiravir over a wide range of dosing regimens, and can easily be adapted to other matrices and species., (© 2023 The Authors. Biomedical Chromatography published by John Wiley & Sons Ltd.)

The hexosamine biosynthesis pathway (HBP) is a glucose metabolism pathway that produces uridine diphosphate N-acetyl glucosamine (UDP-GlcNAc). Substantial changes in HBP, including elevated HBP flux and UDP-GlcNAc levels, are associated with cancer pathogenesis. Particularly, cancer cells expressing oncogenic Kirsten rat sarcoma virus (KRAS) are highly dependent on HBP for growth and survival. To differentiate between HBP metabolites in KRAS wild-type (WT) and mutant (MT) lung cancer cells, a simultaneous quantitative method for analyzing seven HPB metabolites was developed using ultra-high-performance liquid chromatography-tandem mass spectrometry. A simple method without complicated preparation steps, such as derivatization or isotope labeling, was optimized for the simultaneous analysis of highly hydrophilic HBP metabolites, and the developed method was successfully verified. The intra- and inter-day coefficients of variation were less than 15% for all HBP metabolites, and the recovery was 89.67-114.5%. All results of the validation list were in accordance with ICM M10 guidelines. Through this method, HBP metabolites in lung cancer cells were accurately quantified, and it was confirmed that all HBP metabolites were upregulated in KRAS MT cells compared with KRAS WT lung cancer cells. We expect that this will be a useful tool for metabolic research on cancer and for the development of new drugs for cancer treatment., (© 2023 John Wiley & Sons Ltd.)

A simple and sensitive method using in vivo microdialysis coupled with UPLC-MS/MS was established to evaluate the pharmacokinetics of Shuang Hu tincture (SHZTN). Xevo TQ-S was used to analyze the active ingredients of mesaconitine, hypaconitine, 4-hydroxycinnamic acid, ferulic acid and N-(2, 3-dimethyl phenyl)-2- aminobenzoic acid of SHZTN. Samples were prepared using a methanol precipitation method and the internal standards lannaconitine and p-hydroxybenzoic acid were added. The method validation was conducted according to the guidelines of the Pharmacopoeia of China. A good linear range was obtained in the range of 1-2,000 ng/ml. The intra-day and inter-day precisions were less than 14.7%, and the accuracy range of all the analytes was -10.5-9.3%. The recovery of each analyte was over 95.5%, and matrix effects can be neglected. After a single dose of 20 mg/kg SHZTN, the area under the curve and peak concentration of the five active ingredients were significantly increased by transdermal compared with oral administration, which indicated the high bioavailability of SHZTN. The time to peak concentration of all compounds was <3.4 h, and the half-life was <15.4 h, which indicated that the five compounds have the best absorption and rapid elimination. The method was successfully developed and applied to the pharmacokinetic study of SHZTN., (© 2022 John Wiley & Sons Ltd.)

A simple, efficient, and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for simultaneous determination and pharmacokinetic study of salvianolic acid D, rosmarinic acid, lithospermicic acid, salvianolic acid B, and salvianolic acid Y in rat plasma after intravenous administration of salvianolic acid for injection. Three doses of administration, containing 10, 25, and 62.5 mg/kg, were investigated. Plasma samples were pretreated using protein precipitation with pre-cooled acetonitrile. As shown in S1, Chromatographic separation was achieved on a Waters Acquity UPLC® BEH C18 column (1.7 μm, 2.1 × 100 mm) with a mobile phase composed of acetonitrile-methanol-0.5% aqueous formic acid (10:30:60, v/v/v) at a flow rate of 0.3 ml/min. MS was detected by electrospray ion source negative ion mode and multiple reaction monitoring mode. The method was fully validated. The calibration curves for the five phenolic acids were linear in the given concentration ranges. The extraction recoveries, matrix effects, intra-day and inter-day precisions, and accuracies of the five analytes were all within acceptable limits. No significant difference of elimination half-life time (T 1/2 ) of five analytes at three doses was observed. Area under the curve and peak concentration (C max ) of the five analytes demonstrated a linear increase in the doses with the linear correlation r of each analyte at three doses being greater than 0.915. It indicated that the pharmacokinetic behavior of is positively related to the dose in the range of 10-62.5 mg/kg., (© 2022 John Wiley & Sons Ltd.)

