Characterization of Alpelisib in Rat Plasma by a Newly Developed UPLC-MS/MS Method: Application to a Drug-Drug Interaction Study

Alpelisib, an oral selective and small-molecule phosphoinositide 3-kinase inhibitor, was lately approved in the United States to treat breast cancer. A sensitive method to quantify alpelisib levels in rat plasma on the basis of ultra-performance liquid chromatography–tandem mass spectrometry technique was established and validated, which was successfully employed to explore the effects of CYP3A4 inhibitors on alpelisib pharmacokinetics in rats. A C18 column named Acquity UPLC BEH C18 was applied to achieve the separation of alpelisib and internal standard duvelisib after protein precipitation with acetonitrile. The mobile phase in this study had two components, namely, acetonitrile and water having 0.1% formic acid, and a program with gradient elution method was used at a flow rate of 0.40 ml/min. Mass spectrometry in a positive multiple reaction monitoring mode was operated. In the scope of 1–5,000 ng/ml, this assay had excellent linearity. Our newly developed assay was verified in all aspects of bioanalytical method validation, involving lower limit of quantification, selectivity, accuracy and precision, calibration curve, extraction recovery, matrix effect, and stability. Then, this assay was used to detect the plasma levels of alpelisib from a drug-drug interaction investigation, where ketoconazole remarkably increased the plasma concentration of alpelisib and changed alpelisib pharmacokinetics more than itraconazole. This study will help better understand the pharmacokinetic properties of alpelisib, and further clinical studies should be done to confirm this result in patients.

Download Full-text

Validation of an UHPLC–MS/MS Method for the Determination of Glaucocalyxin A, a Novel Potent Negative Akt Regulator in Rat Plasma, Lung and Brain Tissues: Application to a Pharmacokinetic Study

Journal of Chromatographic Science ◽

10.1093/chromsci/bmz095 ◽

2019 ◽

Vol 58 (3) ◽

pp. 234-240

Author(s):

Zhipeng Deng ◽

Qian Liu ◽

Jixiang He ◽

Sixi Zhang ◽

Wei Zhou

Keyword(s):

Mass Spectrometry ◽

Intravenous Administration ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Rat Plasma ◽

Mass Spectrometry Analysis ◽

Active Constituent ◽

Limit Of Quantification ◽

Relative Standard ◽

Brain Tissues

Abstract Glaucocalyxin A, a novel potent negative Akt regulator, is a major active constituent of Rabdosia japonica. A simple, specific and sensitive ultra-high pressure liquid chromatography tandem mass spectrometry (UHPLC–MS/MS) method was developed and validated for the quantification of glaucocalyxin A in rat plasma, lung and brain tissues after intravenous administration. Sample preparation was carried out through a simple liqiud–liquid extraction with ethyl acetate using bullatine A as internal standard. The chromatographic separation was achieved by using an Agilent ZORBAX Eclipse Plus C18 column (3.0 mm × 100 mm, 1.8 μm) with a mobile phase of acetonitrile and water containing 0.1% formic acid in a gradient elution. Mass spectrometry analysis was conducted in positive ionization mode with multiple reaction monitoring transitions at m/z 333.2 → 157.1 for glaucocalyxin A and m/z 344.2 → 128.1 for IS. Calibration curves were linear over the ranges of 20.0–4000 ng/mL for both plasma and tissue samples (r2 > 0.99). The Lower limit of quantification (LLOQ) was 0.284 ng/mL. The intra-day and inter-day precision relative standard deviation (RSD%) were <14.9%, while the accuracy ranged from −12.5 to 17.0% for LLOQ and quality control samples. This UHPLC–MS/MS method was successfully applied in the pharmacokinetics, lung and brain tissue distributions of glaucocalyxin A after intravenous administration.

Download Full-text

Determination of sarecycline by UPLC-MS/MS and its application to pharmacokinetic study in rats

Acta Chromatographica ◽

10.1556/1326.2020.00796 ◽

2020 ◽

Author(s):

Yonghui Shen ◽

Deru Meng ◽

Feifei Chen ◽

Hui Jiang ◽

Liming Hu ◽

...

