Journal article
High-pressure liquid chromatography
Analytical biochemistry, Vol.90(1), pp.289-299
1978
DOI: 10.1016/0003-2697(78)90033-7
Abstract
A high-pressure liquid column chromatographic method has been developed for the separation and identification of ribonucleosides, deoxyribonucleosides, and bases. This method is capable of detecting these components at 1.0 to 5.0 ng applied; is reproducible under conditions of constant pressure, temperature, and pH; and is rapid, requiring about 30 min for a complete chromatographic separation of ribonucleosides from deoxyribonucleosides and from bases. These separations were carried out under different pH values and buffers, namely, phosphate buffer containing 2.5% methanol at pH 6.9 and 3.0 or 50 m
m sodium borate buffer, pH 9.0. These different conditions were utilized to obtain more definitive identification and quantitation of normal metabolites and their counterparts, the antimetabolites. The advantage of this method is that the 20 naturally occurring components are separated from each other by an isocratic elution method, alleviating the need for a gradient elution system, which produces a drift in the baseline with increasing concentration of the eluting buffer, especially when the instrument is operating at maximum sensitivity, thus hindering the quantitation of the separated components. The potential application of this method for the quantitation of plasma metabolites and antimetabolites such as Ara-C
2, Ara-U, and F-pyrimidine is describe.
Details
- Title: Subtitle
- High-pressure liquid chromatography
- Creators
- Youcef M. Rustum - Roswell Park Cancer Institute
- Resource Type
- Journal article
- Publication Details
- Analytical biochemistry, Vol.90(1), pp.289-299
- Publisher
- Elsevier Inc
- DOI
- 10.1016/0003-2697(78)90033-7
- ISSN
- 0003-2697
- eISSN
- 1096-0309
- Language
- English
- Date published
- 1978
- Academic Unit
- Internal Medicine; Hematology, Oncology, and Blood & Marrow Transplantation
- Record Identifier
- 9984359860402771
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