scholarly journals Development and Validation of an HPLC Method for Quantification of Filgotinib, a Novel JAK-1 Inhibitor in Mice Plasma: Application to a Pharmacokinetic Study

Drug Research ◽  
2020 ◽  
Vol 70 (05) ◽  
pp. 233-238
Author(s):  
Ashok Zakkula ◽  
Shobha Pulipati ◽  
Sreekanth Dittakavi ◽  
Ram Murthi Bestha ◽  
Mohd Zainuddin ◽  
...  
Keyword(s):  
Pharmacokinetic Study ◽  
High Performance ◽  
Kinase Inhibitor ◽  
Internal Standard ◽  
Hplc Method ◽  
Janus Kinase ◽  
Extraction Solvent ◽  
Long Term Storage ◽  
Development And Validation

AbstractFilgotinib is a selective JAK1 (Janus kinase) inhibitor, filed in Japan for the treatment of rheumatoid arthritis. In this paper, we present the data of development and validation of a high-performance liquid chromatography (HPLC) method for the quantitation of filgotinib in mice plasma as per the FDA regulatory guideline. The method involves the extraction of filgotinib along with internal standard (IS, tofacitinib) from mice plasma (100 µL) using ethyl acetate as an extraction solvent. The chromatographic analysis was performed using an isocratic mobile phase comprising 10 mM ammonium acetate (pH 4.5) and acetonitrile (70:30, v/v) at a flow-rate of 0.8 mL/min on a Hypersil Gold C18 column. The UV detection wavelength was set at λmax 300 nm. Filgotinib and the IS eluted at 5.56 and 4.28 min, respectively with a total run time of 10 min. The calibration curve was linear over a concentration range of 0.05 to 5.00 μg/mL (r 2+=≥0.992). The intra- and inter-day precision and accuracy results were within the acceptable limits. Results of stability studies indicated that filgotinib was stable on bench-top, in auto-sampler, up to three freeze/thaw cycles and long-term storage at −80°C. The validated HPLC method was successfully applied to a pharmacokinetic study in mice.

Validated HPLC Method for Quantification of a Novel Trk Inhibitor, Larotrectinib in Mice Plasma: Application to a Pharmacokinetic Study

Drug Research ◽  
2020 ◽  
Vol 70 (02/03) ◽  
pp. 101-106
Author(s):  
Harsha K. Tripathy ◽  
S.V. Nair Manju ◽  
Ashok Zakkula ◽  
Ram Murthi Bestha ◽  
Sreekanth Dittakavi ◽  
...  
Keyword(s):  
Pharmacokinetic Study ◽  
High Performance ◽  
Wave Length ◽  
Internal Standard ◽  
Hplc Method ◽  
Long Term Storage ◽  
Development And Validation ◽  
Orally Active ◽  
Regulatory Guideline

AbstractLarotrectinib, is an orally active novel small molecule approved for the treatment of solid tumors in pediatrics and adult patients. It acts by inhibiting tropomyosin receptor kinase. In this paper, we report the development and validation of a high-performance liquid chromatography (HPLC) method for the quantitation of larotrectinib in mice plasma as per the FDA regulatory guideline. Plasma samples processing was accomplished through simple protein precipitation using acetonitrile enriched with internal standard (IS, enasidenib). The chromatographic analysis was performed using a gradient mobile phase comprising 10 mM ammonium acetate and acetonitrile at a flow-rate of 0.8 mL/min on an X-Terra Phenyl column. The UV detection wave length was set at λmax 262 nm. Larotrectinib and the IS eluted at 3.85 and 6.60 min, respectively with a total run time of 8.0 min. The calibration curve was linear over a concentration range of 0.20–5.00 μg/mL (r2=≥0.992). The intra- and inter-day precision and accuracy results were within the acceptable limits. Results of stability studies indicated that larotrectinib was stable on bench-top, in auto-sampler, up to three freeze/thaw cycles and long-term storage at −80°C. The validated HPLC method was successfully applied to a pharmacokinetic study in mice.

scholarly journals Validated HPLC method for quantification of copanlisib in mice plasma: application to a pharmacokinetic study

