Enantioanalysis of Tertatolol in Plasma and Pharmaceutical Formulations with Immobilized Polysaccharide-Derived HPLC Chiral Column at Nano-detection Level

Abstract

A sensitive and selective high performance liquid chromatography (HPLC) method has been developed for the simultaneous determination of tertatolol enantiomers in plasma and pharmaceutical formulations. Enantiomeric resolution was achieved on cellulose tris(3,5-dichlorophenylcarbamate) immobilized onto 5 mm, spherical porous silica chiral stationary phase (CSP) known as Chiralpak IC with UV detection set at 254 nm. The mobile phase which consisted of n-hexane-ethanol-triethylamine (60:40:0.1%, v=v=v) was used with a flow rate of 0.3 mL=min. Bentazepam was chosen as internal standard to guarantee a high level of quantitative performance. The assay involved the use of solid phase extraction procedures for human plasma samples prior to HPLC analysis; C 18 cartridges gave good recoveries for both enantiomers without any interference. The stability of tertatolol enantiomers under different degrees of temperature was studied. The result shows that the drug was stable for at least 7 days at 70 C. The method was validated for its linearity, accuracy, precision, and robustness. The calibration curves in plasma were linear over the range of 25-1000 ng=mL for each enantiomer with a detection limit of 3 ng=mL. The mean relative standard deviation (RSD) of the results of within-day precision and accuracy of the drug were 2. There was no significant difference (P > 0.05) between inter-and intra-day studies for each enantiomer, which ensure the reproducibility of the assay method. The percentage recovery for both enantiomers from plasma was in the range of 91.3-98.7% at 75À800 ng=mL level for each enantiomer. The overall recoveries of tertatolol enantiomers from pharmaceutical formulation were in the range of 98-101% with %RSD ranged in 2.74-2.9%. The assay method is supposed to be suitable as therapeutic drug monitoring and chiral quality control for tertatolol formulations by HPLC.

Stereoselective HPLC analysis of tertatolol in rat plasma using macrocyclic antibiotic chiral stationary phase

“…The polar ionic mode (PIM) has been described as a novel mobile phase to obtain difficult enantioselective separation with macrocyclic antibiotic-based chiral stationary phases. 30 This approach uses a non-aqueous polar component (methanol) with glacial acetic acid or trifluoroacetic acid and an amine such as triethylamine which are necessary to achieve enantioseparation. In this study, baseline separation of the tertatolol enantiomers was achieved on the vancomycin CSP with a polar ionic mobile phase consisted of methanol-glacial acetic acid-triethylamine (100:0.01:0.015, v/ v/v) ( Table 1).…”

“…Recently, Hefnawy et al reported enantioanalysis of tertatolol in human plasma and pharmaceutical formulations using immobilized polysaccharide-derived HPLC chiral stationary phase known as Chiralpak IC column. 30 Reviewing the literature revealed that up to present time, no study has yet published concerning analysis of tertatolol enantiomers utilizing macrocyclic antibiotic CSPs.…”

Chiral Separations: Fundamental Review 2010

“…The detection of pindolol enantiomers in plasma and pharmaceutical products by HPLC with fluorescence detection was accomplished using a Chiralpak IB column . The Chiralpak IC CSP was used in the HPLC enantioanalysis of tertalolol enantiomers and also in the direct HPLC enantioseparation of terazosin .…”

Fases estacionárias quirais para cromatografia líquida de alta eficiência

“…Essas fases estacionárias apresentam excelente habilidade de reconhecimento quiral para uma série de compostos com diferentes propriedades físico-químicas. [78][79][80] Devido ao alto poder de enantiorresolução e grande estabilidade, as CSPs baseadas em derivados de polissacarídeos, adsorvidos ou quimicamente ligados ao suporte, têm sido utilizadas na quantificação de enantiômeros em matrizes biológicas [81][82][83][84] e para separações em escala semi-preparativa e preparativa.…”