Development and validation of a stereoselective HPLC method for the determination of the in vitro transport of nateglinide enantiomers in rat intestine

Abstract

A simple stereoselective high performance liquid chromatographic method was developed for the determination of the in vitro transport of the enantiomers of nateglinide (N-(trans-4-isopropylcyclohexyl-carbonyl)-phenylalanine) in the rat intestine using a Chiralcel OJ-RH column (150 x 4.0 mm, 5 microm). The effects of the mobile phase composition, pH, the flow rate, and the temperature on the chromatographic separation were investigated. The enantioseparation was achieved at 33 degrees C using a mobile phase containing 100 mM potassium dihydrogen phosphate, pH 2.5, and ACN (32:68 v/v) delivered at a flow rate of 1 mL/min. The analytes were monitored at 210 nm and linearity (r >0.99) was obtained for a concentration range of 0.5-50 microg/mL. The LOD and LOQ were 0.2 and 0.5 microg/mL for the R-enantiomer and 0.2 and 0.8 microg/mL for the S-enantiomer, respectively. Both, the intra- and interday accuracy and precision of the calibration curves were determined. The method was successfully applied to estimate the in vitro passage of the enantiomers and the racemate of nateglinide in duodenum, jejunum, and ileum of rats. Generally, higher concentrations of nateglinide and the S-enantiomer were observed when the racemate was administered compared to administration of the individual enantiomers of nateglinide.

Formulation, optimization, in vitro and in vivo evaluation of Nateglinide-loaded nanostructured lipid carriers for enhanced bioavailability.

“…Furthermore the relative bioavailability (F) of NTG-NLC was assessed by establishing the ratio between the AUC of NTG-NLC and AUC of NTG suspension. [41,42] Relative Bioavailability (F)…”

Determination of the L-enantiomer of nateglinide in pharmaceutical formulations by micellar electrokinetic chromatography

“…Another paper showed that enantioseparation of nateglinide and its enantiomer was achieved at ambient temperature on a Chiralcel OD-R column (Qi et al, 2003). The in vitro transport of the enantiomers of nateglinide in the rat intestine was determined by chiral HPLC using a Chiracel OJ-RH column (Maddi et al, 2007). Molecularly imprinted polymers (MIP) and monolithic moleculary imprinted polymers were used as chiral stationary phase for the determination of nateglinide and its L-enantiomer (Yang et al, 2004;Yin et al, 2005).…”

Atropisomeric Separation of PCB-95 by HPLC

“…Chromatographic separation of enantiomers can be achieved either by forming diastereoisomers by chiral derivatization agents or by using a chiral stationary phase [27]. The use of chiral derivatization agent requires a functional group in the analyte such as an amine, hydroxyl, carboxyl, carbonyl etc.…”

“…Among them, cellulose, amylose ester and carbamate polymer derivatives coated onto a large pore silica backbone have proven to be most successful for chiral resolution using a range of mobile phases from polar to non-polar [28; 32-33]. Commercial columns containing these CSPs include Chiralcel OD [cellulose tri (3 [27,30].…”