Development and Validation of an UPLC-MS/MS method for the Simultaneous determination of Verapamil and Trandolapril in Rat Plasma: Application to a Pharmacokinetic Study

Background: Verapamil is an excellent drug for the medication of hypertension and Trandolapril is an angiotensin-converting-enzyme inhibitor. So, it is an interesting method to develop a novel and reliable MS/UPLC strategy for simultaneous establishment of Verapamil and Trandolapril. Objective: To develop a new rapid and sensitive UPLC-MS/MS method for the simultaneous estimation of Verapamil and Trandolapril in rat plasma using D6-Verapamil and D6-Trandolapril. Method: Separation was carried on column Symmetry C18 column (150x4.6mm, 3.5µm) using a isocratic elution with a buffer containing 1mL of Formic acid in 1Lit of water and the mixture of two components like Buffer and Acetonitrile in the ratio of 80:20 as mobile phase with 1mL/min flow rate at ambient temperature. Results: Analysis was performed within 5 minutes over a good linear concentration range from 2.4ng/mL to 48ng/mL(r2 = 0.9993 ± 0.018) for Verapamil and 10pg/mL to 200pg/mL(r2 =0.9993± 0.006) for Trandolapril .The extraction recoveries and matrix effect of verapamil and Trandolapril were 98.45,99.95, 98.12, 99.66% and 98.27,99.89, 97.78, 99.23% at different QC concentration levels. Precision and recovery study results are within the acceptable limit. An electro spray ionization source was used to study verapamil and Trandolapril at m/z 454.72→182.16, 430.25→201.48 and IS for m/z 460.18→ 324.39, 436.28 → 340.52 were ion pairs of mass analysis. This method has been successfully applied, exploring Verapamil (1.2mg/kg) with its internal standard (D6-Verapamil), Trandolapril (0.005mg/kg) with its internal standard (D6-Trandolapril) were extracted from rat plasma using liquid -liquid extraction. Conclusion: This manuscript focuses on the consistent evaluation of the key Bioanalytical validation parameters and the following are discussed: accuracy, precision, sensitivity, selectivity, standard curve, limits of quantification, range, recovery, and stability. These validation parameters are described, together with illustrations of validation methodology applied in the case of chromatographic methods used in bio analysis.

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Quantitation of capmatinib, a mesenchymal-epithelial transition factor inhibitor by UPLC–MS/MS in rat plasma and its application to a pharmacokinetic study

Bioanalysis ◽

10.4155/bio-2020-0011 ◽

2020 ◽

Vol 12 (5) ◽

pp. 285-293

Author(s):

Chunling Zhou ◽

Jinmiao Tian ◽

Peng Lin ◽

Tianzhu Liu ◽

Aiping He ◽

...

Keyword(s):

Pharmacokinetic Study ◽

Ion Pairs ◽

Internal Standard ◽

Rat Plasma ◽

Multiple Cancer ◽

Factor Inhibitor ◽

Orally Bioavailable ◽

Mesenchymal Epithelial Transition Factor ◽

Acquity Uplc ◽

Mesenchymal Epithelial Transition

Aim: Capmatinib is an orally bioavailable mesenchymal-epithelial transition factor inhibitor with anticancer activity, which has proved preclinical activity in multiple cancer trials. The present study aimed to develop a fast and reliable assay approach to quantify capmatinib in rat plasma. Methodology & results: After protein precipitation with acetonitrile, the chromatographic separation was achieved with an Acquity UPLC BEH C18 column, and subsequently detected with positive electrospray ionization via a triple quadrupole tandem mass spectrometer. The target quantitative ion pairs m/z 412.99 → 381.84 for capmatinib and 387.00 → 355.81 for the internal standard, respectively. The calibration curve for the assay was linear over the range of 1.0–4000 ng/ml. Conclusion: The method shows an excellent performance in linearity, accuracy, precision, stability, and has been successfully applied to a pharmacokinetic study after oral administration of capmatinib at three doses (5, 10 and 20 mg/kg) in rats.