Keyword(s):

Pharmacokinetic Study ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Gradient Elution ◽

Rat Plasma ◽

Chronic Inflammatory Disease ◽

Linear Calibration ◽

Limit Of Quantification ◽

Acquity Uplc ◽

Intravenous And Oral Administration

AbstractSarecycline is a narrow-spectrum antibiotic for the treatment of acne, which is a chronic inflammatory disease of the hair follicle sebaceous glands. In the study, UPLC-MS/MS was used to establish a rapid and accurate analytical method. The sarecycline was determined with poziotinib as internal standard (IS) in rat plasma. An ACQUITY UPLC HSS T3 column (2.1 × 100 mm, 1.8 μm) could performe chromatographic separation with the mobile phase (methanol: water of 0.1% formic acid) with gradient elution. The ions of target fragment were m/z 488.19→410.14 for sarecycline and m/z 492.06→354.55 for poziotinib, which could quantify the electrospray ionization of positive multiple reaction monitoring (MRM) mode. The linear calibration curve of the concentration range was 1–1,000 ng/mL for sarecycline with a lower limit of quantification (LLOQ) of 1 ng/mL. The mean recovery was between 82.46 and 95.85% for sarecycline and poziotinib in rat plasma. RSD for precision of inter-day and intra-day were between 3.24 and 13.36%, and the accuracy ranged from 105.26 to 109.75%. The developed and validated method was perfectly used in the pharmacokinetic study and bioavailability of sarecycline after intravenous and oral administration in rats.

Download Full-text

High-throughput LC-MS/MS Method for Simultaneous Determination of Pantoprazole and Atorvastatin in Rat Plasma: Application to a Pharmacokinetic Interaction Study

Current Drug Metabolism ◽

10.2174/1389200222666210520090632 ◽

2021 ◽

Vol 22 ◽

Author(s):

Jian Le ◽

Yuehua Liao ◽

Shengni Li ◽

Xiujuan Chen ◽

Zhanying Hong

Keyword(s):

Drug Interaction ◽

Multiple Reaction Monitoring ◽

Pharmacokinetic Interaction ◽

Rat Plasma ◽

Interaction Study ◽

Triple Quadrupole Mass Spectrometer ◽

Internal Standards ◽

Spectrometric Data ◽

Drug Drug Interaction

Background: Pantoprazole and atorvastatin are often used jointly in the clinic. The drug-drug interaction of pantoprazole and atorvastatin is worthy of being investigated. Objective: A highly rapid, sensitive, and selective LC-MS/MS method was developed for simultaneous quantification of pantoprazole and atorvastatin in rat plasma. Methods: Omeprazole and atorvastatin-d5 were used as the internal standards (ISs) of pantoprazole and atorvastatin, respectively. Simple protein precipitation was used to extract analytes from 50.0 μL plasma samples. Results: The chromatographic separation was achieved on a C18 column and the total chromatographic run time was 3.2 min. Acquisition of mass spectrometric data was performed on a triple-quadrupole mass spectrometer in multiple- reaction-monitoring (MRM) mode with an ESI source using the transition m/z 384→ 200 for pantoprazole and m/z 559.4→ 440.2 for atorvastatin, respectively. The method was validated over the concentration range of 20.0 ∼ 5000 ng/mL for pantoprazole and 1.00 ∼ 250 ng/mL for atorvastatin. All the validation results, including linearity, specificity, precision, accuracy, extraction recovery, matrix effect, and stability, met the acceptance criteria as per FDA guidelines. Conclusion: This method was successfully applied to a pharmacokinetic interaction study in Wistar rats. The results revealed significant evidence for the drug-drug interaction between pantoprazole and atorvastatin.

Download Full-text

Development and Validation of a Sensitive UHPLC-MS/MS Method for the Measurement of Gardneramine in Rat Plasma and Tissues and its Application to Pharmacokinetics and Tissue Distribution Study

Molecules ◽

10.3390/molecules24213953 ◽

2019 ◽

Vol 24 (21) ◽

pp. 3953 ◽

Cited By ~ 1

Author(s):

Zhao ◽

Tan ◽

Chen ◽

Sun ◽

Wang ◽

...

Keyword(s):

Multiple Reaction Monitoring ◽

Indole Alkaloid ◽

Internal Standard ◽

Gradient Elution ◽

Rat Plasma ◽

Limit Of Quantification ◽

Tissue Samples ◽

Distribution Study

As a novel monoterpenoid indole alkaloid, gardneramine has been confirmed to possess excellent nervous depressive effects. However, there have been no reports about the measurement of gardneramine in vitro and in vivo. The motivation of this study was to establish and validate a specific, sensitive, and robust analytical method based on UHPLC-MS/MS for quantification of gardneramine in rat plasma and various tissues after intravenous administration. The analyte was extracted from plasma and tissue samples by protein precipitation with methanol using theophylline as an internal standard (I.S.). The analytes were separated on an Agilent ZORBAX Eclipse Plus C18 column using a gradient elution of acetonitrile and 0.1% formic acid in water at a flow rate of 0.3 mL/min. Gardneramine and I.S. were detected and quantified using positive electrospray ionization in multiple reaction monitoring (MRM) mode with transitions of m/z 413.1→217.9 for gardneramine and m/z 181.2→124.1 for I.S.. Perfect linearity range was 1–2000 ng/mL with a correlation coefficient (r2) of ≥0.990. The lower limit of quantification (LLOQ) of 1.0 ng/mL was adequate for application to different preclinical studies. The method was successfully applied for determination of gardneramine in bio-samples.

