Analysis of Benzodiazepines in Plasma Samples by Dllme and Lc-Dad: Critical Aspects, Flaws and Issues Encountered – A Discussion

Abstract

Dispersive liquid-liquid microextraction (DLLME) is a fast and simple technique and applicable procedure for the analysis of xenobiotics in different biological matrices. We have developed a DLLME method for the determination of benzodiazepines in plasma samples using liquid chromatography-diode-array detection (LC-DAD). During method validation, relative standard deviation values of more than 20% were observed for both intra and inter-day associated with precision and accuracy. So, full method validation was unsuccessful. Critical aspects and flaws associated with DLLME for benzodiazepines determination in biological matrices has never been addressed. The majority of previous studies involving DLLME for benzodiazepines analysis either did not carry out a full method validation or they do not share any details regarding their full validation procedure. The current study has focused on the issues and pitfalls encountered upon method validation so that scientific groups worldwide can avoid unnecessary experiments and benefit from the present discussion. These observations reinforce the need to carry out such a process to guarantee the reliability of the results, even more so if one is intending its application for real case samples in the field of toxicological analysis. Indeed, full, and satisfactory method validation is paramount to provide adequate and trustworthy analytical results.