تفاعل #336576

معادلة التفاعل

L-arginine

L-lysine

D-ribose

→

Pentosidine

ظروف التفاعل

المعالجة

1. 1
   workup.ADDITIONThe cooled mixture was poured onto a Buchner funnel
2. 2
   workup.ADDITIONfilled with Dowex 50×4-400 ion-exchange resin (Aldrich Chem. Co., Inc., Milwaukee, Wisc.)
3. 3
   غسيلThe resin was washed with 2 liters each of water and 1 M pyridine
4. 4
   غسيلfollowed by elution of pentosidine-
5. 5
   workup.ADDITIONcontaining material with 1 liter of 2 N NaOH
6. 6
   تركيزconcentrated by rotary evaporation
7. 7
   أخرىpurified by HPLC
8. 8
   أخرىThe material was also chromatographed on Whatman 17 Chr paper (Whatman Inc., Clifton, N.J.)
9. 9
   غسيلUpon elution from the paper
10. 10
    أخرىcollected by reversed-phase

الإجراء التجريبي

Three liters containing 100 mM each of L-arginine, L-lysine and D-ribose, at pH 7.3, Were heated for 1 hr at 80° C. The cooled mixture was poured onto a Buchner funnel filled with Dowex 50×4-400 ion-exchange resin (Aldrich Chem. Co., Inc., Milwaukee, Wisc.) equilibrated according to conditions of Boas (Boas, N. F., J. Biol. Chem., 204, pp. 553-563, 1953). The resin was washed with 2 liters each of water and 1 M pyridine, followed by elution of pentosidine-containing material with 1 liter of 2 N NaOH. The material was then adjusted to pH 7.4 with HCl, concentrated by rotary evaporation, passed through a Bio-Gel P-2 column and purified by HPLC as described above. The material was also chromatographed on Whatman 17 Chr paper (Whatman Inc., Clifton, N.J.) using 1:1 water/pyridine. Upon elution from the paper, the material was reinjected and collected by reversed-phase HPLC using TFA as the counterion.

المصدر