Development of a Simple and Validated LC–MS/MS Method for Quantitative Determination of Ketotifen in Beagle Dog Plasma and Its Application to Bioequivalence Study of Ketotifen Syrup Dosage Form

Song, Eunseo; Shim, Wang-Seob; Choi, Doowon; Song, Yuna; Jo, Hyeong Geun; Lee, Soobok; Jung, Suk Han; Choi, Yeo Jin; Lee, Kyung-Tae

doi:10.3390/molecules29184505

Open AccessArticle

Development of a Simple and Validated LC–MS/MS Method for Quantitative Determination of Ketotifen in Beagle Dog Plasma and Its Application to Bioequivalence Study of Ketotifen Syrup Dosage Form

by

Eunseo Song

^1,2,3

,

Wang-Seob Shim

²

,

Doowon Choi

^1,2,3,

Yuna Song

^1,2,3,

Hyeong Geun Jo

⁴,

Soobok Lee

^4,5,

Suk Han Jung

^4,6,

Yeo Jin Choi

^3,*

and

Kyung-Tae Lee

^1,2,3,*

¹

Department of Biomedical and Pharmaceutical Science, Graduated School, Kyung Hee University, Seoul 02447, Republic of Korea

²

Kyung Hee Drug Analysis Center, College of Pharmacy, Kyung Hee University, Seoul 02447, Republic of Korea

³

Department of Pharmacy, College of Pharmacy, Kyung Hee University, Seoul 02447, Republic of Korea

⁴

Daewon Pharm. Co., Ltd., 386 Cheonhodae-ro, Gwangjin-gu, Seoul 04808, Republic of Korea

⁵

Department of Regulatory Science, Graduated School, Kyung Hee University, Seoul 02447, Republic of Korea

⁶

Department of Fundamental Pharmaceutical Science, Graduated School, Kyung Hee University, Seoul 02447, Republic of Korea

^*

Authors to whom correspondence should be addressed.

Molecules 2024, 29(18), 4505; https://doi.org/10.3390/molecules29184505

Submission received: 26 August 2024 / Revised: 12 September 2024 / Accepted: 19 September 2024 / Published: 23 September 2024

(This article belongs to the Special Issue Advanced Analytical Tools for Characterization and Quality Control of Food, Drugs, and Natural Active Ingredients, 2nd Edition)

Download

Browse Figures

Versions Notes

Abstract

A highly accurate, precise, and simple liquid chromatography-tandem mass spectrometry (LC–MS/MS) method for ketotifen (KTF) estimation from Beagle dog plasma was developed and validated, with ketotifen-d3 (KTF-d3) as the internal standard (IS). KTF and IS were detected on an API 4000 mass spectrometer in multiple reaction monitoring (MRM) mode in electrospray ionization (ESI) positive ionization mode. The transitions were monitored at m/z 310.2 → 96.0 for KTF and m/z 313.2 → 99.1 for IS. KTF and IS were extracted from plasma using liquid-liquid extraction with methyl tertiary-butyl ether and then analyzed for 3 min with extracted samples (7 µL) into the LC–MS/MS system. Analytes were separated on a Luna^® Hilic column (50 × 2.0 mm i.d., 3 μm) using the Nexera X2 HPLC. The mobile phase A consisted of 10 mmol/L ammonium formate (pH 3.0), while mobile phase B consisted of 0.05% formic acid in acetonitrile. The ratio of mobile phase was 5:95 (v/v) at a flow rate of 0.2 mL/min. The method has been thoroughly validated in accordance with the bioanalytical method validation guidelines established by the Ministry of Food and Drug Safety in Korea and the U.S. Food and Drug Administration, addressing selectivity, lower limit of quantification, linearity, carryover, precision, accuracy, recovery, matrix effect, and stability. The developed LC–MS/MS method was effectively utilized for the bioequivalence assessment of ketotifen in Beagle dog plasma following the oral administration of ketotifen syrup.

Keywords: ketotifen; LC–MS/MS; Beagle dog plasma; syrup; bioanalytical method validation; bioequivalence

Share and Cite

MDPI and ACS Style

Song, E.; Shim, W.-S.; Choi, D.; Song, Y.; Jo, H.G.; Lee, S.; Jung, S.H.; Choi, Y.J.; Lee, K.-T. Development of a Simple and Validated LC–MS/MS Method for Quantitative Determination of Ketotifen in Beagle Dog Plasma and Its Application to Bioequivalence Study of Ketotifen Syrup Dosage Form. Molecules 2024, 29, 4505. https://doi.org/10.3390/molecules29184505

AMA Style

Song E, Shim W-S, Choi D, Song Y, Jo HG, Lee S, Jung SH, Choi YJ, Lee K-T. Development of a Simple and Validated LC–MS/MS Method for Quantitative Determination of Ketotifen in Beagle Dog Plasma and Its Application to Bioequivalence Study of Ketotifen Syrup Dosage Form. Molecules. 2024; 29(18):4505. https://doi.org/10.3390/molecules29184505

Chicago/Turabian Style

Song, Eunseo, Wang-Seob Shim, Doowon Choi, Yuna Song, Hyeong Geun Jo, Soobok Lee, Suk Han Jung, Yeo Jin Choi, and Kyung-Tae Lee. 2024. "Development of a Simple and Validated LC–MS/MS Method for Quantitative Determination of Ketotifen in Beagle Dog Plasma and Its Application to Bioequivalence Study of Ketotifen Syrup Dosage Form" Molecules 29, no. 18: 4505. https://doi.org/10.3390/molecules29184505

APA Style

Song, E., Shim, W.-S., Choi, D., Song, Y., Jo, H. G., Lee, S., Jung, S. H., Choi, Y. J., & Lee, K.-T. (2024). Development of a Simple and Validated LC–MS/MS Method for Quantitative Determination of Ketotifen in Beagle Dog Plasma and Its Application to Bioequivalence Study of Ketotifen Syrup Dosage Form. Molecules, 29(18), 4505. https://doi.org/10.3390/molecules29184505

Article Menu

Development of a Simple and Validated LC–MS/MS Method for Quantitative Determination of Ketotifen in Beagle Dog Plasma and Its Application to Bioequivalence Study of Ketotifen Syrup Dosage Form

Abstract

Share and Cite

Article Metrics

Article Access Statistics

Further Information

Guidelines

MDPI Initiatives

Follow MDPI