Borate complexed sugars, sugar phosphates, and nucleotides present in tissue extracts were separated and quantitated in 4 hr. An anion-exchange resin column and a programmed borate/acetate buffer gradient were used. Sugar residues were determined by a very sensitive orcinol/H,SO, reaction. Samples r
Determination of inositol phosphates and other biologically important anions by ion chromatography
โ Scribed by Robert E. Smith; Ronald A. MacQuarrie
- Publisher
- Elsevier Science
- Year
- 1988
- Tongue
- English
- Weight
- 583 KB
- Volume
- 170
- Category
- Article
- ISSN
- 0003-2697
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โฆ Synopsis
Ion chromatography has been applied to the simultaneous, multi-component determination of biologically important anions. More than 20 different biologically important anions were separated on high performance ion-exchange columns and detected using chemically suppressed conductivity. Application of the technique to the separation of inositol mono-, bis-, and trisphosphates shows that these compounds can be separated from the other ions tested and can be detected at concentrations that may be found in vivo. For inositol monophosphate, the conductivity was proportional to the amount of compound from less than 20 pmol to more than 400 nmol. Although alternative methods are available for assaying each of these anions individually, the advantages of ion chromatography lie in the sensitivity of detection, the speed of separation, and the ability to simultaneously determine numerous ions. This method should be broadly applicable to studies of second messengers, measurements of reaction rates, and various metabolic studies.
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We have developed an ion-pair reverse-phase HPLC method to measure inositol phosphates in 32P-labeled cells. The different chromatographic parameters were analyzed to optimize the resolution of the 32P-labeled metabolites. Analysis of inositol phosphates in biological samples was improved by a singl
Of the ion-chromatographically distinct isomers, 11 of the 12 myo-inositol trisphosphates (IP,) and all of the 9 myo-inositol bisphosphates (IP,) have been found in a chemical hydrolysate of phytic acid. The isomers were fractionated by anion-exchange chromatography by a method that is suitable for