This method uses precolumn and reverse phase C18 column and UV-VIS detector with deuterium lamp, with a mobile phase consisting of 80% acetonitrile, 3% methanol (100 mM ammonium acetate, 0.1% triethylamine), and 15% dioxan
. The mobile phase is pumped through the system by a flow of 1.5 mL/min with maximum pressure of 350 Kgf and the minimum of 0.
After dehydration in a graded series of ethanol, the specimens were infiltrated with dioxan
To 5 mg of F-21-HS, 200 [micro]L of dimethyl formamide, 200 [micro]L of dioxan
, and 100 [micro]L of distilled water containing 10 mg of N-hydroxysuccinimide and 20 mg of 1-ethyl-3-(3-dimethyl-amino-propyl) carbodiimide-HCL were added; the reaction mixture was vortex-mixed and kept overnight at 4 [degrees]C.
Ar solvent ([degrees]C) 3 -- EtOH 226-229 4a 4-OH[C.sub.6][H.sub.4] DMF/EtOH 248-250 4b 4-MeO[C.sub.6][H.sub.4] DMF/EtOH 255-257 4c 4-OH,3-MeO[C.sub.6][H.sub.3] DMF/EtOH 240-243 5a 4-OH[C.sub.6][H.sub.4] DMF 271-273 5b 4-MeO[C.sub.6][H.sub.4] DMF 282-284 5c 4-OH,3-MeO[C.sub.6][H.sub.3] DMF 266-268 6 -- Dioxan
236-239 7 -- EtOH 244-246 8a 4-OH[C.sub.6][H.sub.4] DMF/EtOH 273-275 8b 4-Cl[C.sub.6][H.sub.4] DMF/EtOH 264-267 8c 4-[H.sub.2]NS[O.sub.2] DMF/EtOH 283-285 [C.sub.6][H.sub.4] Comp.
To a solution of 2-thioxopyrimidin (4e) (1 mmol) and ethyl 2-bromoacetate (1 mmol) was refluxed for 6 h in dioxan
(10 mL) /presence of catalytic amount of pyridine (1mL).