HPLC‐Fluorescence Method for the Enantioselective Analysis of Propranolol in Rat Serum Using Immobilized Polysaccharide‐Based Chiral Stationary Phase

Abstract

A stereoselective high-performance liquid chromatographic (HPLC) method was developed and validated to determine S-(-)- and R-(+)-propranolol in rat serum. Enantiomeric resolution was achieved on cellulose tris(3,5-dimethylphenylcarbamate) immobilized onto spherical porous silica chiral stationary phase (CSP) known as Chiralpak IB. A simple analytical method was validated using a mobile phase consisted of n-hexane-ethanol-triethylamine (95:5:0.4%, v/v/v) at a flow rate of 0.6 mL min(-1) and fluorescence detection set at excitation/emission wavelengths 290/375 nm. The calibration curves were linear over the range of 10-400 ng mL(-1) (R = 0.999) for each enantiomer with a detection limit of 3 ng mL(-1). The proposed method was validated in compliance with ICH guidelines in terms of linearity, accuracy, precision, limits of detection and quantitation, and other aspects of analytical validation. Actual quantification could be made for propranolol isomers in serum obtained from rats that had been intraperitoneally (i.p.) administered a single dose of the drug. The proposed method established in this study is simple and sensitive enough to be adopted in the fields of clinical and forensic toxicology. Molecular modeling studies including energy minimization and docking studies were first performed to illustrate the mechanism by which the active enantiomer binds to the β-adrenergic receptor and second to find a suitable interpretation of how both enantiomers are interacting with cellulose tris(3,5-dimethylphenylcarbamate) CSP during the process of resolution. The latter interaction was demonstrated by calculating the binding affinities and interaction distances between propranolol enantiomers and chiral selector.

Quantitative Analysis of Drugs in Biological Matrices by HPLC Hyphenated to Fluorescence Detection

“…The LOQ was 17 ng/ml for each isomer. Recently Alazani et al [85] reported an enantioselective method to quantify propranolol in rat serum. Propranolol is a β-adrenergic antagonist administered as a racemic mixture in the treatment of hypertension and several other diseases.…”

State-of-the-art and recent developments of immobilized polysaccharide-based chiral stationary phases for enantioseparations by high-performance liquid chromatography (2013–2017)

“…have been determinated in environmental or food samples (soil, natural waters, fruits, vegetables, etc.). Pharmaceutical compounds recognized as potential anti-carcinogen agents [44,45,56,92,64], α-and β-adrenergic receptor blockers [51,92,59,[65][66][67][68]60], nonsteroidal anti-inflammatory drugs [38], antihistamines [92,74], proton pump inhibitors [50,94,73,79,81,88,96,76], retroviral drugs [55,84], antihyperglycemic drugs [83], antihelminthic agents [40,57], anti-hypertensive drugs [41,53,85], L-type calcium channel [72,89], broad-spectrum antimicrobials [95,70], nootropic drugs [90,91], anticoagulant drugs [69], etc. have also been determined, especially in commercial products or biological samples (pharmaceutical formulations, human urine, human or animal blood, human plasma, animal serum, etc.).…”

“…Differences in the formation of hydrogen bonds between the enantiomers of propranolol and cellulose tris-(3,5-dimethylphenylcarbamate) derived from molecular modeling were used to explain the elution order observed in HPLC [66]. The less retained (S)-enantiomer formed a hydrogen bond between the hydroxy group of the selectand and the carbamate carbonyl group of the selector while the stronger retained (R)enantiomer formed hydrogen bonds with the carbohydrate backbone due to deeper insertion into the chiral cavities.…”

Synthesis and Enantioseparation Ability of Xylan Bisphenylcarbamate Derivatives as Chiral Stationary Phases in HPLC

“…As the key of these techniques, the design and preparation of efficient chiral stationary phases (CSPs) continue attracting much attention and a large number of CSPs with various structures have been developed for direct enantioseparation of racemic compounds for diverse purposes. Those based on the phenylcarbamate and benzoate derivatives of cellulose and amylose have proved to be versatile CSPs for HPLC and SFC, and a wide range of enantiomers have been successfully separated on them …”

“…Those based on the phenylcarbamate and benzoate derivatives of cellulose and amylose have proved to be versatile CSPs for HPLC and SFC, and a wide range of enantiomers have been successfully separated on them. [3][4][5][6][7][8][9][10][11][12][13] Besides cellulose and amylose, a series of phenylcarbamate derivatives of other polysaccharides including chitin, chitosan, xylan, dextran, and so on, have also been prepared, and their chiral resolving powers were previously evaluated. 14,15 Among these derivatives, the 3,5-dimethylphenylcarbamates of chitosan and xylan exhibited relatively higher recognition ability for some chiral compounds when compared to those based on cellulose and amylose, indicating the potential to develop the new CSPs with a high chiral recognition ability based on these two polysaccharides.…”