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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. In this study, a multi‐component determination method for the active ingredients was established, and the differences of active ingredients content in GBE was uncovered. Moreover, the possible key components of GBE anti‐IS were identified by content analysis and network pharmacology. The feasibility of GBE for the treatment of IS was confirmed from an anti‐inflammatory perspective and the key role of regulated lipid and atherosclerosis, PI3K‐Akt signaling pathway, and pathways of neurodegeneration. This study provides new strategy into mechanisms of GBE anti‐IS. [ABSTRACT FROM AUTHOR]

Copyright of Traditional Chinese Drug Research & Clinical Pharmacology is the property of Traditional Chinese Drug Research & Clinical Pharmacology Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective: A remarkably sensitive, accurate, and efficient ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) approach was developed as a facile and expeditious method for measuring cilofexor concentration in beagle dogs, the herb-drug interactions between silybinin and cilofexor was explored based on pharmacokinetics. Methods: The plasma sample protein of the beagles were rapidly sedimented with acetonitrile, and cilofexor and tropifexor (internal standard, ISTD) were separated by gradient elution using a 0.1% formic acid aqueous solution and acetonitrile as the mobile phase. The concentrations were detected using positive ion multiple reaction monitoring (MRM) mode. Mass transfer pairs were m/z 587.91-267.91 for cilofexor and m/z 604.08228.03 for ISTD, respectively. A two-period self-controlled experimental design was adopted for the HDIs experiment. In the first period (Group A), six beagle dogs were orally administered cilofexor at a dose of 1 mg/kg. In the second period (Group B), silybinin (3 mg/kg) was orally administered to the six beagle dogs twice a day for seven consecutive days, after which cilofexor was orally administered. The cilofexor concentration in beagle dogs was determined, and HDIs were evaluated based on their pharmacokinetics. Results: The accuracy and precision of cilofexor were both less than 15%, and the recoveries, matrix effects, and stability met the relevant requirements. The Cmax of cilofexor in group B was 49.62% higher than that in group A, whereas the AUC(0-t) and AUC(0-8) of cilofexor in group B were 47.85% and 48.52% higher, respectively, than those in group A. Meanwhile, the t1/2 extended from 7.84 h to 9.45 h, CL and Vz decreased in Group B. Conclusion: A novel UPLC-MS/MS approach was successfully applied for the measurement of cilofexor in beagle dog plasma. Silybinin can alter the pharmacokinetics of cilofexor in beagle dogs, thereby increasing plasma exposure to cilofexor. [ABSTRACT FROM AUTHOR]

Thymus species (Lamiaceae), commonly referred to as thyme, have gained popularity due to their utilisation in traditional medicine for various ailments such as gastrointestinal disorders, colds, upper respiratory infections, vomiting, jaundice, oral hygiene and oral ulcers. Nevertheless, a significant number of Thymus species remain unexplored in terms of their bioactivity and phenolic content. In the present study, a thorough chemical analysis of endemic Thymus canoviridis methanol (METC)-water extracts (WETC) and Thymus pubescens var. pubescens methanol (METP)-water extracts (WETP) was carried out by Ultra-Performance Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS/MS). In addition, the inhibitory effect and antioxidant properties of these extracts on the enzymes acetylcholinesterase (AChE), α -glycosidase and carbonic anhydrase II (CA II) were also evaluated. The antioxidant potential was assessed using six assays, namely DPPH, ABTS, DMPD, FRAP, CUPRAC and Fe3+ reducing assays. Our results suggest that the methanol extract from these two plants exhibits a higher level of antioxidant activity. Furthermore, WETP inhibited all enzymes in the best way. According to the UPLC-MS/MS analysis results, rosmarinic acid showed its highest content in METC (18285.55 µg/g extract), WETC (27.98 µg/g extract) and METP (2910.59 µg/g extract), while WETP (69.76 µg/g extract) and the following total amounts of phenolic compounds were METC 20726.55 µg/g extract, METP 5887.20 µg/g extract, WETP 348.98 µg/g extract, WETC 68.56 µg/g extract. It is consistent with the analytical result, total phenolic content, and activity ranking in antioxidant assays. Regard to the results of the molecular docking studies, there is a high binding affinity between the CA II enzyme and cyanidin-3- O -glycoside, between the AChE and keracyanin chloride, and between α -glycosidase enzyme and rosmarinic acid. The extracts were discovered to exhibit various biological activities, such as antioxidant, antidiabetic, anti-Alzheimer's disease, and antiglaucoma properties. [Display omitted] • Antioxidant and antimicrobial activities of Thymus species were evaluated using several bioassays. • Comprehensive metabolic profiling of Thymus species was relaized by UPLC-MS/MS. • Anti-Alzheimer, antidiabetic, anti-glaucoma effects of Thymus species were determined and compared to standards. • High binding affinity was detected between CA II enzyme-cyanidin-3- O -glucoside, AChE-keracyanin chloride, and α -glycosidase-rosmarinic acid. [ABSTRACT FROM AUTHOR]

Per- and poly-fluoroalkyl substances (PFAS) are a large group of man-made chemicals that repel oil, stains, grease, and water and are fire resistant. PFAS are known to be persistent in the environment and bioaccumulate in human and animal tissue, soil, and water. To mitigate human exposure to these chemicals, PFAS analysis by ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) was used to monitor their presence in the environment. This study focused on analyzing oysters harvested from coastal New England for 14 PFAS. The validated method addressed in this study utilized a quick, easy, cheap, effective, rugged, and safe (QuEChERS) approach using various sorbents and showed that the combination of primary secondary amine/graphitized carbon black (PSA/GCB) provided the most effective sample clean-up prior to analysis. The recoveries for the targeted analysis ranged from 61% to 116.3% with relative standard deviations from 2.4% to 13.3% at the 125.0 ng mL−1 analyte level. The method detection limits were from 0.33 ng g−1 to 6.75 ng g−1. In a targeted analysis of five unknown samples, two samples had a detectable level of the legacy compound, perfluorooctanesulfonic acid (PFOS). [ABSTRACT FROM AUTHOR]

Aim: Our study is to investigate the effects of triazole antifungal drugs on the pharmacokinetics of lorlatinib in rats. Methods: The samples were precipitated with methanol. Chromatographic separation was performed on a ultra-performance liquid chromatography (UPLC) system using a BEH C18 column. The mobile phase consisted of 0.1% formic acid water and methanol. Lorlatinib and crizotinib (internal standard) were detected in multiple reaction monitoring mode. The fragment ions were 407.3–228.07 for lorlatinib and m/z 450.3–260.0 for crizotinib. Lorlatinib and different triazole antifungal drugs were given to Sprague Dawley rats by gavage, and blood was collected from the tail vein at a certain time point. The validated UPLC–MS/MS method was applied to a drug interaction study of ketoconazole, voriconazole, itraconazole, and posaconazole with lorlatinib in rats. Results: Ketoconazole and voriconazole significantly inhibited lorlatinib metabolism. When administration with ketoconazole and voriconazole, the area under the curve from time zero to infinity of lorlatinib increased by 49.0% and 104.3%, respectively; the clearance decreased by 40.0% and 40.0%, respectively. While itraconazole and posaconazole did not affect lorlatinib pharmacokinetics. Conclusion: The UPLC-MS/MS-based assay is helpful to further understand the pharmacokinetics of lorlatinib in rats, and confirmed the findings that the combination of lorlatinib with CYP3A inhibitors should be avoided as predicted by our pre-clinical studies. [ABSTRACT FROM AUTHOR]