ADMET & DMPK ◽  
2020 ◽  
Vol 8 (1) ◽  
pp. 113-121
Author(s):  
Ashok Zakkula ◽  
Pavan Kumar Kurakula ◽  
Sreekanth Dittakavi ◽  
Prasanthi Daram ◽  
Ram Murthi Bestha ◽  
...  
Keyword(s):  
Pharmacokinetic Study ◽  
High Performance ◽  
Internal Standard ◽  
Hplc Method ◽  
Array Detector ◽  
Extraction Solvent ◽  
Long Term Storage ◽  
Thaw Cycle ◽  
Freeze Thaw Cycle ◽  
Development And Validation

Copanlisib is a pan phosphatidylinositol 3-kinase (PI3K) inhibitor approved for follicular lymphoma. In this paper, we present the data of development and validation of a high-performance liquid chromatography (HPLC) method for the quantitation of copanlisib in mice plasma as per the FDA regulatory guideline. The method involves the extraction of copanlisib along with internal standard (IS, enasidenib) from mice plasma (100 µL) using ethyl acetate as an extraction solvent. The chromatographic resolution of copanlisib and the IS was achieved on a Hypersil Gold C18 column maintained at 40 °C using a binary gradient mobile phase [10 mM ammonium formate (pH 4.0) and acetonitrile]. The flow-rate was 0.8 mL/min. For the detection of copanlisib and the IS, the photo-diode array detector was set at λmax 310 nm. Copanlisib and the IS eluted at 6.60 and 7.80 min, respectively with a total run time of 10 min. The calibration curve was linear over a concentration range of 50 to 5000 ng/mL for copanlisib (r2³ 0.998). The results of intra- and inter-day accuracy and precision studies were within the acceptable limits. Copanlisib was stable on bench-top, in auto-sampler, up to three freeze/thaw cycle and long-term storage at -80 °C. The application of the validated method was shown in a mice pharmacokinetic study.

scholarly journals Validated DBS method for filgotinib quantitation in rat dried blood spots and its application to a pharmacokinetic study in rats

ADMET & DMPK ◽  
2020 ◽  
Author(s):  
Abhishek Dixit ◽  
Vinay Kiran ◽  
Bhavesh Babulal Gabani ◽  
Ramesh Mullangi
Keyword(s):  
Pharmacokinetic Study ◽  
Kinase Inhibitor ◽  
Plasma Concentrations ◽  
Internal Standard ◽  
Dried Blood Spots ◽  
Janus Kinase ◽  
Extraction Solvent ◽  
Validation Parameters ◽  
Carry Over ◽  
Development And Validation

<p class="ADMETabstracttext">Filgotinib is a selective JAK1 (Janus kinase) inhibitor, showed efficacy in patients suffering from moderate-to-severe rheumatoid arthritis. In this paper, we present the data on the development and validation of a sensitive, selective and high-throughput LC-MS/MS (liquid chromatography with tandem mass spectrometry) method for the quantitation of filgotinib from rat dried blood spot (DBS) cards. To the DBS disc cards, 0.2 % formic acid enriched with internal standard (IS) was added and sonicated. Thereafter the extraction of filgotinib and the IS (tofacitinib) was accomplished using ethyl acetate as an extraction solvent. The resolution of filgotinib and the IS was achieved on a Gemini C<sub>18</sub> column with an isocratic<strong> </strong>mobile phase, which is a mixture of 0.2 % formic acid:acetonitrile (20:80, v/v) at a flow-rate of 0.9 mL/min. The total run time was 2.90 min and the retention time of filgotinib and the IS was ~1.31 and 0.89 min, respectively. Filgotinib and the IS were analyzed using positive ion scan mode and parent-daughter mass to charge ion (m/z) transition of 426.3®291.3 and m/z 313.2®149.2, respectively, for quantitation. The calibration range was 1.37-1937 ng/mL. No matrix effect and carry over were observed. All the validation parameters met the acceptance criteria. The validated method has been applied to a pharmacokinetic study in rats. A good correlation between DBS and plasma concentrations for filgotinib was observed.</p>

Development and validation of a LC-MS/MS method for quantification of mobocertinib (TAK-788) in plasma and its application to pharmacokinetic study in rats

2020 ◽  
Vol 23 ◽  
Author(s):  
Bo Li ◽  
Jin Wang ◽  
Xinyao Dou ◽  
Xinjie Zhang ◽  
Xianbei Xue ◽  
...  
Keyword(s):  
Oral Administration ◽  
Pharmacokinetic Study ◽  
High Performance ◽  
Kinase Inhibitor ◽  
Internal Standard ◽  
Selected Reaction Monitoring ◽  
Precipitation Method ◽  
Plasma Samples ◽  
Bioanalytical Method Validation ◽  
Extraction Recovery