Download Full-text

Development and Validation of UPLC-MS/MS Method for Determination of Enasidenib in Rat Plasma and Its Pharmacokinetic Application

International Journal of Analytical Chemistry ◽

10.1155/2020/5084127 ◽

2020 ◽

Vol 2020 ◽

pp. 1-7

Author(s):

Shuang-long Li ◽

Yong-liang Zhu ◽

Yi Zhang ◽

Shu-han Liu ◽

Xiang-die Wang ◽

...

Keyword(s):

Mass Transfer ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Gradient Elution ◽

Rat Plasma ◽

Bioanalytical Method Validation ◽

Fda Approval ◽

Elution Method ◽

Acquity Uplc ◽

Development And Validation

In our research, a straightforward UPLC-MS/MS method, with diazepam as the internal standard (IS), was proposed and acknowledged to determine the concentrations of enasidenib in rat plasma. When preparing the sample, we used acetonitrile for protein precipitation. The gradient elution method was used, and the mobile phase was acetonitrile and 0.1% formic acid. Diazepam was used as the IS. We used the Acquity UPLC BEH C18 column to separate enasidenib and IS. Under the positive ion electrospray ionization (ESI) source conditions, the mass transfer pairs of enasidenib were monitored by multiple reaction monitoring (MRM) to be m/z 474.2 ⟶ 456.1 and m/z 474.2 ⟶ 267.0, and the IS mass transfer pairs were m/z 285.0 ⟶ 154.0. Enasidenib had good linearity (r2 = 0.9985) in the concentration range of 1.0–1000 ng/mL. Besides, the values of intraday and interday precision were 2.25–8.40% and 3.94–5.46%, respectively, and the range of the accuracy values varied from −1.44 to 2.34%. Matrix effect, extraction recovery, and stability were compliant with FDA approval guidelines in terms of bioanalytical method validation. We had established a new method that had been applied to the pharmacokinetic study of enasidenib in rats.

Download Full-text

Ultraperformance Liquid Chromatography–Tandem Mass Spectrometry Assay for Iohexol in Human Serum

Clinical Chemistry ◽

10.1373/clinchem.2008.121533 ◽

2009 ◽

Vol 55 (6) ◽

pp. 1196-1202 ◽

Cited By ~ 24

Author(s):

Thomas M Annesley ◽

Larry T Clayton

Keyword(s):

Mass Spectrometry ◽

Liquid Chromatography ◽

Human Serum ◽

Matrix Effects ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Structural Analog ◽

Limit Of Quantification ◽

Iodinated Contrast ◽

Liquid Chromatography Tandem Mass

Abstract Background: Iohexol is an iodinated contrast dye that has been shown to be useful in the estimation of glomerular filtration rate (GFR) in patients with suspected renal insufficiency. We developed and validated an ultraperformance liquid chromatography (UPLC)–triple quadrupole mass spectrometry (MS/MS) assay for quantifying iohexol in human serum. Methods: Sample preparation involved dilution of 50 μL serum with 400 μL water, followed by protein precipitation with zinc sulfate and methanol containing the structural analog ioversol as the internal standard. After 1:20 dilution of the supernatant with water, 5 μL was injected into the UPLC-MS/MS system. Chromatography was performed using a Waters Oasis HLB 5-μm particle size, 2.1 × 20 mm column maintained at 50 °C. We used a 1-step acetonitrile/0.1% formic acid gradient to elute the compounds of interest at a common retention time of 0.96 min. The multiple reaction monitoring transitions used for integration and quantification were m/z 821.7→803.7 for iohexol and m/z 807.9→589.0 for ioversol in the electrospray positive ionization mode. Results: The assay was linear from 2.5 mg/L (lower limit of quantification) to 1500 mg/L iohexol, with a mean extraction efficiency of >99%. Recovery of nominal target concentrations was 99%–102%. Interassay imprecision ranged from 7.9% at a concentration of 2.5 mg/L to 4.1% at 1000 mg/L. Ion suppression studies showed no matrix effects on the ionization of the 2 compounds. Conclusions: This rapid UPLC-MS/MS method can be successfully used for quantifying iohexol in human serum. .