To distinguish the diflerences in the types and contents of flavonol glycoside in green tea from 6 production areas, and explore the feasibility of flavonol glycoside for geographical traceability of green tea. In this study, the qualitative identification of flavonol glycosides by ultrahigh performance liquid chromatography-quadrupole-electrostatic field or- bitrap high resolution mass spectrometry (UPLC-Q-Orbitrap HRMS), and a quantitative method of flavonol glycosides in green tea was established based on ultra high performance liquid chromatography -tandem triple quadrupole mass spec- trometry (UPLC-MS/MS). The results showed that a total of 21 kinds of flavonol glycosides were positively identified in green tea, including 5 myricetin glycosides, 8 quercetin glycosides and 8 kaempferol glycosides. The quantitative analysis showed that the content of flavonol glycosides varied from different producing area in green tea, in which the content of K-glu-rha-glu was the highest, accounting for 33.1% -57.8% of 21 kinds of flavonol glycosides, followed with the kaempferol glycoside, while content of myricetin glycoside was the lowest. By analyzing the differences and correlations of flavonol glycosides in green tea from 6 production areas indicate that they can be used for geographical traceability of tea. The green teas from different producing areas could be effectively distinguished by PCA. The precision rate was 100.0% for BP-ANN recognition, and 2 kinds of compound markers for flavonol glycoside were acquired with 100% importance (G<0.05). The predictive ability of PLS-DA model was 78.2%, and 8 biomarkers (VIP>1) were screened out. The entirety precision rate was 100% by LSD model, and the precision rate of cross validation result was 100%. The results of this study may provide scientific basis for the study of geographical traceability and chemical classification of tea. [ABSTRACT FROM AUTHOR]

An effective analytical method using NTM-COFs on-line pass-through cleanup method and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to simultaneously quantify 12 illicit drugs (norketamine, ketamine, codeine, cocaine, methamphetamine, 3,4-methylenedioxyamphetamine, 3,4-methylenedioxymethamphetamine, amphetamine, benzoylecgonine, 6-acetylmorphine, morphine and methadone) in hair sample was developed and validated. The hair sample was extracted with 0.3% formic acid-acetonitrile (v/v) and passed through the nano-titania-modified covalent organic frameworks (NTM-COFs) pre-column to reduce the matrix effect caused by hair specimen. Under the optimal conditions, satisfactory extraction efficiency was obtained for 12 illicit drugs with recoveries between 84.6% and 108.8%. The proposed method was linear for each analyte with correlation coefficients over 0.9926. The limits of detection and limits of quantification for 12 illicit drugs were in the range of 0.012–0.558 ng/g and 0.036–1.860 ng/g, respectively. Acceptable precision and accuracy were also achieved with relative standard deviations values below 9.92% and relative error values below 15.4%. The analytical method was successfully applied to real hair samples from suspected drug abusers, which was proved to be reliable and robust for drug screening in forensic toxicology. [ABSTRACT FROM AUTHOR]

Sea buckthorn pomace (SBP) is a by-product of sea buckthorn processing that is rich in bioactive compounds. In this study, different active ingredients were extracted by using different solvents (water, methanol, ethanol, glycerol, ethyl acetate, and petroleum ether) combined with an ultrasonic assisted method. The correlation between the active ingredients and antioxidant properties of the extract was studied, which provided a research basis for the comprehensive utilization of SBP. This study revealed that the 75% ethanol extract had the highest total phenolic content (TPC) of 42.86 ± 0.73 mg GAE/g, while the 75% glycerol extract had the highest total flavonoid content (TFC) of 25.52 ± 1.35 mg RTE/g. The ethanol extract exhibited the strongest antioxidant activity at the same concentration compared with other solvents. The antioxidant activity of the ethanol, methanol, and glycerol extracts increased in a concentration-dependent manner. Thirteen phenolic compounds were detected in the SBP extracts using UPLC-MS/MS analysis. Notably, the 75% glycerol extract contained the highest concentration of all identified phenolic compounds, with rutin (192.21 ± 8.19 μg/g), epigallocatechin (105.49 ± 0.69 μg/g), and protocatechuic acid (27.9 ± 2.38 μg/g) being the most abundant. Flavonols were found to be the main phenolic substances in SBP. A strong correlation was observed between TPC and the antioxidant activities of SBP extracts. In conclusion, the choice of solvent significantly influences the active compounds and antioxidant activities of SBP extracts. SBP extracts are a valuable source of natural phenolics and antioxidants. [ABSTRACT FROM AUTHOR]

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‐d6 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. [ABSTRACT FROM AUTHOR]

Copyright of Food & Fermentation Industries is the property of Food & Fermentation Industries and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

DAPB, a new molecule including danshensu, borneol, and a mother nucleus of ACEI (Angiotensin-converting enzyme inhibitors), is being developed as an antihypertensive candidate compound. A rapid, accurate, and sensitive ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method was established and validated for the determination of DAPB in rat plasma. Chromatographic separation was performed on an Agilent SB-C18 column after protein precipitation by acetonitrile with a mobile phase consisting of acetonitrile and deionized water with 0.02% formic acid and 5 mM NH4F (v/v) at a flow rate of 0.2 mL/min. Quantification was performed using electrospray positive ionization mass spectrometry in the multiple reaction monitoring (MRM) mode. The method was linear over the range of 2–1000 ng/mL. The intra- and inter-day precision was within 12%, with accuracies less than 7%. Stability was within the acceptable limits under various storage and processing conditions. No apparent matrix effect was detected. The validated method was applied to the pre-clinical pharmacokinetic study of DAPB after oral administration of 30 mg/kg and intravenous administration of 6 mg/kg in rats. [ABSTRACT FROM AUTHOR]

Ginseng Radix et Rhizoma Rubra (Panax ginseng C.A. Mey, Hongshen, in Chinese) and Ophiopogonis Radix (Ophiopogon japonicus (L.f) Ker-Gawl., Maidong, in Chinese) are traditional Chinese herbal pairs, which were clinically employed to enhance the immune system of cancer patients. This study employed the pharmacokinetic and pharmacodynamic (PK–PD) spectrum-effect association model to investigate the antitumor active substances of P. ginseng and O. japonicus (PG–OJ). The metabolic processes of 20 major bioactive components were analyzed using Ultra-Performance Liquid Chromatography–Mass Spectrometry/Mass Spectrometry (UPLC–MS/MS) in the lung tissue of tumor-bearing mice treated with PG–OJ. The ELISA method was employed to detect the levels of TGF-β1, TNF-α, and IFN-γ in the lung tissue of mice at various time points, and to analyze their changes after drug administration. The results showed that all components presented a multiple peaks absorption pattern within 0.083 to 24 h post-drug administration. The tumor inhibition rate of tumor and repair rate of IFN-γ, TNF-α, and TGF-β1 all increased, indicating a positive therapeutic effect of PG–OJ on A549 tumor-bearing mice. Finally, a PK–PD model based on the GBDT algorithm was developed for the first time to speculate that Methylophiopogonanone A, Methylophiopogonanone B, Ginsenoside Rb1, and Notoginsenoside R1 are the main active components in PG–OJ for lung cancer treatment. [ABSTRACT FROM AUTHOR]

This study delved into the impact of two extrusion processing parameters—screw speed (SS at 400, 600, 800 RPM) and material moisture content in the extruder barrel (M at 12, 15, 18%) at constant feed rate (50 kg/h)—on reducing the content of alternariol (AOH), alternariol monomethyl ether (AME), tenuazonic acid (TeA), and tentoxin (TEN) in whole-grain red sorghum flour. Ultra-performance liquid chromatography combined with a triple-quadrupole mass spectrometer (UPLC-MS/MS) was employed for the determination of Alternaria toxin levels. The extruder die temperature fluctuated between 136 and 177 °C, with die pressures ranging from 0.16 to 6.23 MPa. The specific mechanical energy spanned from 83.5 to 152.3 kWh/t, the torque varied between 88 and 162.8 Nm, and the average material retention time in the barrel ranged from 5.6 to 13 s. The optimal parameters for reducing the concentration of all Alternaria toxins with a satisfactory quality of the sorghum snacks were: SS = 400 RPM, M = 12%, with a reduction of 61.4, 76.4, 12.1, and 50.8% for AOH, AME, TeA, and TEN, respectively. [ABSTRACT FROM AUTHOR]

