Abstract In this study, we firstly established and verified a method by ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) for the analysis of vilazodone and its metabolite M10 in rat plasma, then this method was used to explore the pharmacokinetics of vilazodone and M10 present or absence of 80 mg/kg bergenin in rats. Protein precipitation with acetonitrile was used to prepare the samples in this research. The mobile phase for liquid chromatography was consisted of 0.1% formic acid aqueous solution and acetonitrile. Brexpiprazole was used as the internal standard (IS), and the multiple reaction monitoring (MRM) mode was used for detection. The verification items required by the US Food and Drug Administration (FDA) guidelines such as selectivity, sensitivity, linearity, stability, recovery and matrix effect of this method were all met the standards. Besides, rats were used to explore the drug-drug interaction between vilazodone and bergenin, which were divided into two groups, and separately gavaged with the same-volume of carboxymethyl cellulose sodium (CMC-Na) solution and 80 mg/kg bergenin, respectively. The results showed that bergenin significantly affected the metabolism of vilazodone. It suggested that there was a potential drug-drug interaction between bergenin and vilazodone in rats. In clinical application, we should pay attention to the dose of vilazodone when in combination with bergenin.
Background: Vericiguat, as a new stimulator of soluble guanylate cyclase (sGC), was recently approved as a first-in-class treatment for reducing risks in patients with ejection fraction less than 45 percent and heart failure (HF) in the USA. Objective: The main aim of the present experiment was to establish an acceptable, sensitive assay based on ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) for quantitatively analyzing the plasma concentration levels of vericiguat in rats, and to further evaluate the effect of apigenin on the metabolism of vericiguat in vivo. Methods: In sample processes, acetonitrile was finally chosen for quickly precipitating protein. The levels of vericiguat in plasma were analyzed by a Xevo TQ-S triple quadrupole tandem mass spectrometry (Milford, MA, USA) in a positive ion mode. Results: The scope of the calibration standard for vericiguat ranged from 0.5 to 1000 ng/mL, where a great linearity was acceptable. The lower limit of quantification (also called LLOQ) of vericiguat presented the sensitivity of this assay was evaluated as low as 0.5 ng/mL. Additionally, selectivity, accuracy and precision, extraction recovery, matrix effect, and stability were all verified. Subsequently, this approach also supported to assess the plasmatic concentrations of vericiguat from an interaction survey on herb-- drug, in which oral administration of apigenin (20 mg/kg) obviously increased the plasmatic levels of vericiguat and altered the pharmacokinetics of vericiguat in rats. Conclusion: These results would help us to further understand the pharmacokinetic properties of vericiguat when co-administration with apigenin, and to avoid unexpected clinical risks in the future.
Elagolix, as a competitive gonadotropin-releasing hormone (GnRH) receptor antagonist, has been recently approved by the US FDA for the management of moderate to severe pain due to endometriosis in women. In this study, we developed and verified an analysis assay to detect the concentration level of elagolix in plasma from rats after sample preparation based on a newly validated ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) technique in this study. The process of sample preparation used acetonitrile for a quick and easy protein precipitation method and diazepam was engaged as the internal standard (IS). Then, gradient elution was used to elute elagolix and IS. The mobile phase used in the present experiment was consisted of solvent A (acetonitrile) and solvent B (water having formic acid with the volume ratio of 0.1%), and the type of the C18 column used was named Acquity UPLC BEH C18 column with the specification of 2.1 mm × 100 mm, 1.7 μm. Multiple reaction monitoring (MRM) in positive ion mode for the experiment was engaged to detect the level of elagolix with electrospray ionization (ESI) source by m/z 632.4 → 529.5 transition for quantification and m/z 632.4 → 177.1 transition for qualification. It was found that the method in the scope of 1–2000 ng/mL indicated excellent linearity (r2 > 0.9983). The precision of this assay for intra-day was between 3.5 and 5.5%, and for inter-day was between 9.4 and 12.7%, respectively; the accuracy was 1.2–13.9% for the intra- and inter-day. The stability, extraction recovery, and matrix effect of the method were all in accordance with the rules of assay validation in biological medium proposed by FDA, whose application was also successfully used to determine the concentration of plasma elagolix from an experiment on pharmacokinetic investigation after oral administration of 15 mg/kg elagolix.
Vortioxetine, as a new therapeutic drug of major depressive disorder (MDD), was approved for MDD in the USA. The significance of our study in this paper was to explore the inhibitory effect of propafenone on the metabolism of vortioxetine through the in vivo and in vitro experiments. In the vitro experiments, we added a series of concentrations of propafenone into an incubation system as the inhibitors and calculated the half-maximal inhibitory concentration (IC50) of propafenone on vortioxetine metabolism in human liver microsomes (HLMs) and rat liver microsomes (RLMs). Twelve male Sprague-Dawley (SD) rats were included in the vivo experiments. We randomly divided them into Group A (control group) and Group B (90 mg/kg propafenone). 30 min later, a single oral dose of 4.0 mg/kg vortioxetine was administrated to each rat. Then, we used ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) to determine the concentrations of vortioxetine and its metabolite Lu AA34443. From our results, it indicated that propafenone inhibited the metabolic rate of vortioxetine in the vitro studies with the IC50 of 0.48 μM and 16.5 μM for HLMs and RLMs, respectively. And, propafenone could competitively inhibit vortioxetine in both HLMs and RLMs for the inhibitory mechanisms. Moreover, a single oral dose of 90 mg/kg propafenone obviously enchanced the exposure of vortioxetine in rats, but not Lu AA34443. Combined with the vitro and vivo data, propafenone showed the inhibitory effect on vortioxetine metabolism. Thus, more attention to the safety of vortioxetine in clinic should be paid when taking it with propafenone in combination for the therapy.