Download Full-text

Development of UHPLC-MS/MS Method for Indirubin-3′-Oxime Derivative as a Novel FLT3 Inhibitor and Pharmacokinetic Study in Rats

Molecules ◽

10.3390/molecules25092039 ◽

2020 ◽

Vol 25 (9) ◽

pp. 2039

Author(s):

Na Yoon Kim ◽

Yong-Chul Kim ◽

Yoon Gyoon Kim

Keyword(s):

Multiple Reaction Monitoring ◽

Internal Standard ◽

Rat Plasma ◽

Reaction Monitoring ◽

Spectrometry Method ◽

Limit Of Quantification ◽

Flt3 Inhibitor ◽

Pharmacokinetic Properties ◽

Safety Guidelines ◽

The U.S

This study aimed to develop and validate a sensitive liquid chromatography-coupled tandem mass spectrometry method for the quantification of LDD-2614, an indirubin derivative and novel FLT3 inhibitor, in rat plasma. In addition, the developed analytical method was applied to observe the pharmacokinetic properties of LDD-2614. Chromatographic separation was achieved on a Luna omega C18 column using a mixture of water and acetonitrile, both containing 0.1% formic acid. Quantitation was performed using positive electrospray ionization in a multiple reaction monitoring (MRM) mode. The MRM transitions were optimized as m/z 426.2→113.1 for LDD-2614 and m/z 390.2→113.1 for LDD-2633 (internal standard), and the lower limit of quantification (LLOQ) for LDD-2614 was determined as 0.1 ng/mL. Including the LLOQ, the nine-point calibration curve was linear with a correlation coefficient greater than 0.9991. Inter- and intraday accuracies (RE) ranged from −3.19% to 8.72%, and the precision was within 9.02%. All validation results (accuracy, precision, matrix effect, recovery, stability, and dilution integrity) met the acceptance criteria of the U.S. Food and Drug Administration and the Korea Ministry of Food and Drug Safety guidelines. The proposed method was validated and demonstrated to be suitable for the quantification of LDD-2614 for pharmacokinetics studies.

Download Full-text

Simultaneous determination and pharmacokinetics studies of five isoflavones in rat plasma by UHPLC-MS/MS after oral administration of Radix Astragali extract

Acta Chromatographica ◽

10.1556/1326.2020.00845 ◽

2021 ◽

Author(s):

Jianwei Han ◽

Wenbo Zhu ◽

Ling Yu ◽

Yajun Chen ◽

Gaosong Wu ◽

...

Keyword(s):

Oral Administration ◽

High Performance ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Rat Plasma ◽

Tandem Mass ◽

Radix Astragali ◽

Limit Of Quantification ◽

Liquid Chromatography Tandem Mass ◽

Mass Spectrometery

AbstractA rapid, sensitive and convenient method based on ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) was developed and validated for the simultaneous quantification of calycosin-7-O-β-d-glucoside (CCSG), ononin, calycosin, (6aR,11aR)-9,10-dimethoxypterocarpan-3-O-β-d-glucopyanoside (DPPG), and 7,2′-dihydroxy-3′,4′-dimethoxyisoflavan-7-O-β-d-glucopyanoside (DIFG) in rat plasma after oral administration of the methanol extraction from Radix Astragali. Theophylline played the role of internal standard (IS). Preparation of plasma samples by liquid-liquid extraction method with ethyl acetate after precipitation of protein with methanol. The analytes were detected with a triple quadrupole tandem mass spectrometery (MS) in multiple reaction monitoring (MRM) mode and a positive ion electrospray ionization (ESI). The method was validated with the concentration ranges of 1.96–62.69 ng/mL for CCSG, 1.70–54.5 ng/mL for ononin, 1.85–59.06 ng/mL for calycosin, 2.14–137.24 ng/mL for DPPG and1.96–125.25 ng/mL for DIFG, respectively. The method had the lower limit of quantification (LLOQ) with 0.49, 0.21, 0.92, 1.07, and 0.98 ng/mL for CCSG, ononin, calycosin, DPPG and DIFG respectively, and the precision less than 10%. The RSD of the accuracy was in the range of −4.35–8.91%. The results may be helpful to provide more accurate references to clinical application of this herb.

