Analysis of corticosterone in rat plasma by high-performance liquid chromatography
| Autor: | Chien Bm, Wong Yn, D'mello Ap |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
Detection limit
Male Chromatography Coefficient of variation Extraction (chemistry) Reproducibility of Results General Chemistry High-performance liquid chromatography Circadian Rhythm Rats Standard curve Rats Sprague-Dawley chemistry.chemical_compound chemistry Corticosterone Sodium hydroxide Stress Physiological Blood plasma Animals Spectrophotometry Ultraviolet Chromatography High Pressure Liquid |
| Zdroj: | Journal of chromatography. B, Biomedical applications. 661(2) |
| ISSN: | 1572-6495 |
| Popis: | A sensitive and specific high-performance liquid chromatographic assay for the determination of corticosterone in rat plasma using dexamethasone as the internal standard is reported. Rat plasma (0.5 ml) is extracted with methylene chloride, washed with 0.1 M sodium hydroxide and then with water. The extract is analyzed by HPLC on a C 18 column with ultraviolet absorbance detection at 254 nm. Pooled rat plasma was treated with activated decolorizing carbon to remove endogenous corticosterone, and was then used to prepare standards for the assay. Using 0.5 ml plasma for extraction, the detection limit of the assay is 10 ng/ml. The standard curve is linear over the concentration range 10–500 ng/ml. The recovery of corticosterone after extraction was independent of concentration and ranged from 87 to 95%. The coefficient of variation for intra-day and inter-day precision ranged from 2.4 to 7.4% and 2.1 to 8.7%, respectively. In addition, for concentrations ranging from 10 to 500 ng/ml the accuracy is within 5% of the spiked standards. The assay was utilized to examine the circadian rhythm of plasma corticosterone, and to examine the effect of immobilization stress on corticosterone levels in rats. |
| Databáze: | OpenAIRE |
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