Highly sensitive HPLC method for the determination of galantamine in human plasma and urine through derivatization with dansyl chloride using fluorescence detector

• Published in: Luminescence (Chichester, England) England), 2017-11, Vol.32 (7), p.1145-1149
• Main Authors: Özdemir, Elif, Tatar Ulu, Sevgi
• Format: Article
• Language: English
• Subjects: Acetonitrile; Blood plasma; Chloride; Chlorides; Chromatography; Chromatography, High Pressure Liquid - methods; dansyl chloride; Dansyl Compounds - chemistry; derivatization; Detection; Dichloromethane; Fluorescence; Galantamine; Galantamine - blood; Galantamine - urine; High performance liquid chromatography; HPLC; Humans; Ions; Limit of Detection; Linearity; Liquid chromatography; Mathematical analysis; Methods; Naphthalene; pH effects; Phosphoric acid; Plasma; Plasmas (physics); Reproducibility; Reproducibility of Results; Retention time; Sensitivity and Specificity; Standard deviation; Urine; validation
• ISSN: 1522-7235; 1522-7243
• DOI: 10.1002/bio.3301

A new method based on fluorescence derivatization with 5‐(dimethylamino) naphthalene‐1‐sulfonyl chloride (dansyl chloride) was developed for the quantitative determination of galantamine in human plasma and urine using high‐performance liquid chromatography. The reaction between galantamine and dansyl chloride was optimally realized in 30 min at room temperature and pH 10.5, with a reagent to galantamine molar ratio of 2.13. The derivative was extracted with dichloromethane, and the extract was dried under a nitrogen stream and dissolved in the mobile phase. Chromatographic analysis was performed with an Inertsil C18 column and a mobile phase comprising 40% acetonitrile and 60% 10 mM o‐phosphoric acid, 1.2 ml/min. The injection volume was 20 μl. The derivatives were detected with a fluorescence detector (excitation 375 nm/emission 537 nm). The retention time for the dansyl derivative of galantamine was 16.8 min. Linearity was observed between 125 and 2000 ng/ml in water, urine and plasma. The limit of detection and limit of quantification for the developed method were 6.27–70.99 and 18.81–212.97 ng/ml, respectively. Per cent recovery was calculated as 95.15 for urine and 95.78 for plasma. Interday repeatability values for urine and plasma samples (n = 6) at three different concentrations were calculated as a per cent relative standard deviation of 0.24–0.59 and 0.35–0.56. The corresponding per cent relative standard deviation values for intraday repeatability were 0.13–0.51 and 0.04–0.15, respectively.