Trace determination of 1-octacosanol in rat plasma by solid-phase extraction with Tenax GC and capillary gas chromatography.

1-Octacosanol is the major component of policosanol, a new natural lowering-cholesterol agent. A sensitive solid-phase extraction with a Tenax GC capillary gas chromatography method for determining the proportion of this fatty alcohol in plasma after its denaturation with trichloroacetic acid was developed. The trimethylsilyl ether derivative of the target analyte obtained from the organic extract showed excellent chromatographic properties and was detectable in the low nanogram range (1 ng/ml). Adequate separation from plasma's extract was achieved with a fused-silica capillary column (30m x 0.25 mm I.D.) with SPB-5 (0.5 microm film thickness) and operated with temperature programming from 100 to 200 degrees C at 40 degrees C/min and from 200 degrees C increased at 10 degrees C/min to 320 degrees C, then held for 30 min, the carrier gas flow-rate (argon) was 1 ml/min. Quantification was performed by the internal standard method using 1-hexacosanol. The reliable relative retention parameters and the mass response factors values, and their confidence levels, ensure a proper GC sensitivity, necessary for the determination of the alcohol being analyzed. The method was evaluated to a concentration range from 6 to 47.6 ng/ml of plasma obtaining recoveries from 95 to 98%. The correlation between the theoretical concentration values and the corresponding experimental values was appropriate (gamma=0.9718 x -0.0915; r2=0.9998). The method showed a good within-day (RSD=4.3%) and between-day (RSD=6.0%) precision according to the acceptance criteria (<10%). This procedure was successfully applied to the study of 1-octacosanol in rat plasma samples after a single oral administration (40 mg/kg) of policosanol.

Group method approach to the estimation of response factors of unavailable substances in quantitative gas chromatography.

The method's accuracy of a compound quantitation by chromatography depends on the calibration procedure with a pure standard of the target analyte, if the latter is unavailable uncertainty is unavoidable. The group method is a different approach in GC quantitative analysis that shows a practicable way for avoiding this uncertainty and accurately quantify a mixture containing one or more unavailable components. This paper is concerned with the definition of the group method quantitative parameters, the application procedures for their calculation, the determination of the quantitative proportion of a group of unavailable components of a mixture and the partial or total quantitation of the latter. The paper also describes the steps for carrying out the so-called group-correlation method in the determination of the response factors of unavailable compounds, which belong to a homologous series. The GC experimental corroboration of the group method approach employing model mixtures of compounds is also presented.

Analytical procedure for the determination of 1-octacosanol in plasma by solvent extraction and capillary gas chromatography.

A simple and rapid method for the determination of 1-octacosanol in plasma, based on an improved Folch extraction technique and capillary gas chromatography, has been developed taking into account the analytical criteria for the pharmacokinetic studies. The procedure was validated in the range of 50-2000 ng/ml. Despite the complexity of the obtained fingerprints, the efficiency and the separation power of GC allowed the determination of 1-octacosanol in plasma samples. The high recoveries (94.5-98.7%) and precision (1.8-5.8%) obtained are in accordance with the established validation criteria. The validity of this method for pharmacokinetic purposes was shown using an endovenous experiment in animals.