betadex and octadecylsilane

Non-porous C18 silica gel stationary phase (1 µm) was prepared and applied to chiral separation in pressurized capillary electrochromatography (pCEC) for the enantioseparation of various basic compounds. The non-porous silica particles (1 µm) were synthesized using modified St6ber method. C18 stationary phase (1 µm) was prepared by immobilization of chloro-dimethyl-octadecylsilane. Using carboxymethyl-β-cyclodextrin (CM-β-CD) as the chiral additive, the pCEC conditions including the content of acetonitrile (ACN), concentration of buffer, pH, the concentration of chiral additive and flow rate as well as applied voltage were investigated to obtain the optimal pCEC conditions for the separation of four basic chiral compounds. The column provided an efficiency of up to 190,000 plates/m. Bupropion hydrochloride, clenbuterol hydrochloride, metoprolol tartrate, and esmolol hydrochloride were baseline separated under the conditions of 5 mmol/L ammonium acetate buffer at pH 4. 0 with 20% (v/ v) acetonitrile, and 15 mmol/L CM-β-CD as the chiral additive. The applied voltage was 2 kV and flow rate was 0.03 mL/min with splitting ratio of 300:1. The resolution were 1.55, 2.82, 1. 69, 1. 70 for bupropion hydrochloride, clenbuterol hydrochloride, metoprolol tartrate, esmolol hydrochloride, respectively. The C18 coverage was improved by repeating silylation method. The synthesized 1 µm C18 packings have better mechanical strength and longer service life because of the special, non-porous structure. The column used in pCEC mode showed better separation of the racemates and a higher rate compared with those used in the capillary liquid chromatography (cLC) mode. This study provided an alternative way for the method of pCEC enantioseparation with chiral additives in the mobile phase and demonstrated the feasibility of micron particle stationary phase in chiral separation.

Topics: beta-Cyclodextrins; Bupropion; Capillary Electrochromatography; Chromatography, Liquid; Clenbuterol; Metoprolol; Propanolamines; Silanes; Silica Gel; Silicon Dioxide

A sensitive and stereospecific high-performance liquid chromatography (HPLC) method for the quantitation of ketoprofen enantiomers in human serum was developed. The assay involves the use of an octadecylsilane solid-phase extraction for serum sample clean-up prior to HPLC analysis. Chromatographic resolution of the ketoprofen enantiomers was performed on a nonporous octyldecylsilane column with hydroxypropyl beta-cyclodextrin as the mobile phase additive. The composition of the mobile phase was 98:2 v/v aqueous 0.1% trifluoroacetic acid (TFA), pH 4.00 (adjusted with triethylamine (TEA))/acetonitrile containing 10 mM hydroxypropyl beta-cyclodextrin (beta-CD) at a flow rate of 0.8 ml min-1. Recoveries of R(-)-ketoprofen was 95.4 +/- 2.16% and for S(+)-ketoprofen 96.2 +/- 1.31%. Linear calibration curves were obtained in the range 0.025-15 micrograms ml-1 range for each enantiomer in serum. The detection limit based on a S/N = 3 ratio was 10 ng ml-1 for each enantiomer in serum with ultraviolet detection at 220 nm. The limit of quantitation for each enantiomer was 25 ng ml-1. Precision calculated as % relative standard deviation (%R.S.D.) and accuracy calculated as % error were in the range 0.2-5.2% and 0.3-2.2%, respectively, for the R enantiomer and 0.3-6.2 and 0.2-3.2%, respectively, for the S enantiomer.

Topics: 2-Hydroxypropyl-beta-cyclodextrin; Anti-Inflammatory Agents, Non-Steroidal; beta-Cyclodextrins; Chromatography, High Pressure Liquid; Cyclodextrins; Humans; Indicators and Reagents; Ketoprofen; Reproducibility of Results; Sensitivity and Specificity; Silanes; Stereoisomerism