This study aims to assess the bioequivalence of test and reference formulations of trimetazidine dihydrochloride in healthy Chinese volunteers under fasting and fed conditions, and to determine the effect of food on the pharmacokinetic profiles of both formulations. A randomized, open-label, crossover, four-period study with a 7-day washout period was conducted in 24 healthy Chinese subjects. The subjects fasted for at least 10 hours before being given a single 35-mg dose of the test and reference tablets. Venous blood samples were taken from predose at 0 hours to postdose at 36 hours at scheduled time points. The main pharmacokinetic parameters were calculated with a noncompartmental model. The nonparametric test of T max under both conditions showed no significant difference between the two formulations (P > .05). The 90% confidence intervals of geometric mean ratio of lnC max and lnAUC 0→∞ (the logarithmic values of area under the plasma concentration-time curve [AUC] and mean maximum plasma concentration [C max ]) all fell within 80%-125%. C max in the fed state was slightly higher than that in the fasting state (P < .05), while other pharmacokinetic parameters were comparable. No severe adverse events occurred. The test and reference formulations were bioequivalent under both fasting and fed conditions. Food did not affect the pharmacokinetic profiles of trimetazidine in Chinese healthy volunteers, therefore trimetazidine is suitable for administration under fasting or fed conditions., (© 2022, The American College of Clinical Pharmacology.)

Caged polyprenylated xanthones are the main active ingredients isolated from the resin of Garcinia hanburyi, which has been reported to exhibit potential anticancer and anti-inflammatory activities. This study aimed to develop sensitive and specific ultra-performance liquid chromatography coupled with the triple quadrupole mass spectrometry method for investigating the tissue distribution of five xanthones in rats: β-morellic acid, isogambogenic acid, gambogenic acid, R-gambogic acid and S-gambogic acid. All tissue samples were prepared using the liquid-liquid extraction method and separated on a C 8 column with a gradient system. Detection was performed on a triple quadrupole mass spectrometer in multiple-reaction monitoring using positive ionization. The method established in this assay was successfully applied to the tissue distribution study of the five selected xanthones after orally administering crude and processed gamboge in rat tissues. The results indicated that these five xanthones were distributed to rat tissues rapidly and could be detected in all of the selected tissues after oral administration. After processing, the contents of R-gambogic acid and S-gambogic acid in the gastrointestinal tract were significantly reduced. The findings of this study might be helpful in further understanding the processing mechanism of gamboge and providing references for its reasonable clinical application., (© 2022 John Wiley & Sons Ltd.)

Almonertinib was included in the first-line treatment of non-small cell lung cancer with EGFR T790M mutations by the Chinese Society of Clinical Oncology in 2021. Considering that immunocompromised lung cancer patients are prone to opportunistic fungal infections, and most triazole antifungal drugs are moderate or strong inhibitors of CYP3A4, this study was conducted to develop and validate an accurate and rapid ultra-performance liquid chromatography tandem mass spectrometry method for quantifying almonertinib in plasma and for investigating the pharmacokinetic changes of almonertinib caused by voriconazole and fluconazole in rats. After liquid-liquid extraction with tert-butyl methyl ether, an XSelect HSS T3 column (2.1 × 100 mm, 2.5 μm, Waters) was used for the chromatographic separation of almonertinib and sorafenib-D 3 (internal standard). The analytes were detected using an AB Sciex Triple Quad 5,500 mass spectrometer in the positive ionization mode. The method exhibited great linearity (0.5-200 ng/ml, r > 0.997) and stability under the established experimental conditions. All validation experiments were in accordance with the guidelines, and the results were all within the acceptable limits. This method was successfully applied to the researches of pharmacokinetics and drug interactions for almonertinib in rats. Voriconazole and fluconazole significantly altered the pharmacokinetic profiles of almonertinib and increased the systemic exposure of almonertinib in rats to different degrees, but further human trials should be conducted to validate the results., (© 2022 John Wiley & Sons Ltd.)

