Simultaneous liquid chromatographic assay of amantadine and its four related compounds in phosphate-buffered saline using 4-fluoro-7-nitro-2,1,3-benzoxadiazole as a fluorescent derivatization reagent

Abstract

Simultaneous HPLC assay of 1-adamantanamine hydrochloride (amantadine) and its four related compounds [2-adamantanamine hydrochloride (2-ADA), 1-adamantanmethylamine (ADAMA), 1-(1-adamantyl)ethylamine hydrochloride (rimantadine) and 3,5-dimethyl-1-adamantanamine hydrochloride (memantine)] in phosphate-buffered saline (pH 7.4) after pre-column derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) was developed. Phosphate-buffered saline samples were mixed with borate buffer and NBD-F solution in acetonitrile at 60 degrees C for 5 min and injected into HPLC. Five derivatives were well separated from each other. The lower limits of detection of amantadine, 2-ADA, ADAMA, rimantadine and memantine were 0.008, 0.001, 0.0008, 0.0015 and 0.01 microg/mL, respectively. The coefficients of variation for intra- and inter-day assay were less than 6.4 and 8.2%, respectively. The method presented was applied to a binding study of these compounds to human alpha(1)-acid glycoprotein. While affinity constants and capacities for ADAMA, rimantadine and memantine were calculated by means of Scatchard plots, those for the others were not determined. ADAMA, rimantadine and memantine were bound with different affinities and capacities. These results indicate that NBD-F is a good candidate as a fluorescent reagent to simultaneously determine amantadine and its four related compounds by HPLC after pre-column derivatization. Our method can be applied to binding studies for protein.

High‐performance liquid chromatographic determination of memantine hydrochloride in rat plasma using sensitive fluorometric derivatization

“…Most HPLC methods involved dansyl chloride [4], o-phthalaldehyde (OPA) [10,11], 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) [12] and 1-fluoro-2,4-dinitrobenzene (DNFB) [21] as derivatization reagents to react with tricyclic amines. In this study, dansyl chloride, 4-fluoro-7-nitro-2,1,3-benzoxadiazole and 1-fluoro-2,4-dinitrobenzene were tested to react with memantine hydrochloride and we found that FMOC-Cl derivates of memantine hydrochloride had advantages of higher sensitivity, less interference, and simpler reaction conditions than others.…”

“…1) for UV or fluorimetric detection. The assay of tricyclic amines is always performed using precolumn [4,[10][11][12] derivatization methods, or other detections, MS [13,14]. And memantine hydrochloride also has been determined by GC-MS [15] and UPLC [16].…”

Determination of Amantadine Residue in Honey by Solid‐phase Extraction and High‐performance Liquid Chromatography with Pre‐column Derivatization and Fluorometric Detection

“…Solid-phase extraction (SPE) is a simple, efficient, and solvent-reducing alternative that has been successfully used to analyze various chemical residues in honey samples. [21][22][23] In light of the procedures for derivatization of AMA with NBD-F of Higashi et al(2006), 16 we made a slight modification by dissolving the sample in borate buffed and with fluorescence detection at an excitation wavelength of 470 nm and an emission wavelength of 530 nm. When these conditions were applied in our research, the longest retention times of the AMA and IS derivatives were obtained with 16.9 min and 30.2 min, respectively.…”

“…16 Briefly, each 400 µL solution for the calibration or extracted sample solution was transferred to an HPLC brown glass vial, and then 100 µL NBD-F solution (21.8 mmol/L in acetonitrile, stored at -20 ℃, restored to room temperature before use) was added in. The mixture was allowed to react for 12 min at 60 ℃ and then placed on ice to stop the derivatization reaction.…”

“…Several derivatizing reagents have been intensively studied for HPLC analysis of AMA, including o-phthalaldehyde and 1-thio-β-D-glucose, 11 (2-naphthoxy) acetyl chloride, 12 anthraquinone-2-sulfonyl chloride, 13 dansyl chloride, 14 1-fluoro-2,4-dinitrobenzene, 15 and 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F). 16 Among those, NBD-F selectively reacts with the primary and secondary amino groups and has been successfully used for the analyses of many compounds. 14,[17][18][19] In this paper an analytical method was developed to analyze AMA residue in honey.…”