Highly sensitive HPLC method for the determination of galantamine in human plasma and urine through derivatization with dansyl chloride using fluorescence detector
Luminescence, cilt.32, sa.7, ss.1145-1149, 2017 (SCI-Expanded, Scopus)
- Yayın Türü: Makale / Tam Makale
- Cilt numarası: 32 Sayı: 7
- Basım Tarihi: 2017
- Doi Numarası: 10.1002/bio.3301
- Dergi Adı: Luminescence
- Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
- Sayfa Sayıları: ss.1145-1149
- Anahtar Kelimeler: dansyl chloride, derivatization, HPLC, plasma, urine, validation
- İstanbul Yeni Yüzyıl Üniversitesi Adresli: Evet
Özet
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.