Home | Articles
Published on:October 2020
Indian Journal of Pharmaceutical Education and Research, 2020; 54(3s):s671-s676
Original Article | doi:10.5530/ijper.54.3s.167

Method Development and Validation of Ephedrine and Pseudoephedrine in Horse Urine by Liquid Chromatography-Tandem Mass Spectrometry


Authors and affiliation (s):

Ashok Kumar Maurya1, Puran Lal Sahu1, Sakshi Jethi1, Himank Gosain2, Kapendra Sahu1,*

1National Dope Testing Laboratory, Ministry of Youth Affairs and Sports (MYAS), Government of India, New Delhi, INDIA.

2Department of Chemistry, Manav Rachna University, Haryana, INDIA.

Abstract:

Background: A comprehensive, fast and precise method for the quantification of ephedrine and pseudoephedrine in horse urine was established and validated utilizing Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) followed by the necessities of Association of Official Racing Chemists (AORC), Federation of Equine International (FEI) and International Council for Harmonization (ICH) guidelines. Methodology: The processing of samples was performed by de conjugation accompanied by enzymatic hydrolysis, Solid Phase Extraction (SPE) procedure using HLB cartridges. Results: The total chromatographic run time of this method was 7.0 min and found linear over the concentration range of 4-40 ng/mL with a correlation of coefficient (r2) of 0.99. The intraday and inter-day assay precision of ephedrine were extended from 1.64 to 2.86% and 1.61 to 3.12 %, respectively. In context of pseudoephedrine, the intraday and interday assay precision were ranged from 1.55 to 3.26% and 1.11 to 2.60 %, respectively. The absolute percent recovery (%) was found to be 85% for Ephedrine and 90% for Pseudoephedrine. The percent recovery was adequate to discriminate and quantitate the Ephedrine and Pseudoephedrine at or below the level prescribed by AORC Proficiency Testing (PT) list ie 20ng/ ml for equine urine sample. The LOD and LOQ were found 2 and 4 ng/ml respectively for Ephedrine and Pseudoephedrine. Conclusion: A comprehensive and precise method for quantitative estimation and diastereomeric differentiation of Ephedrine and Pseudoephedrine by LC-MS/MS was well established and validated according to AORC, FEI and ICH guidelines. This strategy may be additionally utilized for remedial medication checking purposes.

Key words: Ephedrine, Pseudoephedrine, ICH Guidelines, Doping control, Mass spectrometry, AORC, Validation.

 




 

Impact Factor

IJPER - An Official Publication of Association of Pharmaceutical Teachers of India is pleased to announce continued growth in the Latest Release of Journal Citation Reports (source: Web of Science Data).

 

Impact Factor® as reported in the 2023 Journal Citation Reports® (Clarivate Analytics, 2023): 0.8

The Official Journal of Association of Pharmaceutical Teachers of India (APTI)
(Registered under Registration of Societies Act XXI of 1860 No. 122 of 1966-1967, Lucknow)

Indian Journal of Pharmaceutical Education and Research (IJPER) [ISSN-0019-5464] is the official journal of Association of Pharmaceutical Teachers of India (APTI) and is being published since 1967.

DOI HISTORY

IJPER uses reference linking service using Digital Object Identifiers (DOI) by Crossref. Articles from the year 2013 are being assigned DOIs for its permanent URLs