Recent studies have revealed that the combination therapy of atorvastatin (ATV) with naringenin (NG) can offer meaningful benefits in the treatment of hypercholesterolemia, while decreasing adverse side effects. To investigate whether there are pharmacokinetic interactions among ATV, its metabolite 2-hydroxy atorvastatin (2-ATV), and NG, in the current study, we developed and validated a simple, rapid, and specific UPLC-MS/MS method to simultaneously determine the concentrations of these analytes in the rat plasma. Sample preparation was performed using simple protein precipitation. Chromatographic analysis was carried out on an Acquity UPLC BEH C18 column (1.7 μm, 2.1 × 100 mm) using gradient elution mode, and these three analytes were detected using a Xevo® TQD triple quadrupole tandem mass spectrometer, in the positive ion electrospray ionization interface. The developed method showed good linearity over the following concentrations in rat plasma samples: 3-1200 ng/ml (r = 0.9965) for ATV, 1.5-600 ng/ml (r = 0.9934) for 2-ATV, and 3-1200 ng/ml (r = 0.9964) for NG. The assays were validated and satisfied the acceptance criteria recommended by U.S. Food and Drug Administration guidelines. Upon successful application of the method to a pharmacokinetic interaction study, the results indicated that NG significantly enhanced the bioavailability of ATV and 2-ATV., (© 2022 John Wiley & Sons Ltd.)

A reliable and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the determination of zanubrutinib in the plasma of beagle dogs. The column used was an Acquity BEH C 18 column (2.1 mm × 50 mm, 1.7 μm), maintained at 40°C with an injection volume of 2 μl. The gradient elution program was as follows: 0-1 min, 10-10% A; 1-1.1 min, 10-90% A; 1.1-2.1 min, 90-90% A; 2.1-2.2 min, 90-10% A; 2.2-3.0 min, 10-10% A. Mobile phase A was 0.1% formic acid, B was acetonitrile, and the total analysis time was 3 min. The mass spectrometry was performed in positive ion mode, and the scanning mode was multi-reaction monitoring mode with electrospray ionization as the ion source; m/z 472.2 → 455.01 for zanubrutinib and m/z 441.03 → 137.99 for ibrutinib (internal standard). The plasma samples were processed by protein precipitation. The standard curve showed good linearity (r 2  = 0.999 8) in the range of 1.0-1,000 ng/ml (zanubrutinib) with a low limit of quantification of 1 ng/ml. Also, the intra-day and inter-day precision (RSD) was <5.88% and the accuracy (RE) ranged from -1.56 to 1.08%; the recoveries of zanubrutinib in beagle plasma ranged from 90.12 to 93.53% (RSD 1.67-6.42%) and the ME values of zanubrutinib were 98.70-101.06% (RSD 5.37-8.49%, n = 6). All values meet US Food and Drug Administration requirements. A rapid, highly selective and sensitive method for the determination of zanubrutinib concentration in plasma by UPLC-MS/MS was successfully developed. This method is suitable for pharmacokinetic studies in beagle dogs by following oral administration of zanubrutinib., (© 2022 John Wiley & Sons Ltd.)