This study aims to explore the effects of frying conditions on the formation of HAs and PAHs in crispy pork spareribs, a popular meat commodity sold on Taiwan's market. Raw pork spareribs were marinated, coated with sweet potato powder, and fried in soybean oil and palm oil at 190 °C/6 min or 150 °C/12 min, followed by an analysis of HAs and PAHs via QuEChERS coupled with UPLC-MS/MS and GC-MS/MS, respectively. Both HAs and PAHs in pork spareribs during frying followed a temperature- and time-dependent rise. A total of 7 HAs (20.34–25.97 μg/kg) and 12 PAHs (67.69–85.10 μg/kg) were detected in pork spareribs fried in soybean oil and palm oil at 150 °C/12 min or 190 °C/6 min, with palm oil producing a higher level of total HAs and a lower level of total PAHs than soybean oil. The content changes of amino acid, reducing sugar, and creatinine played a vital role in affecting HA formation, while the degree of oil unsaturation and the contents of precursors including benzaldehyde, 2-cyclohexene-1-one, and trans,trans-2,4-decadienal showed a crucial role in affecting PAH formation. The principal component analysis revealed that HAs and PAHs were formed by different mechanisms, with the latter being more liable to formation in pork spareribs during frying, while the two-factorial analysis indicated that the interaction between oil type and frying condition was insignificant for HAs and PAHs generated in crispy pork spareribs. Both CcdP (22.67–32.78 μg/kg) and Pyr (16.70–22.36 μg/kg) dominated in PAH formation, while Harman (14.46–17.91 μg/kg) and Norharman (3.41–4.55 μg/kg) dominated in HA formation in crispy pork spareribs during frying. The outcome of this study forms a basis for learning both the variety and content of HAs and PAHs generated during the frying of pork spareribs and the optimum frying condition to minimize their formation. [ABSTRACT FROM AUTHOR]

Aim: The study we are conducting aims to establish a validated method for measuring empagliflozin in plasma using UPLC MS/MS. Materials and Methods: Sodium-Glucose co Transporter inhibitors (SGLTi), a novel class of diabetes medication. Non-insulin dependent diabetes and adult-onset diabetes were the prior names for type 2 diabetes. Empagliflozin was the first SGLT2-inhibitor a drug used that reduced the risk of cardiovascular risk those with type 2 diabetes and pre-existing cardiovascular disease. The empagliflozin D4 as Internal Solution (IS). Liquid-liquid extraction was used to achieve separation on Synergi 2.5µ Fusion-Reverse phase 100A (100 mm×2.0 mm), 2.5 µm column. Mixture of Methanol: As a mobile phase, 0.2% formic acid in a 75:25 volumetric ratio is utilised. The flow rate is set at 0.3µl/min. The calibration curve is linear and plotted. Results: The developed method is accurate, precise, sensitive method for determination of empagliflozin from human plasma solution. Ethyl acetate and water in HPLC grade are preferred solvent for empagliflozin. Performing LLE extraction with centrifuge and LV Nitrogen Evaporator it with Ethyl acetate and water. Ethyl acetate is used as non-polar solvent and water is used as polar solvent for liquid-liquid extraction. LLE is a low cost extraction process compare to solid liquid extraction. The reported method is suitable for bioequivalence and pharmacokinetic studies. [ABSTRACT FROM AUTHOR]

Coffee cherries contain a crucial flavor-precursor and chemical substances influencing roasted bean quality, yet limited knowledge exists on metabolite changes during cherry ripening. Our study identified 1078 metabolites, revealing 46 core differential metabolites using a KEGG pathway analysis. At the GF vs. ROF stage, amino acid synthesis dominated; ROF vs. BRF featured nucleotide catabolism; BRF vs. PRF exhibited glycoside and flavonoid synthesis; and PRF vs. PBF involved secondary metabolite synthesis and catabolism. The PRF stage emerged as the optimal cherry-harvesting period. A correlation analysis identified core differential metabolites strongly linked to taste indicators, suggesting their potential as taste markers. Notably, nucleotides and derivatives exhibited significant negative correlations with glycosides and flavonoids during ripening. This research systematically analyzed flavor and active substances in green coffee beans during cherry ripening, offering valuable insights into substance formation in Coffea arabica L. [ABSTRACT FROM AUTHOR]

A method employing online solid phase extraction (SPE) coupled to UPLC-MS/MS was developed for the determination of residues of the acid herbicide quinclorac plus its transformation product, quinclorac methyl ester, in honey. The analytical method involved dissolving the honey in a mixture of methanol:water followed by direct injection into a two-dimensional UPLC system which is used to perform an automated SPE cleanup on a reusable phenyl cartridge prior to the target analytes being transferred onto an analytical UPLC column for subsequent chromatographic separation followed by MS/MS detection. The limits of quantitation for quinclorac and quinclorac methyl ester in honey were both set at 0.5 µg kg−1 and the method detection limit was estimated to be 0.012 µg kg−1 for each compound. The working analytical range (0.5–100 µg kg−1) was validated by analysing a series of spiked replicate honey samples. The method was applied to the analysis of various honeys obtained from numerous different commercial sources. Quinclorac was detected in 9 out of 30 samples at concentrations ranging from 0.6 to 31.5 µg kg−1. Quinclorac methyl ester, which is estimated to be significantly more toxic than the parent herbicide itself, was not detected in any honey sample. [ABSTRACT FROM AUTHOR]

Gibberellin (GA) is an important plant hormone that is involved in various physiological processes during plant development. Sweet cherries planted in southern China have always encountered difficulty in bearing fruit. In recent years, gibberellin has successfully solved this problem, but there has also been an increase in malformed fruits. This study mainly explores the mechanism of malformed fruit formation in sweet cherries. By analyzing the synthesis pathway of gibberellin using metabolomics and transcriptomics, the relationship between gibberellin and the formation mechanism of deformed fruit was preliminarily determined. The results showed that the content of GA3 in malformed fruits was significantly higher than in normal fruits. The differentially expressed genes in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were mainly enriched in pathways such as "plant hormone signal transduction", "diterpenoid biosynthesis", and "carotenoid biosynthesis". Using Quantitative Real-Time Reverse Transcription PCR (qRT-PCR) analysis, the gibberellin hydrolase gene GA2ox and gibberellin synthase genes GA20ox and GA3ox were found to be significantly up-regulated. Therefore, we speculate that the formation of malformed fruits in sweet cherries may be related to the accumulation of GA3. This lays the foundation for further research on the mechanism of malformed sweet cherry fruits. [ABSTRACT FROM AUTHOR]

In this study, a ultra performance liquid chromatography-tandem mass spectrometry method for the determination of methyltestosterone in tilapia was optimized and established. Tilapia samples were homogenized, extracted by acetonitrile solution containing 1.0% formic acid, purified by QuEChERS, concentrated by nitrogen blowing in water bath, and dissolved in mobile phase. The sample was separated on a BEH C18 column and eluted with a gradient elution of aqueous solution containing 0.1% formic acid and methanol as mobile phase, using ESI positive ion multiple reaction monitoring mode in UPLC-MS/MS, and quantitative analysis was used with internal standard method. The results showed that methyltestosterone had a good linear relationship within the linear range. The correlation coefficient of the standard curve was 0.999 5. The detection limit was 0.30 μg/kg and the quantitative limit was 1.0 μg/kg. At the different spiked levels (n=6) in tilapia substrate, the average recoveries were 98.3% to 103.0% and the relative standard deviation was 4.2% to 8.8%. The optimized method has high sensitivity, good accuracy, simple and rapid pre-treatment process, which is suitable for the detection of methyltestosterone residues in tilapia. [ABSTRACT FROM AUTHOR]

