For the determination of vitamin B 6 derivatives, a high performance liquid chromatography(HPLC) system composed of a single reversed-phase octadecylsilane column and fluorescence detector with an isocratic elution wa...For the determination of vitamin B 6 derivatives, a high performance liquid chromatography(HPLC) system composed of a single reversed-phase octadecylsilane column and fluorescence detector with an isocratic elution was proposed. This HPLC system perfectly analyzed pyridoxine, pyridoxal, pyridoxamine, pyridoxine 5′-phosphate and pyridoxamine 5′-phosphate together with pyridoxic acid in biological samples. In this method, relatively low fluorescence intensity of pyridoxal 5′-phasphate was improved by converting it to pyridoxic 5′-phosphate with KCN. The analytical procedure is very simple and economical, it is a useful tool for studying vitamin B 6 in animal body.展开更多
Stroma--free hemoglobin (SFHb) was prepared from fresh or out--dated whole blood by lysing red cells. At different temperatures, pyridoxal 5--phosphate (PLP) was added in a 4:1 molar-ratio to the Hb tetramer followed ...Stroma--free hemoglobin (SFHb) was prepared from fresh or out--dated whole blood by lysing red cells. At different temperatures, pyridoxal 5--phosphate (PLP) was added in a 4:1 molar-ratio to the Hb tetramer followed by reduction with NaBH4 under N2 for 16 hrs subsequenl crosslinking by glutaraldehyde in presence of lysine for 14--16 hrs. Yielded soluble pyridoxalated polyhemoglobin (Poly Hb--P). 5%--20% gradient SDS--PAGE showed the change of the Hb molecular weight (MW) after crosslinking. Gel chromatography showed that the poly Hb--P had a continuous distribution of molecular weight with the range from about 65,000 to 600,000 Dalton. The poly Hb--P solution provided a normal oxygen carrying with 23.1 of P50, at pH 7.24 or 26.1 of P50 at pH 7.24展开更多
文摘For the determination of vitamin B 6 derivatives, a high performance liquid chromatography(HPLC) system composed of a single reversed-phase octadecylsilane column and fluorescence detector with an isocratic elution was proposed. This HPLC system perfectly analyzed pyridoxine, pyridoxal, pyridoxamine, pyridoxine 5′-phosphate and pyridoxamine 5′-phosphate together with pyridoxic acid in biological samples. In this method, relatively low fluorescence intensity of pyridoxal 5′-phasphate was improved by converting it to pyridoxic 5′-phosphate with KCN. The analytical procedure is very simple and economical, it is a useful tool for studying vitamin B 6 in animal body.
文摘Stroma--free hemoglobin (SFHb) was prepared from fresh or out--dated whole blood by lysing red cells. At different temperatures, pyridoxal 5--phosphate (PLP) was added in a 4:1 molar-ratio to the Hb tetramer followed by reduction with NaBH4 under N2 for 16 hrs subsequenl crosslinking by glutaraldehyde in presence of lysine for 14--16 hrs. Yielded soluble pyridoxalated polyhemoglobin (Poly Hb--P). 5%--20% gradient SDS--PAGE showed the change of the Hb molecular weight (MW) after crosslinking. Gel chromatography showed that the poly Hb--P had a continuous distribution of molecular weight with the range from about 65,000 to 600,000 Dalton. The poly Hb--P solution provided a normal oxygen carrying with 23.1 of P50, at pH 7.24 or 26.1 of P50 at pH 7.24