Luhong recipe (LHR) is has been used as an empirical prescription for treating chronic heart failure for long, with safety, reliability, and significant efficacy. However, its pharmacokinetics has not yet been studied. This study aims to establish a ultra performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous analysis of epimedin A, epimedin B, epimedin C, icariin, psoralen, and isopsoralen in rat plasma and apply it to the pharmacokinetic study of LHR after oral administration. These six analytes were ionized using positive electrospray ionization (ESI + ). The MS/MS transitions used for monitoring are successively converted to m/z 839.3 → 369.1, m/z 809.2 → 369.1, m/z 823.3 → 369.1, m/z 677.2 → 205.2, m/z 187.1 → 115.2, and m/z 230 → 120.9. Linearity, precision, accuracy, stability, matrix effect, and recovery of the established method were within the acceptable range. The method was suitable for the determination of six analytes after oral administration of LHR. The pharmacokinetic results showed that the time to reach the peak concentration (T max ) was from 0.17 to 13.5 h, the peak concentration (C max ) was 109.23-980 ng/mL, the area under the concentration-time curve (AUC [0 - t] ) was 65.48-8846.08 ng·h/mL, and the apparent distribution volume (Vd) was 24,772-896,132 mL/kg. These results provided a meaningful basis for formulating the clinical dose regimen of LHR., (© 2022 John Wiley & Sons Ltd.)

Astragalus Membranaceus (AM) is widely applied in Chinese herbal compound formulas for treating various kinds of diseases. However, relative pharmacokinetics data on AM in nonrodents is still lacking. Here, an UPLC-MS/MS method for determining the six main compounds of AM was developed. The chromatographic separation was carried out by a Waters Acquity UPLC HSS T3 column (100 × 2.1 mm, 1.8 μm) with gradient elution of water-formic acid (99.98:0.02, v/v) and acetonitrile-formic acid (99.98:0.02, v/v) at a flow rate of 0.3 ml/min within 11 min. Analyses of all compounds were conducted in multiple reaction monitoring mode with a positive/negative ion-switching mode of an electrospray ionization source in a single run. The analytical method was validated in terms of specificity, linearity, accuracy, precision, stability, etc. The method showed excellent linearity (r > 0.999) over certain concentration ranges. The intra-day and inter-day precisions were evaluated, and the RSD values were <12.4%. Furthermore, the validated method was successfully applied to determine the six components in plasma after oral administration of AM aqueous extract to beagle dogs and the pharmacokinetic parameters were obtained. Together, this study provides a reference for medication in the clinical practice of AM., (© 2022 John Wiley & Sons Ltd.)

Mahuang Xuanfei Zhike (MXZ) syrup, a Chinese patent medicine, has been widely used in the clinical treatment of cough. However, there is no reported method for the quantitative analysis of the effective components of MXZ syrup in biological samples. In this study, the effective components of MXZ syrup were screened by network pharmacology and molecular docking technology. A sensitive and rapid ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established to test the active components of MXZ syrup in rat plasma and tissue homogenates, including ephedrine, amygdalin, chlorogenic acid, harpagoside, forsythin and forsythoside A. Chromatographic separation was performed on a Waters Acquity UPLC HSS T3 column (2.1 × 50 mm, 1.8 μm) and the mass analysis was conducted using a Waters Xevo TQ mass spectrometer using multiple reaction positive and negative ion simultaneous monitoring mode. The results showed that the linearity ranged from 0.3 to 409.4 ng/ml. The extraction recoveries were all <8.33%, and the matrix effects were all <8.45, which met the requirements. The pharmacokinetic and tissue distribution results indicated that the main active components of MXZ syrup were absorbed quickly and eliminated slowly in vivo, and there may be a reabsorption process., (© 2022 John Wiley & Sons Ltd.)

