TY - JOUR
T1 - A comparison of the efficiency of nucleotide extraction by several procedures and the analysis of nucleotides from extracts of liver and isolated hepatocytes by HPLC
AU - Rias, T. L.
AU - Zorich, N. L.
AU - Williams, M. D.
AU - Richardson, A.
N1 - Funding Information:
ACKNOWLEDGMENTS We would like to thank Dr. Daniel Paschal for his helpful assistance fa developing the ?PLC system used fa this study. This work was supportod in part by NIH grant 1 ROl AG 00344-02.
PY - 1980/1/1
Y1 - 1980/1/1
N2 - A high pressure liquid chromatography (HPLC) system was devel-oped, capable of resolving most 5’-nucleotides and nucleotide-sugars present in liver tissue. Using three different extraction procedures, the recovery of the twelve major 5’-ribonucleotides from solutions of nucleotide standards, or liver samples, or isolated hepatocytes was compared. Nucleotides were obtained from acid extracts for HPLC analysis by adsorbing the nucleotides on charcoal, or precipitating the acid (perchloric acid) by KOH, or extracting the acid with alamine/ freon. The last two procedures were found to be superior to the charcoal adsorption procedure for recovering nucleotides from acid extracts. The recovery of nucleotides from either liver samples or isolated hepatocytes by these two procedures was similar; however, the recovery of nucleotides from standard solutions was slightly higher for the alamine/freon procedure than the KOH precipitation procedure.
AB - A high pressure liquid chromatography (HPLC) system was devel-oped, capable of resolving most 5’-nucleotides and nucleotide-sugars present in liver tissue. Using three different extraction procedures, the recovery of the twelve major 5’-ribonucleotides from solutions of nucleotide standards, or liver samples, or isolated hepatocytes was compared. Nucleotides were obtained from acid extracts for HPLC analysis by adsorbing the nucleotides on charcoal, or precipitating the acid (perchloric acid) by KOH, or extracting the acid with alamine/ freon. The last two procedures were found to be superior to the charcoal adsorption procedure for recovering nucleotides from acid extracts. The recovery of nucleotides from either liver samples or isolated hepatocytes by these two procedures was similar; however, the recovery of nucleotides from standard solutions was slightly higher for the alamine/freon procedure than the KOH precipitation procedure.
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U2 - 10.1080/01483918008060160
DO - 10.1080/01483918008060160
M3 - Article
AN - SCOPUS:0001215381
VL - 3
SP - 133
EP - 158
JO - Journal of Liquid Chromatography and Related Technologies
JF - Journal of Liquid Chromatography and Related Technologies
SN - 1082-6076
IS - 1
ER -