Acylcarnitines are metabolic intermediates of fatty acids and branched-chain amino acids having vital biofunctions and pathophysiological significances. Here, we developed a high-throughput method for quantifying hundreds of acylcarnitines in one run using ultrahigh performance liquid chromatography and tandem mass spectrometry (UPLC-MS/MS). This enabled simultaneous quantification of 1136 acylcarnitines (C0–C26) within 10-min with good sensitivity (limit of detection < 0.7 fmol), linearity (correlation coefficient > 0.992), accuracy (relative error < 20%), precision (coefficient of variation (CV), CV < 15%), stability (CV < 15%), and inter-technician consistency (CV < 20%, n = 6). We also established a quantitative structure-retention relationship (goodness of fit > 0.998) for predicting retention time (t R) of acylcarnitines with no standards and built a database of their multiple reaction monitoring parameters (t R , ion-pairs, and collision energy). Furthermore, we quantified 514 acylcarnitines in human plasma and urine, mouse kidney, liver, heart, lung, and muscle. This provides a rapid method for quantifying acylcarnitines in multiple biological matrices. [Display omitted] • A sensitive 10-min method for simultaneous quantification of multiple acylcarnitines. • Quantification of 514 acylcarnitines in multiple biofluids and tissues for molecular phenotyping. • Quantitative structure-retention relationship and database of MRM parameters (t R , ion-pairs, and CE) for 1136 acylcarnitines. [ABSTRACT FROM AUTHOR]

Vancomycin is a critical antibiotic frequently utilized in clinical settings, with therapeutic drug monitoring (TDM) strongly advised to optimize treatment efficacy and mitigate the risk of adverse effects. However, current methods for measuring vancomycin levels in human plasma are hindered by long analysis times and complicated sample preparations. Thus, this study developed and validated a novel UPLC-MS/MS method for a rapid (with a running time of 3.5 min) and simple analysis of plasma vancomycin. To quantify vancomycin concentration in human plasma, we have developed and validated the UPLC-MS/MS method with high sensitivity, specificity, and accuracy, meeting the strict criteria according to the Food and Drugs Administration (FDA) guidelines for validation biological analysis methods. Vancomycin and atenolol (internal standard) underwent positive electrospray ionization (ESI+) and detection in multi-reaction monitoring (MRM) mode. The selected MRM transitions were m/z 725.66→144.16 for vancomycin and m/z 267.29→189.96 for atenolol. Plasma samples were precipitated using a simple mixture containing acetonitrile, methanol, and formic acid as a pH adjuster. The separation was performed using the Poroshell 120 Phenyl Hexyl Column (4.6 × 150 mm, 2.7 μm) maintained at 25 °C for 3.5 min. Isocratic elution with a mobile phase (methanol and 0.1% formic acid in a 40:60 v/v ratio) at a flow rate of 0.5 mL/min was employed. The method showed linearity (0.1-75 μg/mL) with a coefficient of determination above 0.9994 and a lower limit of quantification at 0.1 μg/mL. Precision, both intraday and interday, was below 10%, and accuracy ranged from 91.70% to 111.57%. System suitability, selectivity, stability, carryover, dilution, recovery, and matrix effect validation results all met acceptable criteria. The established UPLC-MS/MS method is expected to be a rapid, simple, and reliable tool for drug monitoring and pharmacokinetic studies, enhancing patient care during vancomycin administration. [ABSTRACT FROM AUTHOR]

Objective: This study sought to evaluate and validate a method for chemical composition analysis and content determination of Goupi plaster components, a conventional prescription preparation of traditional Chinese medicine. This is geared toward providing a basis for quality control research and future development of Goupi plaster. Methods: UPLC–Q-Exactive-MS was used to qualitatively analyze the chemical components of Goupi plaster from different manufacturers in positive and negative ion modes. UPLC–MS/MS method was used to establish the determination methods for the detection of sinomenine, osthole, and cinnamaldehyde in Goupi plaster from different manufacturers. Results: A total of 291 chemical components were identified in Goupi plaster from four manufacturers, including 97 chemical components with known source and pharmacological activity. Further, we determined the contents of sinomenine, osthole, and cinnamaldehyde. Conclusion: In summary, the UPLC–Q-Exactive-MS method was used to analyze the chemical components of Goupi plaster from different manufacturers. We established the UPLC–MS/MS method to determine the contents of sinomenine, osthole, and cinnamaldehyde in Goupi plaster from different manufacturers. The findings indicated that the method was comprehensive, rapid, and accurate, preliminarily revealing the material basis of Goupi plaster and providing a reference for follow-up development of Goupi plaster. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Bioprocess Engineering is the property of Chinese Journal of Bioprocess Engineering Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Alcohol abuse and addiction is a public health issue of global concern. Wastewater-based epidemiology (WBE) is a forceful and effective complementary tool for investigating chemical consumption. This study examined alcohol consumption in major cities of China via WBE and compared WBE estimates with other data sources. A simple and valid ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the determination of two alcohol metabolites, ethyl glucuronide (EtG) and ethyl sulfate (EtS) in wastewater. The optimized method was applied to 62 sewage samples collected from wastewater treatment plants (WWTPs) in 31 provincial capital cities across China in the fourth quarter of 2020. The methodology established in this study was validated with the lower limit of quantification (LLOQ) up to 0.1 μg/L, good linearity in the range of 0.1–50 μg/L, intra-day and inter-day precision less than 5.58% and 5.55%, respectively, and the recoveries of the extracts were higher than 97.14%. The consumption range of alcohol estimated via WBE was 6.09 ± 4.56 ethanol/person/day (EPD) in the capital cities of China. Alcohol consumption varies significantly between cities in China, with WBE estimating lower alcohol consumption than WHO and lower than foreign countries. Investing in alcohol consumption based on WBE has great potential to accurately and efficiently estimate alcohol consumption. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Copyright of Food & Machinery is the property of Food & Machinery Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

As a medicine-food homology herb, Dendrobium spp. has versatile applications in modern medicine and food industry. Herein, an ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) based method was established for simultaneous quantification of six active components, including gigantol, erianin, naringenin, quercetin, rutin, and p-coumaric acid in Dendrobium spp., on the basis of optimized sample preparation, mass spectrometry conditions, and chromatography conditions. Sample extraction was carried out using methanol at a temperature of 60 °C, followed by separation on a T3 C18 column utilizing a gradient eluting program. The results demonstrated excellent linearity (r > 0.999) for the six active components within a specified concentration range. The average recovery rates ranged from 84.7 % to 106.9 %, and the precision (RSD) was within 7.4 %. The detection and quantification limits of this method ranged from 0.34 to 4.17 ng mL−1 and 1.12–13.91 ng mL−1, respectively. The established method demonstrates high accuracy and reliability and is applicable in practical sample detection. Different Dendrobium spp. exhibit specific variations in compound composition, with D. fimbriatum Hook. having a higher content of benzyl compounds and D. crystallinum. Rchb. f. having a higher content of flavonoids. This study provides experimental evidence for the quality and safety regulation of Dendrobium spp.

