Derivatization of Methylglyoxal for LC-ESI-MS Analysis—Stability and Relative Sensitivity of Different Derivatives

The great research interest in the quantification of reactive carbonyl compounds (RCCs), such as methylglyoxal (MGO) in biological and environmental samples, is reflected by the fact that several publications have described specific strategies to perform this task. Thus, many reagents have also been reported for the derivatization of RCCs to effectively detect and quantify the resulting compounds using sensitive techniques such as liquid chromatography coupled with mass spectrometry (LC-MS). However, the choice of the derivatization protocol is not always clear, and a comparative evaluation is not feasible because detection limits from separate reports and determined with different instruments are hardly comparable. Consequently, for a systematic comparison, we tested 21 agents in one experimental setup for derivatization of RCCs prior to LC-MS analysis. This consisted of seven commonly employed reagents and 14 similar reagents, three of which were designed and synthesized by us. All reagents were probed for analytical responsiveness of the derivatives and stability of the reaction mixtures. The results showed that derivatives of 4-methoxyphenylenediamine and 3-methoxyphenylhydrazine—reported here for the first time for derivatization of RCCs—provided a particularly high responsiveness with ESI-MS detection. We applied the protocol to investigate MGO contamination of laboratory water and show successful quantification in a lipoxidation experiment. In summary, our results provide valuable information for scientists in establishing accurate analysis of RCCs

Analytical Method Development and Validation of Stability Indicating RP-HPLC Method for The Simultaneous Estimation of Dolutegravir, Emtricitabine and Tenofovir Disproxil Fumarate

A simple, Accurate, precise method was developed for the simultaneous estimation of the Dolutegravir (DUA), Emtricitabine(ECB) and Tenofovir Disoproxil fumarate (TDF) tablet dosage form. Chromatogram was run through Std Kinetex Biphenyl, 250 x 4.6 mm, 5µm. Mobile phase-A containing pH-3.0 Ammonium acetate Buffer & Mobile phase -B containing Degassed mixture of Acetonitrile & Methanol taken in the ratio of 50:50 was pumped through column at a flow rate of 1 mL/min. Diluent used in this method was 0.1% Orthophosphoric acid buffer & Methanol in the ratio of 50:50%v/v. Temperature was maintained at 40°C. Optimized wavelength selected was 260 nm. Retention time of DUA, ECB and TDF were found to be 14.6min, 3.98min and 12.60min. %RSD of the DUA, ECB and TDF were found to be 0.35, 0.47 & 0.40 respectively. %Recovery was obtained as 98.9%, 99.6% and 99.3% for DUA, ECB and TDF respectively. Both the runtime and retention times were decreased, so this method developed was simple and economical this can be adopted in regular quality control in pharma industries