Rapid and selective isolation of radiolabelled inositol phosphates from cancer cells using solid-phase extraction.
A method is described for rapid and selective determination of radiolabelled inositol phosphates in cancer cells using solid-phase extraction with Bond Elut strong anion-exchange minicolumns. The inositol phosphates IP1, IP2 and IP3 are selectively eluted with 0.05, 0.3 and 0.8 M ammonium formate-0.1 M formic acid, respectively. Cancer cells are extracted with 10% perchloric acid which is then neutralised prior to loading samples on to the minicolumns. Recovery is 54.1, 66.6 and 61.3% for IP1, IP2 and IP3 with between-day coefficients of variation of 7.6, 6.8 and 1.9%, respectively. When the method was applied to cancer cells high-performance liquid chromatographic analyses confirmed both the identity of the IP1, IP2 and IP3 fractions and showed that there was no detectable cross contamination of these inositol phosphates with each other.
Anderson
L
L
Imperial Cancer Research Fund, Medical Oncology Unit, Western General Hospital, Edinburgh, UK.
Cummings
J
J
Smyth
J F
JF
D002851:Chromatography, High Pressure Liquid; D007295:Inositol Phosphates / Q000302:isolation & purification*; D010911:Pituitary Neoplasms / Q000378:metabolism*; D015203:Reproducibility of Results; D014316:Tritium; D014407:Tumor Cells, Cultured
D007295:Inositol Phosphates; D014316:Tritium