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Pharmacokinetic Comparison of Isoalantolactone and Alantolactone in Rats after Administration Separately by Optimization of an UPLC-MS2Method

Journal of Chemistry ◽

10.1155/2014/354618 ◽

2014 ◽

Vol 2014 ◽

pp. 1-8 ◽

Cited By ~ 3

Author(s):

Renjie Xu ◽

Mengyue Wang ◽

Ying Peng ◽

Xiaobo Li

Keyword(s):

Multiple Reaction Monitoring ◽

Internal Standard ◽

Rat Plasma ◽

Sprague Dawley ◽

Ionization Source ◽

Fragment Ions ◽

Sprague Dawley Rats ◽

Liquid Chromatography Tandem Mass ◽

Positive Ion

Isoalantolactone and alantolactone are two major active ingredients that are present in many medicinal plants. In this study, a sensitive and rapid ultraperformance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed for determination of the two compounds in rat plasma, separately. In this method, an electrospray ionization source was applied and operated in positive ion mode; multiple reaction monitoring (MRM) was selected for quantification using target fragment ions 233.2→187.1 for isoalantolactone (alantolactone) and 245.1→189.1 for internal standard (IS). Retention time of the lactones and IS was within 3.0 min. Further calibration suggested a linear regression can be calculated within 2.5–500 ng/mL for isoalantolactone and 4–500 ng/mL for alantolactone. This method was used to compare the pharmacokinetic characteristics of isoalantolactone and alantolactone at a single dose of 5 mg/kg into male Sprague-Dawley rats by intravenous administration separately. The levels oft1/2, Kel, CL,Cmax, and AUC were significantly increased in the alantolactone group compared to isoalantolactone. These results suggested that isoalantolactone was distributed and eliminated more rapidly than alantolactone in rats when administered, respectively.

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DEVELOPMENT AND VALIDATION OF NEW RP–HPLC METHOD FOR DETERMINATION OF BESIFLOXACIN IN ANIMAL MODEL

INDIAN DRUGS ◽

10.53879/id.52.01.10228 ◽

2015 ◽

Vol 52 (01) ◽

pp. 26-32

Author(s):

A Singh ◽

◽

C. L Singh ◽

S. Kumar ◽

M. Kumar

Keyword(s):

High Performance ◽

Internal Standard ◽

Rat Plasma ◽

Reversed Phase ◽

Hplc Method ◽

Standard Curve ◽

Absorbance Detection ◽

Rp Hplc ◽

Ich Guidelines ◽

Development And Validation

A sensitive and accurate reversed– phase high performance liquid chromatography (RP-HPLC) method with UV absorbance detection at 289 nm was developed and validated for the determination and quantification of besifloxacin (BSF) in rat plasma. Ofloxacin was used as an internal standard (IS). The sample was prepared by liquid extraction of BSF from plasma, using methanol and acetonitrile (70:30). The chromatographic separation was achieved with octadecylsilane (ODS-3), Hypersil® C18 column (250 mm×6mm×5μm). The chromatographic runtime was less than 5 minutes where the retention time of internal standard and the drug was 2.15 min and 3.30 min respectively. A standard curve with a regression coefficient (r2) 0.999 was obtained in the range of 0.025-20 μg/mL. The method was validated with respect to linearity, range, precision, accuracy and robustness according to ICH guidelines. The method was found to be accurate and robust with a runtime of less than 5 minutes. Hence, the present method was rapid and economical to use for clinical studies as well as to analyze the drug in different plasma samples.

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Simultaneous determination of two galangin metabolites from Alpinia Officinarum Hance in rat plasma by UF LC-MS/MS and its application in pharmacokinetics study

PeerJ ◽

10.7717/peerj.11041 ◽

2021 ◽

Vol 9 ◽

pp. e11041

Author(s):

Rangru Liu ◽

Hailong Li ◽

Na Wei ◽

Yinfeng Tan

Keyword(s):