Aim and Objective:: An analytical method for the determination of mobocertinib, an investigational tyrosine kinase inhibitor, was developed and optimized by high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) in rat plasma. Materials and Methods:: Plasma samples were pretreated by the protein precipitation method with a methanol solution of osimertinib as the internal standard (IS). Chromatographic separation was performed using an Inertsil ODS-3 column (50 mm × 4.6 mm, I.D. 5 μm) column with the temperature maintained at 40 °C. The mobile phase consisted of water (containing 0.1% formic acid) and methanol in a gradient mode at a flow rate of 0.5 mL/min. Mass spectrometric detection was carried out in the selected reaction monitoring (SRM) mode with positive electrospray ionization, and the mass transitions of mobocertinib and osimertinib were m/z 587.01 → 71.88 and m/z 499.80 → 71.94, respectively. The method was validated in terms of selectivity, linearity, accuracy and precision, extraction recovery and matrix effect, stability and carryover as per the guidelines for bioanalytical method validation (FDA, 2018). The method was applied to the pharmacokinetic study of mobocertinib in rats by oral gavage at the doses of 2, 6, and 18 mg/kg. A total of 216 plasma samples from 18 rats were analyzed. Results:: It showed good linearity over the range of 1-1000 ng/mL (R2 = 0.9957). The intra-batch accuracy was within 94.65-102.59% and the precision was within 5.49-10.46%. The inter-batch accuracy was within 97.08-102.25% with a precision of 7.54-10.13%. The extraction recovery and matrix factor were acceptable for the bioanalysis of mobocertinib. Additionally, mobocertinib was found to be stable under the detected conditions. Mobocertinib showed linear pharmacokinetic characteristics following oral administration to rats at 2.0-18.0 mg/kg. Conclusion:: The developed and validated method was successfully employed in the pharmacokinetic study in rats following oral administration of mobocertinib at the doses of 2, 6, and 18 mg/kg.

scholarly journals Development and validation of a rapid UHPLC-MS/MS method for the determination of fenofibric acid in human plasma: Application to a pharmacokinetic study of fenofibrate tablet in Chinese subjects

2020 ◽  
Author(s):  
Xiaorong Wu ◽  
Yankai Wang ◽  
Binbin Liang ◽  
Honghai Wu ◽  
Liying Wu ◽  
...  
Keyword(s):  
Human Plasma ◽  
Pharmacokinetic Study ◽  
High Performance ◽  
Multiple Reaction Monitoring ◽  
Internal Standard ◽  
Fenofibric Acid ◽  
The Matrix ◽  
Liquid Chromatography Tandem Mass ◽  
Development And Validation ◽  
Chinese Subjects

AbstractAn ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC–MS/MS) method was developed to determine the fenofibric acid (FA) in human plasma and applied to a pharmacokinetic study of fenofibrate tablet (Lipanthyl® supra, 160 mg) on Chinese subjects which had not been reported. Bezafibrate was used as an internal standard (IS), and the plasma samples were precipitated by methanol. Multiple reaction monitoring (MRM) mode was used to quantitatively analyzed FA m/z 317.2 → 230.7 and the IS m/z 360.0 → 274.0 in the electrospray ionization (ESI) negative interface. The calibration curves were linear over the range of 50–30,000  ng/mL (r2  ≥  0.996). The intra-day and inter-day precision (coefficient of variation, CV%) was less than 2.7 and 2.5%, respectively. The accuracy (relative error, RE%) ranged from −4.5 to 6.9%. The average recovery was higher than 86.2%, and the matrix effect was between 95.32 and 110.55%. The simple, rapid, and selectivity method was successfully applied to the pharmacokinetic study of fenofibrate tablets on Chinese subjects.

