Development and Validation of High-Performance Liquid Chromatography–Diode Array Detector Method for the Determination of Tramadol in Human Saliva

Abdel- Aziz,Y El- Sayed,Maha A.Hilal,Khaled M Mohamed

doi:10.4172/2157-7064.1000114

Abstract

A high-performance liquid chromatography–diode array detector method (HPLC-DAD) was developed and validated for the determination of tramadol in human saliva. Samples of saliva were prepared utilizing liquid–liquid extraction with hexane-ethyl acetate (4:1, v/v). Propranolol was used as an internal standard (IS). The linear range and average recovery of tramadol were 0.25–4.00 μg/mL and 94.70%, respectively. The intra- and inter-assay precisions at low, intermediate and high concentrations were within 1.40–9.79 % and 0.77–3.98%, respectively. Intra- and inter-assay accuracies were within 98.50–107.13% and 100.44–103.89%, respectively. The proposed method was successfully applied to determine tramadol in saliva concentrations in three healthy volunteers for 24 hours after administration of 100 mg oral doses of tramadol.

Highlights

Screening and quantitative analysis of prescription medications as well as the more common drugs of abuse is part of any systematic toxicological analysis
The methods described for the determination of tramadol in biological samples involve gas chromatography (GC) with nitrogen–phosphorous detection [6], flame ionization detection [7] or mass spectrometry (MS) [4,8,9,10,11,12,13,14,15,16,17]
The dried extracts were reconstituted in 50 μL acetonitrile, vortex mixed for 30 seconds and 20 μL was injected into the HPLC system

Summary

Introduction

Screening and quantitative analysis of prescription medications as well as the more common drugs of abuse is part of any systematic toxicological analysis. One of the major techniques used at present for such screening are HPLC-DAD [1,2,3]. While blood and urine are more commonly used for these test profiles, oral fluid (saliva) is increasing in popularity as an alternative matrix, due to its ease of collection, difficulty of adulteration and improving sensitivity of analytical techniques. This is due to saliva is less complex than blood, with fewer proteins, characteristics suggesting the possibility of successful measurement with limited sample preparation, such as dilution, protein precipitation, or centrifugation and direct injection [4]. The aim of the present work is to develop simple and sensitive analytical method for the determination of tramadol in saliva

Objectives

Methods

Results