SHR0302, as a novel Janus kinase (JAK) inhibitor 1, is used for treatment of rheumatoid arthritis (RA) in humans. A novel and sensitive ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) has been developed and validated for determining the concentration of SHR0302 in human plasma. A precipitation deproteinization method was used for plasma pretreatment with methanol. Detection was carried out on an Agilent 1,260 UPLC coupled with a Triple Quad 4000 mass spectrometer operated in positive multiple reaction monitoring mode, and the analytes were separated on a Synergi Polar-RP C 18 (50 × 2.0 mm, 4 μm, Phenomenex) analytical column with gradient elution of 0.1% formic acid, and 2 mmol/l ammonium acetate in water and 0.1% formic acid and 2 mmol/l ammonium acetate in methanol, The selected ion transitions were m/z 415.2 → 258.2 and m/z 398.2 → 258.2 for SHR0302 and SHR143181 (internal standard), respectively. A full validation, including selectivity, linearity, carryover, precision, accuracy, recovery, matrix effect, dilution integrity and stability, was carried out in human plasma. It was successfully applied to a pharmacokinetic study in Chinese healthy subjects after oral administration of SHR0302 tablet., (© 2022 John Wiley & Sons Ltd.)

A rapid, simple, and economical method has been developed to determine colchicine in both human whole blood and urine using UPLC-MS/MS. Colchicine and isotope-labeled internal standard were extracted from the matrix by liquid-liquid extraction with saturated borax and ethyl acetate, and separated by a reversed-phase chromatography C18 column. Gradient elution was carried out using acetonitrile and water spiked with 0.01% formic acid. Multiple reaction monitoring was performed at positive ion mode. The quantitative transitions were m/z 400.27 → 310.28 for colchicine and m/z 406.16 → 313.18 for colchicine-D6. The method has good linearity in the range of 0.5-200 ng/mL for blood and 2-2000 ng/mL for urine. The sensitivity, accuracy, and matrix effect were all in line with the guidelines of Food and Drug Administration and European Medicines Agency. The extraction recovery was above 63.94%. The samples were stable under various storage conditions. Six deuterium-substituted isotopic internal standard was used to demonstrate a different mode of colchicine cleavage from the existing literature. This method has been successfully used in the diagnosis of patients with colchicine poisoning. Blood is recommended as the optimal sample compared with urine., (© 2022 John Wiley & Sons Ltd.)

Hederasaponin C (HSC), one of the main components of Pulsatilla chinensis, is considered as a potential drug for the treatment of inflammatory bowel disease. In the present research, we developed a pharmacokinetics-pharmacodynamics model to describe the concentration-effect course of drug action following the intraperitoneal injection of HSC in colitis rats. A sensitive UPLC-MS/MS method was established for the the determination of HSC in rat plasma to explore the pharmacokinetics properties. The separation was performed on an Accucore C 18 column (2.1 × 100 mm, 2.6 μm) with a flow phase consisting of acetonitrile and 0.1% formic acid water. The assay method was validated and demonstrated good adaptability for application in the pharmacokinetics study. Then the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) in colon tissues were measured using an ELISA assay. The levels of TNF-α, IL-1β and IL-6 were decreased after HSC administration, suggesting that HSC can significantly improve the level of inflammatory syndrome factor. The pharmacokinetics study showed that the time to peak concentration of HSC was 1 h. The concentration-effect curves showed a hysteresis loop. There was also a hysteresis between the peaked concentration and the maximum effect of HSC. The present study established in vivo pharmacokinetics-pharmacodynamics models and the results showed a great potential of HSC for treating ulcerative colitis., (© 2022 John Wiley & Sons Ltd.)

This research developed and validated a highly sensitive and selective UPLC-MS/MS approach using a triple quadrupole mass spectrometer for quantifying favipiravir. Moreover, we introduced a study evaluating bioequivalence using two drugs, Favibrivix and Avigan, containing favipiravir. Lean Six Sigma verified the capacity and performance of the process. Protein precipitation extraction was utilized to extract favipiravir from the collected human matrices. We used an Acquity UPLCr BEH HILIC column and valproic acid as an internal standard. Furthermore, we conducted the procedure using an isocratic elution comprising acetonitrile and 0.005% ammonia in water (75:25, v/v), a flow rate of 0.25 ml/min, a temperature controlled at 10°C and an injection volume of 1.0 μl. Our UPLC-MS/MS process has a broad range (50-10,000 ng/ml) with a determination coefficient of 0.9980. We validated the method in line with the US Food and Drug Administration. The findings revealed that the test, Favibrivix 200 mg/tablet, and the reference, Avigan® 200 mg/tablet, were statistically bioequivalent in healthy Egyptian participants., (© 2022 John Wiley & Sons, Ltd.)