Infigratinib (INF) is a fibroblast growth factor receptor (FGFR)-specific tyrosine kinase inhibitor for the therapy of advanced cholangiocarcinoma. However, CYP3A4 polymorphisms and CYP3A4 inducers or inhibitors might affect the pharmacokinetics of INF. Clinical evaluation of drug-drug interactions and adverse effects in these patients was necessary with reference to INF levels in vivo. The presently conducted study optimized a reproducible and rapid ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) analytic technique, which was validated and applied to determine INF concentrations in Sprague-Dawley (SD) rat plasma and pharmacokinetic studies. Protein was precipitated by adding acetonitrile to plasma samples, followed by gradient elution on a Waters Acquity UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) for complete chromatographic separation of the analyte and derazantinib (used as internal standard, IS). A gradient elution of 0.1% formic acid aqueous solution and acetonitrile at a flow rate of 0.30 mL/min was applied as the mobile phase for this analysis. The ion transitions of INF and IS were m/z 599.88 → 313.10 and m/z 468.96 → 382.00 during UPLC-MS/MS detection, respectively, by multiple reaction monitoring (MRM). The methodological validation demonstrated that INF presented high linearity over the concentration of 2–600 ng/mL. The lower limit of quantification (LLOQ) for this experiment was 2 ng/mL, of which the precisions and accuracies were within the permissible levels. Inter-day and intra-day precisions were demonstrated to be within reasonable limits, which were within 15%, and the accuracies were determined to be between 2.2% and 11.4%. Moreover, the values of recovery, stability and matrix effect of INF were within the limits of acceptability. The pharmacokinetics of INF in SD rats was investigated by gavage administration of 10 mg/kg INF, followed by the application of the developed UPLC-MS/MS analytical method to detect the content in plasma and derive the main pharmacokinetic parameters.
We established a method based on ultra performance liquid chromatography tandem mass spectrometry (UPLC‒MS/MS) to quantitatively measure tepotinib, which was validated as acceptable and used in the evaluation of food-drug interactions between tepotinib and naringenin in rats. We used pemigatinib as the internal standard (IS), and acetonitrile and 0.1% formic acid aqueous solution constituted the mobile phase. To extract the target analyte, acetonitrile was used for protein precipitation (PPT). For UPLC‒MS/MS, we performed liquid chromatography using a C18 column, and mass spectrometry was performed in positive multiple reaction monitoring (MRM) mode. Excellent linearity was shown in the range of 0.1–500 ng/mL, and the coefficient of correlation was > 0.99. Notably, the lower limit of quantification (LLOQ) for tepotinib was determined to be 0.1 ng/mL. The intra- and inter-day accuracy of tepotinib ranged from − 1.7 to 7.3%, while the precision was ≤ 8.4%, at three concentrations except LLOQ. The recovery of each substance was ≥ 81.2%, and the matrix effects were within 90.5-98.6%. The stabilities of all analytes under different conditions met all requirements for quantitation in plasma samples. The relevant parameters, such as LLOQ, were evaluated in accordance with the principles of the Food and Drug Administration (FDA) biological verification method. Food-drug interaction study had shown that the plasma concentration of tepotinib could be significantly increased, accompanied by a decrease in clearance rate when administered with 50 mg/kg naringenin. The results showed that naringenin could increase the plasma concentration and decrease the clearance rate of tepotinib when naringenin and tepotinib were administered at the same time.
This study aimed to explore the effect of baicalein on the pharmacokinetics of cilostazol (CLZ) and its two metabolites 3,4-dehydro cilostazol (3,4-CLZ) and 4'-trans-hydroxy cilostazol (4'-CLZ) in rats using a newly established ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method. Ticagrelor was used as an internal standard (IS), then cilostazol and its two metabolites were separated by means of a UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) using gradient elution method with 0.4 ml/min of flow rate. Acetonitrile as organic phase and water with 0.1% formic acid as aqueous phase constructed the mobile phase. Selective reaction monitoring (SRM) mode and positive ion mode were preferentially chosen to detect the analytes. Twelve SD rats were divided into two groups (n = 6) when CLZ was administered orally (10 mg/kg) with or without oral baicalein (80 mg/kg). The selectivity, linearity, recovery, accuracy, precision, matrix effect and stability of UPLC-MS/MS assay were satisfied with the standards of United States Food and Drug Administration guidelines. In control group, AUC0-∞ and Cmax of CLZ were 2,169.5 ± 363.1 ng/ml*h and 258.9 ± 82.6 ng/ml, respectively. The corresponding results were 3,767.6 ± 1,049.8 ng/ml*h and 308.6 ± 87.9 ng/ml for 3, 4-CLZ, 728.8 ± 189.9 ng/ml*h and 100.3 ± 51.3 ng/ml for 4'-CLZ, respectively. After combination with baicalein, AUC0-∞ and Cmax of CLZ were 1.48, 1.38 times higher than the controls. Additionally, AUC0-∞ and Cmax were separately decreased by 36.12 and 19.54% for 3,4-CLZ, 13.11 and 44.37% for 4'-CLZ. Baicalein obviously alters the pharmacokinetic parameters of CLZ, 3,4-CLZ and 4'-CLZ in rats. These results suggested that there was a potential drug-drug interaction between baicalein and CLZ. Therefore, it must raise the awareness when concomitant use of CLZ with baicalein, the dosage regimen of CLZ should be taken into consideration, if this result is confirmed in clinical studies.
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