Development and Validation of Midostaurin Assay by RP-HPLC Method

2018 ◽  
Vol 11 (6) ◽  
pp. 4318-4322
Author(s):  
Abrar Ahmed ◽  
Tayyaba Mahtab ◽  
Sumaiyya Saleem
Keyword(s):  
Myeloid Leukemia ◽  
High Performance ◽  
Kinase Inhibitor ◽  
Pharmaceutical Formulations ◽  
High Performance Liquid Chromatographic ◽  
Hplc Method ◽  
Liquid Chromatographic Method ◽  
Development And Validation ◽  
Liquid Chromatographic

Midostaurin is a multi-targeted protein kinase inhibitor that has been used for the treatment of acute myeloid leukemia.  Here, a rapid and precise reverse phase high-performance liquid chromatographic method has been developed for the validation of midostaurin, in its API form as well as in capsule dosage form. Chromatography was carried out on a X-Bridge C18 (4.6 x 250 mm, 5 µm) column using a mixture of methanol: water (75:25% v/v) as the mobile phase at a flow rate of 1.0 mL/min, the detection was carried out at 243nm and the retention time of the midostaurin was found to be 3.155. The method produce linear responses in the concentration range of 10-50 µg/mL of midostaurin. The method precision for the determination of assay was below 2.0 % RSD. The LOD and LOQ values obtained were 1.2 µg/mL and 3.8 µg/mL respectively. There were no significant changes observed upon changing chromatographic conditions indicating the method to be robust. Therefore this validated method can be useful in the quality control of bulk and pharmaceutical formulations of midostaurin. 

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

INDIAN DRUGS ◽  
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.

Development and Validation of HPLC-UV Method for the Determination of a Potent Synthetic Cannabinoid THJ-2201 in Mouse Plasma and Application in a Pharmacokinetic Study

2020 ◽  
Vol 16 (4) ◽  
pp. 404-411
Author(s):  
Hassan Y. Aboul-Enein ◽  
Gamal A.E Mostafa ◽  
Haitham AlRabiah ◽  
Mohammed Al-Ramadi ◽  
Sabry M. Attia ◽  
...  
Keyword(s):  
Pharmacokinetic Study ◽  
High Performance ◽  
Limit Of Detection ◽  
Internal Standard ◽  
Reversed Phase ◽  
Hplc Method ◽  
Synthetic Cannabinoid ◽  
Mean Values ◽  
Mouse Plasma

Aim: A new simple and sensitive high-Performance Liquid Chromatography (HPLC) method for the determination of a potent synthetic cannabinoid THJ-2201, has been developed and validated. Lixiviptan was used as the Internal Standard (IS). Methods: THJ-2201 and IS were extracted from mouse plasma using deproteinization procedure that uses acetonitrile followed by HPLC analysis. The separation was carried out on a reversed-phase C18 column using water and acetonitrile mixture (30:70 v/v). The flow-rate was 1.0 mL/min. Eluting of both THJ-2201 and lixivaptan was performed at 220 nm. Results: The method demonstrated linearity over a calibration range of 95 - 1500 ng/mL and the Limit of Detection (LOD) and Quantitation (LOQ) were 28 ng/mL and 91 ng/mL, respectively. The validation of the proposed method was carried out by following the US Food and Drug Administration (FDA) guidelines. Intra- and inter-day precision did not exceed 6.4%, whereas the accuracy of THJ-2201 measurements was within ±13%. Conclusion: This new method is simple and sensitive and has been applied successfully in a pharmacokinetic study of THJ-2201 in mouse plasma. The mean values of Tmax and Cmax were 0.25 h and 141.87 ± 12.11 ng/mL, respectively.

Development and Application of HPLC Method for Clinical Pharmacokinetic Study of Domperidone

2016 ◽  
Vol 9 (6) ◽  
pp. 3531-3535
Author(s):  
Prasad Neerati ◽  
Bhargavi Latha A ◽  
Y. Shravan Kumar ◽  
Y Madhusudan Rao
Keyword(s):  
Pharmacokinetic Study ◽  
Mobile Phase ◽  
High Performance ◽  
Internal Standard ◽  
High Performance Liquid Chromatographic ◽  
Hplc Method ◽  
Assay Method ◽  
Uv Detector ◽  
Liquid Chromatographic Method ◽  
Liquid Chromatographic

A simple and sensitive high performance liquid chromatographic method for quantification of domperidone in human serum was developed and validated. Domperi-done and internal standard (IS) propranolol hydrochloride were extracted into acetonitrile and separated using an isocratic mobile phase on a Phenomenx C18 column. The eluent was monitored by UV detector at 270 nm at a flow rate of 1.0 mL min–1. The linearity range of proposed  method was 2.5–1000 ng/ml. The intra-day and inter-day coefficient of variation and percent error values of the assay method were less than 15% and mean recovery was more than 97 and 96% for domperidone and IS, respectively. The method was found to be precise, accurate and specific during the study. The method was successfully applied for pharmacokinetic study of domperidone in humans.  

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