Quantification of ondansetron, granisetron and tropisetron in goat plasma using hydrophilic interaction liquid chromatography-solid phase extraction coupled with hydrophilic interaction liquid chromatography-triple quadrupole tandem mass spectrometry.

An assay method to quantify ondansetron (OND), granisetron (GRA) and tropisetron (TRO) in goat plasma has been successfully developed and validated. This method procedure for the analysis of OND, GRA and TRO was involved of extracting samples with hydrophilic interaction liquid chromatography (HILIC) solid phase extraction (SPE) and determination by liquid chromatography coupled to tandem mass spectroscopy. An SPE method for the simultaneous extraction of OND, GRA and TRO with high efficiency and selectivity was developed. Prior to HPLC-MS/MS analysis, most of the sources of interference present in the supernatant after protein precipitation of plasma proteins was efficiently removed from the samples by the HILIC SPE treatment. For the quantification of OND, GRA and TRO in the samples, tandem mass spectrometry operating in positive electrospray ionization mode with multiple reaction monitoring was used. The calibration curve was performed in the range of 0.2-20 ng/mL for the target OND, GRA and TRO in goat plasma samples. The precision of the intra- and inter-day assay for OND, GRA and TRO were 1.84-6.23% and 3.89-5.31%, 2.63-6.29% and 3.76-5.31%, 1.99-5.67% and 2.64-4.70%, respectively. The accuracy of the intra- and inter-day assay for OND, GRA and TRO were 89.15-97.39% and 89.46-95.17%, 91.08-100.82% and 91.24-99.47%, 92.30-100.74% and 94.21-97.90%, respectively. For the determination of OND, GRA and TRO in plasma samples, no significant matrix effects were observed. The mean absolute recoveries were 103-150%, 115-121%, and 98-141% for OND, GRA and TRO, respectively. Furthermore, the mean process efficiency values of silica SPE were 98-135%, 92-124%, and 72-109% for OND, GRA and TRO, respectively.

Separation behavior of basic compounds on unbonded silicon oxynitride and silica high-performance liquid chromatography stationary phases with reversed-phase eluents.

Unbonded silicon oxynitride and silica high-performance liquid chromatography stationary phases have been evaluated and compared for the separation of basic compounds of differing molecular weight, pKa , and log D using aqueous/organic mobile phases. The influences of percentage of organic modifier, buffer pH, and concentration in the mobile phase on base retention were investigated on unbonded silicon oxynitride and silica phases. The results confirmed that unbonded silicon oxynitride and silica phases demonstrated excellent separation performance for model basic compounds and both the unbonded phases examined possessed a hydrophobic/adsorption and ion-exchange character. The silicon oxynitride stationary phase exhibited high hydrophilicity compared with silica with a reversed-phase mobile phase. An ion-exclusion-type mechanism becomes predominant for the separation of three aimed bases on the silicon oxynitride column at pH 2.8. Different from silicon oxynitride stationary phase, no obvious change for the retention time of three model bases on silica stationary phase at pH 2.8 can be observed.

Evaluation of a silicon oxynitride hydrophilic interaction liquid chromatography column in saccharide and glycoside separations.

The retention characteristics of a silicon oxynitride stationary phase for carbohydrate separation were studied in hydrophilic interaction chromatography mode. Four saccharides including mono-, di-, and trisaccharides were employed to investigate the effects of water content and buffer concentration in the mobile phase on hydrophilic interaction liquid chromatography retention. For the tested saccharides, the silicon oxynitride column demonstrated excellent performance in terms of separation efficiency, hydrophilicity, and interesting separation selectivity for carbohydrates compared to the bare silica stationary phase. Finally, the silicon oxynitride hydrophilic interaction liquid chromatography column was employed in the separation of complex samples of fructooligosaccharides, saponins, and steviol glycoside from natural products. The resulting chromatograms demonstrated good separation efficiency and longer retention compared with silica, which further confirmed the advantages and potential application of silicon oxynitride stationary phase for hydrophilic interaction liquid chromatography separation.

[Synthesis of porous spherical silicon oxynitride material and evaluation of its properties in reversed-phase chromatographic separation].

Silica has been widely used as HPLC column packing material. However, the fact that base can attack the silanol and dissolve the silica embarrasses the utilization of silica stationary phase in high pH mobile phases (pH >8). In our previous research, the use of porous spherical silicon oxynitride (sph-SiON) material from high temperature nitridation of silica microspheres as stationary phase for HPLC has been explored, and the sph-SiON is stable to alkaline mobile phases and demonstrates excellent separation of a variety of polar compounds in hydrophilic interaction liquid chromatography (HILIC) mode. Herein, the degree of nitridation was studied as a function of temperature of nitridation at 750-1 050 degrees C, yielding the silicon oxynitride with 0.40%-12.0% (mass fraction) nitrogen from elemental analysis. At the temperature of 1 050 degrees C, the nitrogen content increased from 12.0% to 24.5% with the nitridation time increasing from 20 h to 120 h. The sph-SiON is stable when disposed in different pH aqueous solutions for one week. The sph-SiON material can be modified to give hydrophobic surface through the reaction of surface Si-NHx with dimethyloctadecylchlorosilane. Elemental analysis and 13C cross-polarization magic-angle spinning (CP/MAS) NMR spectrum of C18-sph-SiON prove the integration of C18 alkyl groups attached onto the sph-SiON surface. The chromatographic evaluation of C18-sph-SiON in reversed-phase separation mode was performed with alkylbenzenes as hydrophobic probes. Three alkylbenzene compounds can be separated and retained well on C18-sph-SiON even in the mobile phase of methanol/H2O (70/30, v/v) with 78 507 plates/m, and an excellent tailing factor (0.95) can be obtained for ethylbenzene. In comparison with C18-SiO2, C18-sph-SiON shows distinct differences with respect to different classes of analytes, i. e. neutral analyte naphthalene, acidic analyte ibuprofen, and basic analyte amitriptyline.

[Determination of Huperzine A in the extract of Huperzia serrata by high performance liquid chromatography].

A high performance liquid chromatographic (HPLC) method was established to determine the Huperzine A in the extract of Huperzia serrata. After extracted by methanol/ water/formic acid (10/90/0.2, v/v/v), the sample was filtered for HPLC analysis. The separation was performed on an XCharge C18 column (150 mm x 4.6 mm, 5 microm) by gradient elution with water (containing 0.1% (v/v) trifluoroacetic acid) and acetonitrile (containing 0.09% (v/v) trifluoroacetic acid) as the mobile phases. Rapid separation was achieved within 10 min at a flow rate of 2 mL/min with ultraviolet absorption detection at a wavelength of 310 nm. Under the optimized conditions, good linearity was obtained in the range of 2. 12 - 106 mg/L with the correlation coefficient (R2) of about 0.999 9. The average recovery was 102.34% with the relative standard deviation (RSD) of 0.46%. The intraday and interday precisions were all below 2%. The results demonstrate that this method is simple, rapid and accurate with good reproducibility, and can be used to evaluate the quality of Huperzia serrata.