Analytical Method ◽

Glucuronic Acid ◽

Multiple Reaction Monitoring ◽

Ion Pairs ◽

Internal Standard ◽

Rat Plasma ◽

Pharmacokinetic Parameters ◽

Alpinia Officinarum ◽

Negative Electrospray Ionization

Galangin has multiple pharmacological efficacies, such as anti-cancer, anti-inflammation and anti-oxidation. Galangin can be rapidly converted into glucuronidated metabolites in vivo. This study aimed to establish an UFLC-MS/MS analytical method to simultaneously determine the concentrations of two glucuronidated metabolites of galangin, galangin-3-O-β-D-glucuronic acid (GG-1) and galangin-7-O-β-D-glucuronic acid (GG-2) in rat plasma. After oral administration of galangal extract (0.3 g/kg), blood samples were collected from the orbital sinus, then treated by methanol precipitation and further gradient-eluted with Phenomenex Kinetex 2.6 µm XB-C18 column. The mass spectrometer was manipulated in the negative electrospray ionization (ESI) and selected multiple reaction monitoring (MRM) mode for the analytes. The precursor-to-product ion pairs applied for GG-1, GG-2 and chrysin (as the internal standard, IS) were m/z 445.2→269.0, 445.2→268.9 and 253.0→142.9, respectively. The results showed that the linear ranges for both GG-1 and GG-2 were 2.0–2000.0 ng/mL (r2 > 0.995). The inter- and intra-day precision were 89.3%–109.2%, RSD was less than 15%, and the repeatability was good. The recoveries of both metabolites and IS were over 89%, and matrix effect was within 15%. The validated analytical method was further applied to study the pharmacokinetic profiles of GG-1 and GG-2 in vivo. The pharmacokinetic parameters suggested that Tmax of GG-1 was equivalent to that of GG-2, and MRT0-t, t1/2 of GG-2 were a little higher than those of GG-1. Importantly, AUC0-t and Cmax of GG-2 were almost twice as those of GG-1. In short, the validated UFLCMS/MS analytical method was feasible to simultaneously determine two galangin metabolites GG-1 and GG-2 in rat plasma and further analyze in vivo metabolism of galangin.

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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.

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Pharmacokinetic Drug-Drug Interaction and Responsible Mechanism between Memantine and Cimetidine

Pharmaceutics ◽

10.3390/pharmaceutics10030119 ◽

2018 ◽

Vol 10 (3) ◽

pp. 119

Author(s):

Young Choi ◽

Im-Sook Song ◽

Min-Koo Choi

Keyword(s):

Drug Interaction ◽

Drug Interactions ◽

Intestinal Permeability ◽

Internal Standard ◽

Rat Plasma ◽

Precipitation Method ◽

Plasma Exposure ◽

Standard Curve ◽

Drug Drug Interaction ◽

Pharmacokinetic Drug

A sensitive and simple chromatography-tandem mass spectrometry (LC-MS/MS) method was developed to evaluate memantine in rat plasma. Memantine and propranolol (internal standard) in rat plasma was extracted using a methanol precipitation method. The standard curve value was 0.2–1000 ng/mL and selectivity, linearity, inter-day and intra-day accuracy and precision were within acceptance criteria. Using this validated method, drug-drug interactions between memantine and cimetidine was measured following co-administration of memantine and cimetidine intravenously and orally. Plasma exposure of memantine was increased by 1.6- and 3.0-fold by co-medication with cimetidine intravenously and orally, respectively. It suggested that the drug interaction occurred during the gut absorption process, which was consistent with the results showing that the intestinal permeability of memantine in the presence of cimetidine was 3.2-fold greater than that of memantine alone. Inhibition of cimetidine on hepatic elimination of memantine rather than renal excretion was also attributed to the drug-drug interaction between memantine and cimetidine, which explained the decreased clearance of memantine by co-medication with cimetidine. In conclusion, the newly developed simple and sensitive LC-MS/MS analytical method was applied to investigate the pharmacokinetic drug-drug interactions of memantine. Plasma exposure of memantine by co-administration with cimetidine was increased because of its enhanced intestinal permeability and the decreased metabolic activity of memantine.

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A Rapid UPLC-MS Method for Quantification of Gomisin D in Rat Plasma and Its Application to a Pharmacokinetic and Bioavailability Study

Molecules ◽

10.3390/molecules24071403 ◽

2019 ◽

Vol 24 (7) ◽

pp. 1403 ◽

Cited By ~ 2

Author(s):

Xiaoyong Zheng ◽

Feng Feng ◽

Xiunan Jiang ◽

Jieying Qiu ◽

Xiaojun Cai ◽

...

