A rapid, sensitive, and reproducible pre-column derivatisation procedure has been established for the simultaneous determination of 20 amino acids by high-performance liquid chromatography using fluorescence detection. The amino acids were derivatized using o-phthalaldehyde and 9-fluorenylmethyl-chloroformate reagents. The optimal conditions for simultaneous separation and detection of both primary and secondary amino acids were investigated. The developed method has several advantages, namely automated pre-column derivatization, short analysis time with optimal separation, a simple and economical mobile phase, high level of precision for peak area and retention time, and higher sensitivity with more reliability of peak identification. The biological media development is the key parameter for macromolecule drug discovery. Biological media amino acids in three consecutive discovery batches were determined and the results showed a good agreement with hypothetical value. The method appears suitable for application to measure biological media amino acids at various stages of macromolecule drug discovery.
β-blockers are a class of antihypertensive drugs that are used for the management of cardiac arrhythmias, cardioprotection after myocardial infarction (heart attack) and hypertension. They have revolutionized the medical management of angina pectoris and are recommended as first-line agents by national and international guidelines. Although β-blockers are still the cornerstone for the treatment of heart failure, some of the drugs in this category are prohibited in several sports requiring vehicle control and bodily movements as they reduce heart rate and tremors, and improve performance. As a result, urine analysis of β-blockers is mandatory in doping control and toxicological screening. The determination of plasma levels of β-blockers helps to ensure noncompliance in patients with persistent hypertonia to confirm the diagnosis of β-blocker poisoning and for therapeutic drug monitoring. This review provides a comprehensive account of various analytical methods developed for detection and quantitation of β-blockers in plasma and urine.
Inclusion complex formation of atenolol with -cyclodextrin (-CD) has been investigated by HPLC on different stationary phases, by varying pH and concentration of -CD added as an additive in the mobile phase over a wide range of column temperature. Stationary phases of different polarity and hydrophobicity were evaluated to find the best conditions for complex formation. The optimum conditions for inclusion complexation were achieved on YMC ODS-AQ C18 ( mm, 5 μ) analytical column. The apparent formation constant () of the complex as evaluated by liquid chromatography using retention factors () was M−1 at 25°C. The stoichiometry of the complex was 1 : 1 as is evident from the straight line plot of 1/ versus -CD concentration. The formation of inclusion complex was essentially enthalpy (−42.12 kJ/mol) driven and the binding forces included hydrophobic, van der Waals-London dispersion interactions. The enthalpy-entropy compensation criterion was used to prove the inclusion phenomena.
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