A reliable and robust bioanalytical method was developed to quantify neratinib, a tyrosine kinase inhibitor in human plasma, using UPLC-MS/MS. The extraction of neratinib and its deuterated internal standard, neratinib-d6, was successfully performed on hybrid solid-phase extraction ultra-cartridges to remove the interference of phospholipids and proteins. Chromatographic analysis was performed on a UPLC BEH C18 (50 × 2.1 mm, 1.7 μm) column using 0.1% formic acid and acetonitrile under gradient conditions. The total analysis time was 1.5 min. Neratinib was quantified using electrospray ionization source operated in the positive-ion multiple reaction monitoring mode. The mass transitions of neratinib and neratinib-d6 were m/z 557.3/112.1 and m/z 563.1/118.2, respectively. The linear concentration range for neratinib was 0.5-500 ng/mL, which adequately covers concentration levels expected in real subject samples. The assay was extensively validated for various validation parameters following standard guidelines for a bioanalytical assay. The intra- and inter-batch precision was ≤4.6%, and neratinib was found to be stable under various stability conditions. The mean internal standard-normalized matrix factor and recovery were 0.997 and 95.4%, respectively. The validated method was successfully applied to a pharmacokinetic study in healthy subjects with different doses., (© 2022 John Wiley & Sons Ltd.)

This study developed a UPLC-MS/MS method to detect isoscoparin in mouse blood, determined the pharmacokinetics of isoscoparin in mice after intravenous (5 mg/kg) and intragastric (20 mg/kg) administration, and calculated the absolute bioavailability. A HSS T3 column was used for separation, and the column temperature was set at 40°C. The mobile phases were acetonitrile and 0.1% formic acid, and the gradient elution procedure was used. The blood sample was treated with protein precipitant with acetonitrile-methanol (9:1, v/v). Multiple reaction monitoring mode was used for quantitative analysis in electrospray positive-ion mode. It showed a good linear relationship in the range of 1-4000 ng/mL (r > 0.998); the intra-day and inter-day precision was <12%, and the accuracy was 86-112%. The recovery was >68%, and the matrix effect was 86-90%. The half-life of isoscoparin was relatively short in mice, and the bioavailability was 2.6%. The developed UPLC-MS/MS method was rapid, sensitive, and suitable for the pharmacokinetics of isoscoparin in mice., (© 2022 John Wiley & Sons Ltd.)

Introduction: Actinidia eriantha Benth radix (AEBR) is one of the most commonly used medicines by the She people in China, used primarily for the treatment of tumours of the digestive tract. There is currently limited to no data on the quality control of AEBR., Objectives: The aim of this study was to identify quality markers of AEBR., Material and Methods: An ultra-performance lquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) method was used to identify and analyse the components of AEBR from water extracts. An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was also established for the simultaneous determination of 13 active components in the water extracts. The network pharmacology method was used to screen for quality markers of AEBR in gastric cancer., Results: This study tentatively identified 199 chemical constituents and isomers, including 67 pentacyclic triterpenoids, 20 flavonoids, 39 phenolic acids, 18 coumarins, and other compounds. The 13 active components in the water extracts were successfully determined using a validated UPLC-MS/MS method. Based on the network pharmacology method, four compounds were selected as quality markers of AEBR., Conclusion: This study provides an important reference for the quality control of AEBR. Chemical analysis combined with network pharmacology provides an effective strategy for the discovery of quality markers in traditional Chinese/herb medicine., (© 2022 John Wiley & Sons Ltd.)