Keyword(s):

Multiple Reaction Monitoring ◽

Internal Standard ◽

Ultra Performance Liquid Chromatography ◽

Rat Plasma ◽

Ionization Source ◽

Quality Marker ◽

Liquid Chromatography Tandem Mass ◽

Bioavailability Study ◽

Multiple Reaction Monitoring Mode ◽

Positive Ion

Gomisin D, a lignan compound isolated from Fructus Schisandra, is a potential antidiabetic and anti-Alzheimer’s agent. Recently, gomisin D was used as a quality marker of some traditional Chinese medicine (TCM) formulas. In this study, a rapid ultra-performance liquid chromatography/tandem mass spectrometry method (UPLC-MS/MS) was developed and validated to quantify gomisin D in rat plasma for a pharmacokinetic and bioavailability study. Acetonitrile was used to precipitate plasma proteins. Separations were performed on a BEH C18 column with a gradient mobile phase comprising of acetonitrile and water (0.1% formic acid). An electrospray ionization source was applied and operated in the positive ion mode. The multiple reaction monitoring mode (MRM) was utilized to quantify gomisin D and nomilin (internal standard, IS) using the transitions of m/z 531.2 → 383.1 and m/z 515.3 → 161.0, respectively. The calibration curve was linear over the working range from 1 to 4000 ng/mL (R2 = 0.993). The intra- and interday precision ranged from 1.9% to 12.9%. The extraction recovery of gomisin D was in the range of 79.2–86.3%. The validated UPLC-MS/MS method was then used to obtain the pharmacokinetic characteristics of gomisin D after intravenous (5 mg/kg) and intragastric (50 mg/kg) administration to rats. The bioavailability of gomisin D was 107.6%, indicating that this compound may become a promising intragastrical medication. Our results provided useful information for further preclinical studies on gomisin D.

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Development and Validation of UPLC-MS/MS Method for the Determination of Aripiprazole in Rat Plasma Using Liquid-Liquid Extraction: Pharmacokinetic and Bioequivalence Application

Asian Journal of Chemistry ◽

10.14233/ajchem.2020.22899 ◽

2020 ◽

Vol 32 (10) ◽

pp. 2671-2676

Author(s):

Ashish Raghuvanshi ◽

Urooj A. Khan ◽

Uzma Parveen ◽

Anshul Gupta ◽

Gaurav K. Jain

Keyword(s):

Internal Standard ◽

Rat Plasma ◽

Liquid Extraction ◽

Single Step ◽

Liquid Liquid Extraction ◽

Validation Parameters ◽

Tandem Mass Spectrometric ◽

Liquid Chromatographic

A selective, simple, sensitive and rapid ultra-performance liquid chromatographic tandem mass spectrometric (UPLC-MS/MS) method for the detection of aripiprazole in rat plasma has been developed and validated using aripiprazole-D8 as internal standard (IS). A simple single step sample preparation process was accomplished by liquid-liquid extraction (LLE). The post-treatment samples were chromato-graphed and analyzed on a UPLC bridged ethyl hybrid (BEH) C-18 column using mobile phase composition of acetonitrile: 0.1% formic acid in water::70:30 (v/v). Aripiprazole was analyzed by MS detector in positive electrospray ionization mode (ESI). Multiple reactions monitoring (MRM) was employed to observed the transition for aripiprazole (m/z 448.35→285.09) and aripiprazole-D8 (m/z 456.2→293.2). The developed method was validated and found linear in the working range of 2-1025 ng/mL with correlation coefficient, r2 = 0.99951 and quantification limit of 2.02 ng/mL. All validation parameters were in accordance with the ICH guidelines and met the acceptance criteria. The method was found to be accurate (recovery, 97.07 to 103.64%, precise (% CV, 2.68 to 7.70%), rapid (run time 4 min) and specific. The validated method was successfully used for the determination of plasma concentration of aripiprazole after single oral administration in rats and hence could be useful for in vivo pharmacokinetic study and bioequivalence testing of aripiprazole formulations.