In this study, a UPLC-MS/MS method was developed for the rapid detection of 71 neuropsychotropic drugs in human serum for drug concentration monitoring and toxicity screening. The analytes were separated from the biological matrix by protein precipitation using a methanol-acetonitrile solvent mixture. The chromatographic separation was performed on a Kromasil ClassicShell C18 column (2.1*50 mm, 2.5 μ m) with gradient elution using acetonitrile-0.2 % acetic acid and 10 mM ammonium acetate as the mobile phases (flow rate 0.4 mL/min, column temperature 40 °C, injection volume 5 μL). An electrospray ion source in both positive and negative ion modes with multiple ion monitoring was used. The total run time was 6 min. All compounds were quantified using the isotope internal standard method. Totally, 71 drugs were detected within their linear ranges with correlation coefficients greater than 0.990. The intra- and inter-batch precision relative standard deviations (RSDs) for the low, medium, and high concentration points were less than 15 %, with an accuracy of 90%–110 %. All compounds except Moclobemide N-oxindole are stabilised within 7 days. The relative matrix effect results for each analyte were within ±20 % of the requirements. The method is validated according to Clinical and Laboratory Standards Institute guidelines, easy to use, and has a low cost.

Yellow tea (YT) has an additional process of yellowing before or after rolling than green tea (GT), making YT sweeter. We analyzed the variations of composition and taste throughout the withering, fixing and rolling steps using UPLC-MS/MS and sensory evaluation, and investigated the influence of various yellowing times on flavor profile of YT. 532 non-volatile metabolites were identified. Withering and fixing were the important processes to form the taste quality of GT. Withering, fixing and yellowing were important processes to form flavor profile of YT. Withering mainly regulated bitterness and astringency, and fixing mainly regulated bitterness, astringency and sweetness of YT and GT. Yellowing mainly regulated sweetness of YT. Trans-4-hydroxy-L-proline and glutathione reduced form as the key characteristic components of YT, increased significantly during yellowing mainly through Arginine and proline metabolism and ABC transporters. The paper offers a systematic insight into intrinsic mechanisms of flavor formation in YT and GT.

PAXLOVID™ (Co-packaging of Nirmatrelvir with Ritonavir) has been approved for the treatment of Coronavirus Disease 2019 (COVID-19). The goal of the experiment was to create an accurate and straightforward analytical method using ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) to simultaneously quantify nirmatrelvir and ritonavir in rat plasma, and to investigate the pharmacokinetic profiles of these drugs in rats. After protein precipitation using acetonitrile, nirmatrelvir, ritonavir, and the internal standard (IS) lopinavir were separated using ultra performance liquid chromatography (UPLC). This separation was achieved with a mobile phase composed of acetonitrile and an aqueous solution of 0.1% formic acid, using a reversed-phase column with a binary gradient elution. Using multiple reaction monitoring (MRM) technology, the analytes were detected in the positive electrospray ionization mode. Favorable linearity was observed in the calibration range of 2.0–10000 ng/mL for nirmatrelvir and 1.0–5000 ng/mL for ritonavir, respectively, within plasma samples. The lower limits of quantification (LLOQ) attained were 2.0 ng/mL for nirmatrelvir and 1.0 ng/mL for ritonavir, respectively. Both drugs demonstrated inter-day and intra-day precision below 15%, with accuracies ranging from −7.6% to 13.2%. Analytes were extracted with recoveries higher than 90.7% and without significant matrix effects. Likewise, the stability was found to meet the requirements of the analytical method under different conditions. This UPLC-MS/MS method, characterized by enabling accurate and precise quantification of nirmatrelvir and ritonavir in plasma, was effectively utilized for in vivo pharmacokinetic studies in rats.

In order to investigate the dynamic changes of flavor compounds, Ultra Performance Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS/MS) combined with Headspace Solid Phase Microextraction Gas Chromatography Mass Spectrometry (HS-SPME-GC–MS) was used to detect the metabolites in different drying processes. A total of 80 volatile compounds and 1319 non-volatile compounds were identified. The trend in the changes of C-8 compounds and sulfur-containing compounds were generally consistent with the trend of key enzyme activities. 479 differential metabolites were identified and revealed that metabolic profiles of compounds in Boletus edulis were altered with increased organic acids and derivatives and lipids and lipid-like molecules. Fatty acids and amino acids were transformed into volatile compounds under the action of enzymes, which played a significant role in the formation of the distinctive flavor of Boletus edulis. Our study provided a theoretical support for fully comprehending the formation mechanism of flavor from Boletus edulis during drying processes.

Background: Constipation is one of the chronic gastrointestinal functional diseases that affects the quality of life. While Qi Lang Formula (QLF) has demonstrated effectiveness in alleviating constipation symptoms, its precise mechanism remains elusive. Methods: QLF was analyzed using UPLC-MS/MS. Targets for QLF were collected from SwissADME, Herb, ITCM databases, and constipation-related targets from scRNA-seq and Genecards databases. Overlapping targets suggested potential QLF therapy targets for constipation. Enrichment analysis used the KOBAS database. A ''drug-ingredient-target'' network was constructed with Cytoscape, and AutoDock verified active ingredient binding. H&E staining assessed colonic mucosa changes, TEM examined ICC structural changes. ELISA measured neurotransmitter levels, and Western blot verified QLF's effect on target proteins. ICC proliferation was observed through immunofluorescence. Results: We identified 89 targets of QLF associated with ICC-related constipation, with c-Kit emerging as the pivotal target. Molecular docking studies revealed that Atractylenolide Ⅲ, Apigenin, Formononetin, Isorhamnetin, Naringenin, and Ononin exhibited strong binding affinities for the c-Kit structural domain. QLF significantly enhanced first stool passage time, fecal frequency, fecal moisture content, and intestinal propulsion rate. Further analysis unveiled that QLF not only restored neurotransmitter levels but also mitigated colon muscular fiber ruptures. ICC ultrastructure exhibited partial recovery, while Western blot confirmed upregulated c-Kit expression and downstream targets. Immunofluorescence results indicated ICC proliferation post QLF treatment in rat colon. Conclusion: Our findings suggest that QLF may promote ICC proliferation by targeting SCF/c-Kit and its downstream signaling pathway, thereby regulating intestinal motility.

Antibiotics and sedatives are used in freshwater fish culture and transportation, and residue in freshwater fish pose potential risks to human health. Therefore, a throughput method was developed to detect antibiotic and sedative residues in fish, simultaneously quantifying 68 antibiotics and 9 sedatives in freshwater fish using a modified QuEChERS extraction method and UPLC-MS/MS. Matrix-matched calibrations demonstrated good correlation coefficients (R2 > 0.995), with a recovery range of 66.2–118.5%. The intra-day and inter-day relative standard deviation (RSD) were below 9.7% and 12.8%, respectively. The limits of detection (LOD) and quantification (LOQ) were 0.08–1.46 μg/kg and 0.25–4.86 μg/kg, respectively. 68.8% of analytes had weak matrix effects, and 13.0% had moderate matrix effects. In addition, diazepam and many types of antibiotics were detected in30 freshwater fish. The validation parameters were in agreement with the acceptable criteria of the Codex guidelines. The method was effective in analyzing antibiotic and sedative residues in freshwater fish.