Introduction: The root of Tetrastigma hemsleyanum (RTH) has been widely used as a folk medicine in China. Meanwhile, its stems (STH) and leaves (LTH) are consumed as functional tea and food supplementation. Therefore, it is important to get a better understanding of the distribution of bioactive constituents in different parts of T. hemsleyanum., Objective: To develop a method for quantitative analysis of multiple bioactive constituents and comparing their distribution in RTH, STH and LTH., Methods: Ultra-performance liquid chromatography triple quadrupole ion trap tandem mass spectrometry (UPLC-QTRAP-MS/MS) was used for the quantitative analysis. The quantitative data were further analysed by principal component analysis (PCA), hierarchical cluster analysis (HCA) and partial least squares determinant analysis (PLS-DA)., Results: Forty-two constituents in RTH, STH and LTH, including 14 flavonoids, three phenolic acids, 15 amino acids and 10 nucleosides, were quantitatively determined. The contents of flavonoids and phenolic acids in LTH were significantly higher than those in RTH and STH. While the contents of amino acids and nucleosides in LTH were less than those in RTH and STH. Multivariate statistical analysis can significantly classify and distinguish RTH, STH, and LTH., Conclusions: The present method would be helpful for the quality control of T. hemsleyanum, and the results would be useful for the efficient utilisation of T. hemsleyanum in the future., (© 2022 John Wiley & Sons, Ltd.)

Dabigatran etexilate mesylate (DABE), a prodrug, quickly changes into dabigatran (DAB) after its oral administration. Accordingly, detecting DABE in plasma is practically unmanageable. An ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) technique was developed and validated to compute free DAB in participants. For the first time, the central composite design, a type of response surface methodology, was applied for optimizing variables affecting the cleavage of glucuronide bond. In addition, the pharmacokinetic parameters of generic medication (okanadab) were determined, and the obtained outcomes were compared with those of the branded drug (pradaxa). The sample preparation was done using methanol as a protein precipitant and the separation was achieved using an ACQUITY UPLC BEH C 18 column (2.1 × 50 mm, 1.7 μm). The elution was isocratically conducted using 10 mM ammonium formate:methanol (72:28, v/v) as a mobile phase and the flow rate was 0.25 mL/min. Multiple reaction monitoring and positive electrospray ionization were used. The determination was performed within 1 min, and the calibration growth curve was established over a range of 1.19-475 ng/mL using DAB-d3 as a tagged internal standard. Bioequivalence research was validated following the US Food and Drug Administration (US FDA) guidelines for bioanalytical procedures and acceptable outcomes were achieved. The outcomes for okanadab and pradaxa did not differ significantly., (© 2022 John Wiley & Sons, Ltd.)

Dyes on ancient silks have been a worth studying field through human's history, although current reports ignore the connection between natural dyes origin and relevant colour reduction methods, which poses an insurmountable obstacle for restoration of historical silks. In this paper, a series of 12 red hue silks from six natural dyes (sappanwood, Chinese madder, safflower, lac, cochineal, dragon's blood) via three different dyeing techniques were used to establish a self-built precise tandem mass spectrometry (MS/MS) database. With organic solvent extracting on those manual-dyed silks, ultraperformance liquid chromatography - electrospray ionization - quadrupole time of flight mass spectrometry (UPLC-ESI-QTOF-MS) was utilized to form preliminary MS database for screening and identifying of the potential dyes compounds without standard references. Furthermore, combining the targeted MS/MS mode and the matching threshold of 70.00, a self-built secondary MS/MS database was successfully established, which contains 33 compounds, 32 chromatograms and 32 MS/MS fragments. As for real sample application, the self-built precise MS/MS database had revealed that the dyes on two historical silks (Shanghai Museum, China) belong to Chinese madder just with different mordant dyeing ordinal. Additionally, by experimental restoration, visually indistinguishable silks (ΔE ab * < 1.5 NBS) were successfully restored. This explorative methodology can further inspire the traceability of biological dyestuffs, which lays instructive foundation on protection and restoration of artefacts, connecting the archaeological science and human art., (© 2022 John Wiley & Sons, Ltd.)

