Simultaneous HPLC-UV determination of ketamine, xylazine, and midazolam in canine plasma.

An isocratic reversed-phase high-performance liquid chromatography method with UV detection is developed and validated for the simultaneous determination of ketamine, xylazine, and midazolam in canine plasma. Analytes are extracted from alkalinized samples into diethyl ether-methylene chloride (7:3, v:v) using single-step liquid-liquid extraction. Chromatographic separation is performed on a C(18) column using a mobile phase containing an acetonitrile-methanol-10 mM sodium heptanesulfonate buffer adjusted to pH 3, with glacial acetic acid (44:10:46, v:v) at a detection wavelength of 210 nm, with a total runtime of 10 min. The calibration is linear over the range of 78.125-5000 ng/mL for ketamine and 15.625-1000 ng/mL for xylazine and midazolam. The limits of detection are 17.8, 10.3, and 15.1 ng/mL for ketamine, xylazine, and midazolam, respectively. The extraction recoveries are 76.1% for ketamine, 91.0% for midazolam, and 78.2% for xylazine. The method is successfully used for clinical and pharmacokinetic studies of the three-drug fixed dose combination formulations.

Determination of trace trichlorfon by high performance liquid chromatography with UV detection based on its catalytic effect on sodium perborate oxidizing benzidine.

Trichlorfon has the capacity to catalyze the oxidation of benzidine (4,4'-diamino-biphenyl) to 4-amino-4'-nitro biphenyl in the presence of sodium perborate. The product of the catalyzed reaction was validated by LC-MS method. Reversed-phase high performance liquid chromatography with 365 nm UV detection was used for separation and quantification of 4-amino-4'-nitro biphenyl. It can be proven there is a linear relationship between the peak areas of 4-amino-4'-nitro biphenyl and trichlorfon in the concentration range of 0.02-0.5 mg L(-1) (r=0.9988). Limit of detection was 2.0 microg L(-1). A method for the indirect determination of trichlorfon using HPLC was developed based on catalytic effect of trichlorfon. Method validation was performed on samples spiked at three levels (0.5, 1.0, 1.5 mg kg(-1)), the recoveries ranged from 67.5 to 82.1%, with relative standard deviations between 4.5 and 7.3%. 0.01 mol L(-1) sodium dodecyl sulphate (SDS) solution was used to extract trichlorfon from samples and solid-phase extraction was used to isolate and concentrate trichlorfon in SDS solution. The recoveries of trichlorfon obtained with percolating the extraction through a SPE system were essentially in agreement with those obtained by liquid-liquid extraction. This new isolation technique decreases the use of toxic solvents and satisfies the requirements of Green Analytical Chemistry.

[Analysis of cis-9, trans-11-conjugated linoleic acid in milk fat by capillary gas chromatography].

Conjugated linoleic acid (CLA) is a term representing a mixture of positional and geometric isomers of octadecadienoic acid with a conjugated double bond system. Conjugated linoleic acid has attracted a great deal of interest among nutritionists because it is a natural fat component that appears to have a number of health improvement properties. The cis-9, trans-11-CLA is the major CLA isomer found in dairy products accounting for 75% to 90% of the total CLA in milk fat. A capillary gas chromatographic method equipped with a flame ionization detector for the analysis of the cis-9, trans-11-CLA in milk fat was developed. The cis-9, trans-11-CLA was extracted with hexane-isopropanol, methylated with methanol-sodium methylate and cis-9, trans-11-CLA was separated and quantified using gas chromatography. Retention time of the peaks was used for qualitative analysis, while external standard method was used for quantitative analysis. The recovery of the cis-9, trans-11-CLA was 100.26%. The relative standard deviation was 1.9% (n = 6). This method presented is advantageous for high precision, high sensitivity analysis with smaller sample size and simpler pretreatment. It would be of significance for analyzing the contents of other fatty acids in the milk and milk products.