Introduction:Pogostemon cablin (PC) is used in traditional Chinese medicine and food, as it exerts pharmacological effects, such as immune-modulatory, antibacterial, antioxidant, antitumor, and antiviral. Currently, the pharmacokinetics (PK) studies of PC mainly focus on individual components. However, research on these individual components cannot reflect the actual PK characteristics of PC after administration. Therefore, the simultaneous determination of multiple components in rat plasma using UPLC-MS/MS was used for the pharmacokinetic study after oral administration of PC extract in this study, providing reference value for the clinical application of PC.Methods: In the present study, a reliable and sensitive ultra-high performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the simultaneous determination of 15 prototype components (vanillic acid, vitexin, verbascoside, isoacteoside, hyperoside, cosmosiin, apigenin, β-rhamnocitrin, acacetin, ombuin, pogostone, pachypodol, vicenin-2, retusin, and diosmetin-7-O-β-D-glucopyranoside) in rat plasma after oral administration of the PC extract. Plasma samples were prepared via protein precipitation using acetonitrile, and icariin was used as the internal standard (IS).Results: The intra-day and inter-day accuracies ranged from −12.0 to 14.3%, and the precision of the analytes was less than 11.3%. The extraction recovery rate of the analytes ranged from 70.6−104.5%, and the matrix effects ranged from 67.4−104.8%. Stability studies proved that the analytes were stable under the tested conditions, with a relative standard deviation lower than 14.1%.Conclusion: The developed method can be applied to evaluate the PK of 15 prototype components in PC extracts of rats after oral administration using UPLC-MS/MS, providing valuable information for the development and clinical safe, effective, and rational use of PC.

Objective： A new method was developed for the simultaneous determination of three kinds of ochratoxins in coffee by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) combined with QuEChERS pretreatment. Methods： Samples were extracted by acetonitri-water-formic acid solution (55∶40∶5) with ultrasonic, salted out with a QuEChERS salt pocket, and then purified with a (ZanChERS-Myco17) column. The chromatographic separation was performed on an ACQUITY UPLC BEH C18 (1.7 μm, 2.1 mm×100 mm) column by gradient elution using a mobile phase comprising 0.2% formic acid aqueous solution and acetonitrile. In the positive ion mode of electric spray, the samples detected were used by multiple reaction monitoring (MRM) modes with internal standard method. Results： Three target substances had a good linear relationship in the range of 0.1~20.0 ng/mL, and the correlation coefficient was greater than 0.999 46. The detection limit and quantitative limit were 0.1~0.2 μg/kg and 0.2~0.7 μg/kg, respectively. Average recoveries at three spiked levels in the coffee matrix were 79.0%~103.3% with a relative standard deviation of 1.5%~7.6%. Conclusion： This method has the advantages of simple sample pretreatment, good reproducibility and high efficiency, and is suitable for the high-throughput detection of multiple ochratoxins in different coffee samples.

AbstractContext Derazantinib—an orally bioavailable, ATP competitive, multikinase inhibitor—has strong activity against fibroblast growth factor receptors (FGFR)2, FGFR1, and FGFR3 kinases. It has preliminary antitumor activity in patients with unresectable or metastatic FGFR2 fusion-positive intrahepatic cholangiocarcinoma (iCCA).Objective This experiment validates a novel sensitive and rapid method for the determination of derazantinib concentration in rat plasma by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS), and applies it to the study of drug-drug interaction between derazantinib and naringin in vivo.Materials and methods A Xevo TQ-S triple quadrupole tandem mass spectrometer was used for mass spectrometry monitoring in selective reaction monitoring (SRM) mode with transitions of m/z 468 96 → 382.00 for derazantinib and m/z 488.01 → 400.98 for pemigatinib, respectively. The pharmacokinetics of derazantinib (30 mg/kg) was investigated in Sprague-Dawley (SD) rats divided into two groups (with the oral pretreatment of 50 mg/kg naringin or not).Results The newly optimized UPLC-MS/MS method was suitable for the determination of derazantinib in rat plasma. It was also successfully employed to evaluate the effect of naringin on derazantinib metabolism in rats. After pretreatment with naringin, there was no significant difference in the pharmacokinetic parameters (AUC0→t, AUC0→∞, t1/2, CLz/F, and Cmax) of derazantinib when compared with derazantinib alone.Conclusion Co-administration of naringin with derazantinib was not associated with significant changes in pharmacokinetic parameters. Thus, this study suggests that the combination of derazantinib with naringin can safely be administered concomitantly without dose adjustment.

AbstractContext Poziotinib and vonoprazan are two drugs mainly metabolized by CYP3A4. However, the drug-drug interaction between them is unknown.Objective To study the interaction mechanism and pharmacokinetics of poziotinib on vonoprazan.Materials and methods In vitro experiments were performed with rat liver microsomes (RLMs) and the contents of vonoprazan and its metabolite were then determined with UPLC-MS/MS after incubation of RLMs with vonoprazan and gradient concentrations of poziotinib. For the in vivo experiment, rats in the poziotinib treated group were given 5 mg/kg poziotinib by gavage once daily for 7 days, and the control group was only given 0.5% CMC-Na. On Day 8, tail venous blood was collected at different time points after the gavage administration of 10 mg/kg vonoprazan, and used for the quantification of vonoprazan and its metabolite. DAS and SPSS software were used for the pharmacokinetic and statistical analyses.Results In vitro experimental data indicated that poziotinib inhibited the metabolism of vonoprazan (IC50 = 10.6 μM) in a mixed model of noncompetitive and uncompetitive inhibition. The inhibitory constant Ki was 0.574 μM and the binding constant αKi was 2.77 μM. In vivo experiments revealed that the AUC(0-T) (15.05 vs. 90.95 μg/mL·h) and AUC(0-∞) (15.05 vs. 91.99 μg/mL·h) of vonoprazan increased significantly with poziotinib pretreatment. The MRT(0-∞) of vonoprazan increased from 2.29 to 5.51 h, while the CLz/F value decreased from 162.67 to 25.84 L/kg·h after pretreatment with poziotinib.Conclusions Poziotinib could significantly inhibit the metabolism of vonoprazan and more care may be taken when co-administered in the clinic.

Yuxiang Xu, Xiaonan Gao, Yunfang Zhu, Qi Zhang, Hongxin Qie, Haopeng Zhao, Jinglin Gao, Mingxia Wang Department of Clinical Pharmacology, The Fourth Hospital of Hebei Medical University, Shijiazhuang, 050011, People’s Republic of ChinaCorrespondence: Mingxia Wang, Department of Clinical Pharmacology, The Fourth Hospital of Hebei Medical University, 12 Jiankang Road, Shijiazhuang, 050011, People’s Republic of China, Tel +86-311-66696233, Email mxia_wang@163.comPurpose: To develop an UPLC-MS/MS method for the quantitative analysis of pentoxifylline in beagle dog plasma and apply it to a pharmacokinetic study of food effect.Methods: Sample separation was achieved using a Kinetex Phenyl-Hexyl column (50 × 2.1 mm, 1.7 μm) with a gradient elution program in 5.5 min after a simple protein precipitation with methanol. Using the mobile phase that made up by 0.2% formic acid and 5mM ammonium formate water (A) and methanol (B). Quantitation was carried out using the positive ionization mode with multiple reaction monitoring (MRM). A randomized, single-dose, two-period crossover study was conducted in six fasted or fed beagles that received 400 mg pentoxifylline sustained-release tablets (Brand name: Shuanling™, CSPC Pharmaceutical Group). WinNonlin® software was used to calculate pharmacokinetic parameters.Results: The linear calibration range was 2– 1000 ng/mL (r2> 0.99). Both intra- and inter-batch precision were less than 6.27%, and the accuracy ranged from 88.65% to 97.18%. Pentoxifylline was readily absorbed in fasted and fed dogs administered a dose of 400 mg (tmax:1.54h vs 1.83h). Compared to the fasted group, the AUC0→t and Cmax in the fed group increased by 1.71-fold and 1.30-fold, respectively. In the fasted group, the AUC0→t and Cmax values were 4684.08 h•ng/mL and 2402.33 ng/mL, respectively. In the fed group, these values were 8027.75 h•ng/mL and 3119.67 ng/mL. The difference in AUC0-t between the fed and fasted group was statistically significant.Conclusion: The novel optimized UPLC-MS/MS assay is an effective tool for the determination of pentoxifylline and has been successfully applied in pharmacokinetic studies of pentoxifylline in beagle dogs. The administration of pentoxifylline sustained-release tablets with food significantly increased the area under the time curve, and it is recommended that they should be administered during or shortly after feeding.Keywords: pentoxifylline, sustained-release tablets, food effect, UPLC-MS/MS, pharmacokinetics