Download Full-text

Pharmacokinetics of Lusutrombopag, a Novel Thrombopoietin Receptor Agonist, in Rats by UPLC-MS/MS

International Journal of Analytical Chemistry ◽

10.1155/2020/7290470 ◽

2020 ◽

Vol 2020 ◽

pp. 1-6

Author(s):

Bo Wang ◽

Feifei Chen ◽

Quan Zhou ◽

Yunfang Zhou ◽

Deru Meng ◽

...

Keyword(s):

Receptor Agonist ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Gradient Elution ◽

Rat Plasma ◽

Linear Calibration ◽

Limit Of Quantification ◽

Mass Spectrometric Measurement ◽

Thrombopoietin Receptor ◽

Positive Ion

Lusutrombopag is a second oral thrombopoietin (TPO) receptor agonist that selectively acts on human TPO receptors. In the study, UPLC-MS/MS was used to establish a selective and sensitive method to determine lusutrombopag with poziotinib as IS (internal standard) in rat plasma. Samples were prepared by precipitating protein with acetonitrile as a precipitant. Separation of lusutrombopag and poziotinib was performed on a CORTECS UPLC C18 column (2.1 ∗ 50 mm, 1.6 μm). The mobile phase (acetonitrile and water containing 0.1% formic acid) with gradient elution was set at a flow rate of 0.4 ml/min. The mass spectrometric measurement was conducted under positive ion mode using multiple reaction monitoring (MRM) of m/z 592.97 ⟶ 491.02 for lusutrombopag and m/z for poziotinib (IS) 492.06 ⟶ 354.55. The linear calibration curve of the concentration range was 2–2000 ng/ml for lusutrombopag, with a lower limit of quantification (LLOQ) of 2 ng/ml. RSD of interday and intraday precision were both no more than 9.66% with the accuracy ranging from 105.82% to 108.27%. The extraction recovery of lusutrombopag was between 82.15% and 90.34%. The developed and validated method was perfectly used in the pharmacokinetic study of lusutrombopag after oral administration in rats.

Download Full-text

BIO-ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF DECITABINE AND CEDAZURIDINE IN HUMAN PLASMA USING LC-MS/MS

International Journal of Applied Pharmaceutics ◽

10.22159/ijap.2021v13i5.41744 ◽

2021 ◽

pp. 257-262

Author(s):

SAI PRUDHVI N. ◽

VENKATESWARLU B. S. ◽

KUMUDHAVALLI M. V. ◽

MURUGANANTHAM V.

Keyword(s):

Mass Spectrometry ◽

Quality Control ◽

Human Plasma ◽

Method Development ◽

Limit Of Detection ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Simultaneous Estimation ◽

Limit Of Quantification ◽

Validation Parameters

Objective: The present work aimed to develop a novel, reliable and accurate Liquid Chromatography-Mass Spectrometry/Mass spectrometry (LC-MS/MS) method for the simultaneous quantification of Decitabine and Cedazuridine a combined medication used for the treatment of chronic myelomonocytic leukemia in human plasma. Methods: Talazoparib drug is used as an internal standard in the study. Both the analytes and internal standard were isolated from 100 ml plasma samples by liquid-liquid extraction and then chromatographed on Zorbax SB-CN (4.6 mm×75 mm, 3.5 µm) column with a mobile phase consisting of 0.1 % ammonium formate and methanol in the ratio of 65:45 (v/v) pumped at 0.5 ml/min. The method had a chromatographic total run time of 5 min. Results: The developed method gave a symmetric peak at a retention time of 1.7 min for Decitabine, 2.2 min for Cedazuridine, 3.5 min for Talazoparib and satisfied all the peak properties as per USP guidelines. The mass spectral characterization of separated analytes in the LC method was performed using a mass detector operated at Multiple Reaction Monitoring mode with precursor-to-product ion transitions at m/z of 229 to m/z of 114 as MH+ion for Decitabine, m/z of 269 to m/z of 118 as MH+ion for Cedazuridine. A very sensitive limit of detection of 0.3 ng/ml was observed and showed a calibration curve linear over the concentration range of LLOQ (lower limit of quantification) to 500 ng/ml. The other validation parameters were found to have acceptable accuracy, precision, linearity, and selectivity. The mean extraction concentration was acceptable and very high for both the analytes in HQC (high-quality control concentration), MQC (medium quality control concentration) and LLOQ levels. The peak area response ratio of Decitabine and Cedazuridine with the internal standard in freeze-thaw, short term and long term stability studies was found to be acceptable confirms that the method is stable. Conclusion: It can be concluded that the proposed method is specific, accurate, and precise and could be used for the simultaneous estimation of Decitabine and Cedazuridine in human plasma.

Download Full-text