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Comparative Pharmacokinetic Study of Taxifolin after Oral Administration of Fructus Polygoni Orientalis Extract in Normal and Fibrotic Rats by UPLC-MS/MS

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2019/9348075 ◽

2019 ◽

Vol 2019 ◽

pp. 1-10 ◽

Cited By ~ 1

Author(s):

Feili Wei ◽

Li Guo ◽

Yongsong Xu ◽

Dexi Chen ◽

Muxin Gong

Keyword(s):

Liver Fibrosis ◽

Oral Administration ◽

Residence Time ◽

Mean Residence Time ◽

Internal Standard ◽

Rat Plasma ◽

Pharmacokinetic Parameters ◽

Porcine Serum ◽

Validation Parameters ◽

The Mean

Fructus polygoni orientalis (FPO) is widely used in clinical practice in China, especially in treatment of liver diseases including viral hepatitis, liver fibrosis, and liver cirrhosis. However, its pharmacokinetic (PK) alterations in liver fibrotic rats have rarely been reported. To study whether taxifolin, one of the main flavonoids in FPO can be absorbed into blood after oral administration of FPO extract and to compare the differences in pharmacokinetic parameters of taxifolin to normal and liver fibrotic rats induced by porcine serum (PS), a UPLC-MS/MS method was developed and validated for determination of taxifolin in rat plasma using puerarin as the internal standard (IS). All validation parameters met the acceptance criteria according to regulatory guidelines. The results indicated that after treatment of rats with PS alone for 12 weeks, the liver fibrotic model group was built successfully. The taxifolin can be absorbed into the blood after oral administration of the FPO extract. The Cmax of taxifolin was 1940 ± 502.2 ng/mL and 2648 ± 208.5 ng/mL (p<0.05), the AUC0∼t of taxifolin was 4949.7 ± 764.89 h·ng/mL and 6679.9 ± 734.26 h·ng/mL (p<0.05), the AUC0∼∞ of taxifolin was 5049.4 ± 760.7 and 7095.2 ± 962.3 h·ng/mL (p<0.05), and the mean residence time (MRT) of taxifolin was 2.46 ± 0.412 h and 3.17 ± 0.039 h (p<0.05) in the normal and fibrotic model groups, respectively. These results confirmed that the pharmacokinetic parameters of taxifolin are altered in liver fibrosis, manifested as Cmax, AUC0∼t, AUC0∼∞, and the mean residence time (MRT). It suggested that it is essential to consider the characteristics of pharmacokinetics after oral administration of FPO in liver disease patients.

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Quick and Sensitive UPLC-ESI-MS/MS Method for Simultaneous Estimation of Sofosbuvir and Its Metabolite in Human Plasma

Molecules ◽

10.3390/molecules24071302 ◽

2019 ◽

Vol 24 (7) ◽

pp. 1302 ◽

Cited By ~ 1

Author(s):

Mohammad Semreen ◽

Hasan Alniss ◽

Muath Mousa ◽

Hassan Aboul-Enein

Keyword(s):

Human Plasma ◽

Matrix Effects ◽

Multiple Reaction Monitoring ◽

Internal Standard ◽

Simultaneous Estimation ◽

High Throughput Analysis ◽

Elution Time ◽

Validation Parameters ◽

Multiple Reaction Monitoring Mode ◽

Acquity Uplc

A simple, fast and highly sensitive RP-UPLC-MS/MS method was developed and validated for the simultaneous determination of sofosbuvir (SR) and its metabolite GS331007 in human plasma using ketotifen as an internal standard (IS). The separation was achieved on Acquity UPLC BEH C18 (50 × 2.1 mm, i.d. 1.7 µm, Waters, USA) column using acetonitrile:5 mM ammonium formate:0.1% formic acid (85:15:0.1% v/v/v) as a mobile phase at a flow rate of 0.35 mL/min in an isocratic elution. The Xevo TQD UPLC-MS/MS was operated under the multiple-reaction monitoring mode using positive electrospray ionization. Extraction with dichloromethane was used in the sample preparation. Method validation was performed as per the Food and Drug Administration (FDA) guidelines and the calibration curves of the proposed method were found to be linear in the range of 1–1000 ng/mL for SR and in the range of 10–1500 ng/mL for its metabolite (GS331007) with an elution time of 1.83 min. All validation parameters were within the acceptable range according to the bioanalytical methods validation guidelines. Furthermore, the obtained results of matrix effects indicate that ion suppression or enhancement from human plasma components was negligible under the optimized conditions. The proposed method can be applied in high-throughput analysis required for pharmacokinetic and bioequivalence studies in human samples.

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