A quick, easy, cheap, effective, rugged and safe pre-treatment method (QuEChERS) was established to evaluate the risk of tetracycline residues in the muscle of Channa argus. The Box-Behnken design (BBD) response surface optimization method was used to optimize the extraction and purification conditions. The samples were extracted with 10 mL acetonitrile (containing 1% formic acid) and 2 mL 0.1 mol/L Na2EDTA-Mcllvaine buffer solution, and the extracts were purified by 150 mg primary secondary amine（PSA）and 200 mg C18, and detected with Ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The result showed that the calibration curve was linear in the range of 0.50~100 ng/mL, with high correlation coefficients (r≥0.999). The limits of detection (LOD) and quantification (LOQ) were 0.25～0.75 μg/kg and 0.84～2.00 μg/kg, respectively. The average recoveries of tetracycline, oxytetracycline, chlortetracycline and doxycycline at three different spiked levels in the muscle of Channa argus were 89.5%～118%, with relative standard deviations (RSDs) of 1.11%～8.88%. This method is simple, sensitive and accurate, and can be used for the rapid quantitative determination of tetracyclines in Channa argus.

Objective: To analyze the differences of chemical composition of Pericarpium Citri Reticulatae 'Chachi' under different aging years, and to provide the theoretical and scientific basis for the aging mechanism of Pericarpium Citri Reticulatae 'Chachi'. Methods: UPLC-MS/MS widely-targeted metabolomics technology was used to detect and analyze the chemical constituents of three kinds of Pericarpium Citri Reticulatae 'Chachi' aged seven years, nine years and eleven years. PCA and OPLS-DA statistical analysis methods were used to identify differential metabolites. Results: A total of 778 compounds of 27 categories were identified. Main components were flavonoids (121) and terpenoids (90). According to the VIP value greater than 1 and the P-value less than 0.05, 121 differential metabolites were screened, 65 differential metabolites of 22 categories between the 11-year aging group and 9-year aging group, 62 differential metabolites of 20 categories between the 11-year aging group and 7-year aging group, 32 differential metabolites of 14 categories between the 9-year aging group and 7-year aging group. The numbers and types of differential metabolites in Pericarpium Citri Reticulatae 'Chachi' tended to increase after aging for 7~11 years. There were differences in metabolites of Pericarpium Citri Reticulatae 'Chachi' under different aging years. With the increase of aging time, the aging time had great influence on flavonoids, terpenoids, amino acid and derivatives. The key pathways with the highest correlation with differential metabolites were: Monoterpenoid biosynthesis, flavone and flavonol biosynthesis, the metabolic pathways involved in amino acid and derivatives (alanine, aspartate and glutamate metabolism, glycine, serine and threonine metabolism, cysteine and methionine metabolism). Conclusion: Widely-targeted metabolomics technology can be used to identify and analyze the chemical components of Pericarpium Citri Reticulatae 'Chachi' with high efficiency. It has established important technical foundation for researching the aging mechanism of Pericarpium Citri Reticulatae 'Chachi'.

Zaiwen Fan,1 Xiaonan Gao,2 Mingxia Wang,2 Ziqiang Tian1 1Department of Thoracic Surgery, The Fourth Hospital of Hebei Medical University, Shijiazhuang, People’s Republic of China; 2Department of Clinical Pharmacology, The Fourth Hospital of Hebei Medical University, Shijiazhuang, People’s Republic of ChinaCorrespondence: Ziqiang Tian, Department of Thoracic Surgery, The Fourth Hospital of Hebei Medical University, 12 Jiankang Road, Shijiazhuang, Hebei, 050011, People’s Republic of China, Email dr_tianzq@126.com Mingxia Wang, Department of Clinical Pharmacology, The Fourth Hospital of Hebei Medical University, 12 Jiankang Road, Shijiazhuang, 050011, People’s Republic of China, Email mxia_wang@163.comAim: The goal of our study was to investigate the effects of single-dose simvastatin and itraconazole application on the pharmacokinetics of erlotinib in rats.Methods: Twenty-one male Sprague-Dawley rats were randomly divided into 3 groups, including erlotinib combined with simvastatin, erlotinib combined with itraconazole and erlotinib alone groups. The rats were given a single dose of 2 mg/kg simvastatin, 15 mg/kg itraconazole or 0.5% sodium carboxymethyl cellulose followed by 12 mg/kg erlotinib. The concentration of erlotinib in rat plasma was determined by UPLC-MS/MS. As internal standard, tinidazole was used for chromatographic analysis on the Kinetex C18 column (100× 2.1 mm, 2.6 μm).Results: Erlotinib was validated in the calibration range of 5– 1000 ng/mL. The lower limit of quantification (LLOQ) was 5 ng/mL. The inter- and intra-day precisions for erlotinib were less than 10.56%, and the accuracies were in the range of 98.61– 104.99%. The validated UPLC-MS/MS method was successfully applied to this study. Compared with the erlotinib alone group, the values of AUC0-t, AUC0-∞, Cmax, Vz/F and t1/2 in the simvastatin group showed no statistical differences among pharmacokinetic parameters (P> 0.05). However, the values of AUC0-t, AUC0-∞ and Cmax, in the itraconazole group were approximately 1.32-fold, 1.32-fold and 1.34-fold higher, and the CL/F was lower than those in the erlotinib alone group; the difference was statistically significant (P< 0.05).Conclusion: Simvastatin had no significant effect on the pharmacokinetics of erlotinib, whereas co-administration of itraconazole considerably increased the exposure of erlotinib. Therefore, we should pay more attention to the potential drug-drug interaction to ensure safety in cancer patient treatment.Keywords: erlotinib, simvastatin, itraconazole, drug–drug interaction, UPLC-MS/MS, pharmacokinetic

Objective: In order to explore the uric acid-lowering activity and mechanism of four traditional Chinese medicines, celery seed, dandelion, chicory and corn silk, and analysis of the main functional components with uric acid-lowering effect in these four kinds of traditional Chinese medicine compound extracts. Methods: Hyperuricemia model in zebrafish was induced by potassium oxazinate and sodium xanthine, then they were randomly divided into control group, model group, allopurinol group (136 µg/mL) and compound extract low, medium, high doses group (250, 500 and 1000 µg/mL), with 30 rats in each group. After continuous intervention for 24 h, the fluorescence level of uric acid and the gene expression levels of hypoxanthine phosphoribosyltransferase1 (HPRT1), glucose transporter9 (GLUT9), and organic anion transporter (OAT1) in zebrafish were measured. Using liquid chromatography-series high resolution mass spectrometer as the main method, and acetonitrile formate water as the mobile phase, gradient elution, ESI ion source positive and negative ion scanning mode, and then combined with software database search and related literature for component identification. Results: Compared with the model group, the low, medium and high dose groups of the compound extract could reduce the blood uric acid level of the hyperuric acid zebrafish model (P

