Determination of physostigmine in plasma by high-performance liquid chromatography and fluorescence detection
| Autor: | Suellen Ferraris, John R Ryabik, Don W. Korte, Conrad R. Wheeler, Nabil M. Elsayed |
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| Rok vydání: | 1989 |
| Předmět: |
Chromatography
Chemistry Coefficient of variation Physostigmine Extraction (chemistry) Biophysics Analytical chemistry Cell Biology Biochemistry High-performance liquid chromatography Fluorescence Working range Spectrometry Fluorescence Liquid–liquid extraction PHY Blood plasma Solvents Humans Molecular Biology Chromatography High Pressure Liquid |
| Zdroj: | Analytical biochemistry. 177(1) |
| ISSN: | 0003-2697 |
| Popis: | Physostigmine (PHY) is an anticholinergic drug used in the treatment of neuromuscular disorders and organophosphate poisoning. We described a sensitive, accurate, and reproducible method for PHY determination in biological materials. The method utilized a liquid/liquid, ion pair extraction, normal phase HPLC separation, and fluorometric quantitation at 240 nm excitation and 360 nm emission wavelength. We used neostigmine as a stabilizing agent to protect PHY from degradation and dimethylphysostigmine as an internal standard. The peak-height ratio vs concentration was linear over a working range from 0.50 to 25.0 ng/ml of PHY in plasma. Sensitivity of the method was 100 pg/ml of plasma which was the limit of quantitative detection under the experimental conditions used. Precision of the method was evaluated using plasma spiked with two concentrations of PHY: 1.0 and 10.0 ng/ml. Intra-day coefficient of variation (CV) ranged from 3.8 to 5.3%, and inter-day CV ranged from 1.8 to 3.6% for the two levels. The average recovery was 92%. We applied the method to examine the stability of PHY in plasma stored at -15 and -80 degrees C. The data indicated that PHY can be stored at either temperature for 9 weeks without undergoing significant alterations. |
| Databáze: | OpenAIRE |
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