Rivaroxaban is an oral anticoagulant directly inhibiting the activity of Factor Xa, which is widely used for the prophylaxis of thromboembolic disorders. Therapeutic drug monitoring (TDM) is required during therapy for individual dosage adjustment. This study aimed at developing a liquid chromatography/tandem mass spectrometry method that was suitable for rivaroxaban TDM in human plasma and urine and exploring the feasibility of urine drug monitoring in medical care. A 3 min run time of the LC-MS/MS methods was established by employing an Acquity UPLC BEH C 18 (2.1 × 50 mm, 1.7 μm) column using gradient elution of 10 mmol/L ammonium acetate containing 0.1% formic acid-0.1% formic acid acetonitrile as a mobile phase at a flow rate of 0.4 ml/min with calibration ranges of 0.5-400 and 10-10,000 ng/ml for human plasma and urine, respectively. Rivaroxaban was detected on a triple quadrupole tandem mass spectrometer with an electrospray ionization source in positive ion mode. The methods showed good linearity within the calibration range. The precision and accuracy, matrix effect, extraction recovery and stability in both human matrices were all validated and meet the international guideline requirements. These validated methods were successfully applied to support the TDM of an aged patient receiving rivaroxaban for therapy., (© 2021 John Wiley & Sons, Ltd.)

Wu-tou decoction has been used as a traditional Chinese medicine prescription for thousands of years. It comprises five herbs, namely Radix Aconiti Preparata, Ephedrae Herba, Astragali Radix, Glycyrrhiza Radix, and Paeoniae Radix Alba. In addition, the original prescription contains honey, but in modern research, the existence of honey is commonly ignored. The aim was to investigate the effect of absorption in rats after oral wu-tou decoction with or without honey. In this research, a rapid and sensitive UPLC-MS/MS method was investigated for the quantitative analysis of ephedrine, pseudoephedrine, paeoniflorin, calycosin-7-glucoside, glycyrrhizic acid, liquiritin, and benzoylmesaconine in rat plasma after single and continuous oral decoctions. The results of the pharmacokinetic parameters showed that C max , CL/F, AUC 0-t , and AUC 0-∞ in the honey group were significantly increased than those in the non-honey group except for ephedrine and pseudoephedrine. The same trend was observed regardless of single or continuous oral administrations. Research studies showed that honey could promote the absorption of some effective components in wu-tou decoction in rats, enhance bioavailability, and provide a theoretical basis for the scientific and rational compatibility of the original prescription., (© 2022 John Wiley & Sons, Ltd.)

Trans-ferulic acid-4-β-glucoside (FAG) is a monomer extracted from Radix Aconiti Lateralis Preparata, which is a potential candidate for the prevention and treatment of cold injury. To determine the concentration of FAG in rats, it is essential to develop an ultra-performance liquid chromatography coupled with MS/MS method. Chromatographic separation was achieved by an Acquity UPLC BEH C18 column (2.1 × 50 mm, 1.7 μm). A Xevo triple quadrupole tandem mass spectrometer was used to quantitatively determine FAG in the negative-ion mode. The standard calibration curve was linear over the concentration range of 0.1-100 μg/mL and 0.0626-31.28 μg/g for rat plasma and liver tissue homogenate samples, respectively. The inter- and intra-batch precision (% relative standard deviation) of the assay was ≤8.29%, and accuracy (% relative error) ranged from -7.41 to 10.99%. The matrix effect was between 92.99 and 102.39%. The oral absolute bioavailability of FAG was obtained as 1.80%. The results of tissue distribution suggested that FAG spread rarely in the liver and brown adipose, which was not propitious to exert its ability to treat cold injury. In general, these studies were significant to provide necessary information for further study., (© 2022 John Wiley & Sons, Ltd.)