Phellodendron amurense Rupr. is a precious medicinal tree species in northeast China. However, P. amurense resources have been severely destroyed due to uncontrolled overharvest and the limited innovation of new germplasms by traditional cross-breeding. In this study, polyploid breeding was introduced to the improvement program of P. amurense. Fifty-four tetraploid plants of P. amurense were first produced by colchicine-induced adventitious bud chromosome doubling in stem segment explants. The induction frequency reached 36.16% (1.0 g L−1 colchicine solution for 48 h treatment) and 50.00% (2.0 g L−1 colchicine solution for 24 h treatment), respectively, showing the high efficiency of the somatic chromosome doubling based on the organogenesis system. Tetraploidization resulted in significant phenotypic variation, such as larger and thicker leaves, thicker stems, and bigger stomata. Ultra-performance liquid chromatography–mass spectrometry (UPLC–MS/MS) analysis identified 59 differentially accumulated alkaloids (DAAs) between the leaf and stem samples of tetraploids, including 32 upregulated and 27 downregulated in stems. For both leaf and stem samples, 18 DAAs were identified between diploids and tetraploids, with 16 DAAs upregulated in tetraploid leaves and 8 upregulated in tetraploid stems, suggesting that polyploidization caused significant alterations in alkaloid contents in leaves and stems of P. amurense. The contents of the main medicinal compounds, such as berberine, jatrorrhizine, phellodendrine, and palmatine, increased significantly in the leaf and/or stem samples after polyploidization. This finding implied that polyploid breeding might be an effective approach for improving P. amurense, beneficial to preserving and exploiting natural resources.

Despite a higher safety profile compared to vitamin K antagonists, rivaroxaban therapy is still connected with multiple adverse effects, such as a high risk of bleeding. Thus, therapeutic drug monitoring (TDM) of rivaroxaban concentrations is suggested. An alternative to plasma samples can be dried blood spots (DBS), which minimize the cost of sample storage and transport. In this study, we developed a UPLC-MS/MS method for the analysis of rivaroxaban in DBS and plasma samples. Chromatographic separation was achieved on a Zorbax Eclipse Plus C18 column (2.1 × 100 mm; 3.5 µm, Agilent Technologies Inc., Santa Clara, CA, USA) with a mobile phase consisting of water and acetonitrile, both containing 0.1% formic acid. The analytes were detected using a positive ionization mode by multiple reaction monitoring. We validated the method according to ICH guidelines. The precision and accuracy were satisfactory. Extraction recovery was approximately 57% and 66% for DBS and plasma samples, respectively. A high correlation between rivaroxaban concentrations in plasma and DBS samples collected from patients was confirmed with Deming regression. The suitability of both sampling techniques for the rivaroxaban TDM was also verified by Bland–Altman plots based on DBS-predicted and observed plasma concentrations. In addition, we found a significant relationship between rivaroxaban concentrations and coagulation parameters, including prothrombin time (PT) and international normalized ratio (INR).

The aim of individuals consuming health supplements is to attain a robust state through nutritional regulation. However, some unscrupulous manufacturers, motivated by profit, fraudulently incorporate drugs or unauthorized components with therapeutic effects into the product for instant product performance enhancement. The long-term use of these products may inadvertently inflict harm on human health and fail to promote nutritive healthcare. The illegal inclusion of these substances is prevalent in kidney-tonifying and sexuality-enhancing products. Developing effective analytical methods to identify these products and screen for illegal added ingredients can effectively prevent such products from reaching and remaining on the market. A target screening method for the detection and quantification of 90 phosphodiesterase type 5 inhibitors (PDE-5is) in 5 kinds of health products was developed and validated. The type of dietary supplements varied from tablets, capsules, and protein powder to wine and beverages. Sample preparation was completed with a one-step liquid phase extraction. The screening process of 90 PDE-5is was done efficiently within 25 min by ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) using the dynamic multiple reaction monitoring (dMRM) technique. The LODs of 90 PDE-5is were detected at levels ranging from 25 to 85 ng/g or ng/mL. This novel targeting methodology was effective and can be applied to routine market supervision. Among 286 batches of samples, 8 batches were found to be positive. Three kinds of PDE-5is were first detected in healthy products. The screening method demonstrated herein will be a promising and powerful tool for rapid screening of PDE-5is.

Our previously reported, first validated, UPLC-MS/MS-based simultaneous analysis of five human milk B-vitamins revealed severe matrix effects. High levels of endogenous lactose fouled the electrospray ionization source affecting the analysis. We evaluated solid-phase extraction (SPE), liquid-solid extraction (LSE), protein precipitation (PPT), and liquid chromatography effluent diversion for lactose-removal. SPE failed to separate lactose from vitamins; LSE using 2-propanol reduced lactose and vitamin recoveries. PPT-solvent, milk volume, and reconstitution solvent influenced flavin adenine dinucleotide, pyridoxal and nicotinamide recoveries. Using an optimized LC-gradient enabled chromatographic separation of lactose from vitamins and its removal using a post-column switch-valve. Only 40 µL milk was subjected to methanol-PPT and non-polar matrix removal by methyl tert-butyl ether. B-vitamin recoveries were established (81.9-118.6%; CV ≤ 11.9%; precision: 4.9-13.7%) with greatly reduced matrix effects, and improved process efficiency, and recovery.

An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed for the simultaneous quantitation of doxorubicin (DOX) and sorafenib (SOR) in rat plasma. Chromatographic separation was performed using a reversed-phase column C18 (1.7 μm, 1.0 × 100 mm Acquity UPLC BEH™). The gradient mobile phase system consisted of water containing 0.1% acetic acid (mobile phase A) and methanol (mobile phase B) with a flow rate of 0.40 mL/min over 8 min. Erlotinib (ERL) was used as an internal standard (IS). The quantitation of conversion of [M + H]+, which was the protonated precursor ion, to the corresponding product ions was performed using multiple reaction monitoring (MRM) with a mass-to-charge ratio (m/z) of 544 > 397.005 for DOX, 465.05 > 252.03 for SOR, and 394 > 278 for the IS. Different parameters were used to validate the method including accuracy, precision, linearity, and stability. The developed UPLC–MS/MS method was linear over the concentration ranges of 9–2000 ng/mL and 7–2000 ng/mL with LLOQ of 9 and 7 ng/mL for DOX and SOR, respectively. The intra-day and inter-day accuracy, expressed as % relative standard deviation (RSD%), was below 10% for both DOX and SOR in all QC samples that have drug concentrations above the LLOQ. The intra-day and inter-day precision, expressed as percent relative error (Er %), was within the limit of 15.0% for all concentrations above LLOQ. Four groups of Wistar rats (250–280 g) were used to conduct the pharmacokinetic study. Group I received a single intraperitoneal (IP) injection of DOX (5 mg/kg); Group II received a single oral dose of SOR (40 mg/kg), Group III received a combination of both drugs; and Group IV received sterile water for injection IP and 0.9% w/v sodium chloride solution orally to serve as a control. Non-compartmental analysis was used to calculate the different pharmacokinetic parameters. Data revealed that coadministration of DOX and SOR altered some of the pharmacokinetic parameters of both agents and resulted in an increase in the Cmax and AUC and reduction in the apparent clearance (CL/F). In conclusion, our newly developed method is sensitive, specific, and can reliably be used to simultaneously determine DOX and SOR concentrations in rat plasma. Moreover, the results of the pharmacokinetic study suggest that coadministration of DOX and SOR might cause an increase in exposure of both drugs.

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.

A reliable and sensitive UPLC-MS/MS method coupled with HLB-SPE was developed for simultaneous determination of T-2 and its modified forms (HT-2, NEO, T-2-triol, T-2-tetraol, T-2-3G, and HT-2-3G) in cereals and cereal-based products. Acceptable linearity (R2 ≥ 0.99), limits of quantitation (0.5–10.0 μg/kg), intra-day precision (RSD