Fluorometric high-performance liquid chromatographic analysis of 10-deazaaminopterin, 10-ethyl-10-deazaaminopterin, and known metabolites Journal Article
| Authors: | Kinahan, J. J.; Samuels, L. L.; Farag, F.; Fanucchi, M. P.; Vidal, P. M.; Sirotnak, F. M.; Young, C. W. |
| Article Title: | Fluorometric high-performance liquid chromatographic analysis of 10-deazaaminopterin, 10-ethyl-10-deazaaminopterin, and known metabolites |
| Abstract: | The antifolate compounds 10-deazaaminopterin (10-dAM) and 10-ethyl-10-deazaaminopterin (10-EdAM) are therapeutically superior to methotrexate in transplanted murine tumor systems and in human tumor xenografts growing in immunodeficient "nude" mice. The increased therapeutic index of these analogs correlates with their selective uptake, retention, and polyglutamation within neoplastic cells. We have developed a fluorescence high-performance liquid chromatographic assay applicable to 10-dAM, 10-EdAM, their polyglutamate anabolites, and their 7-hydroxy (7-OH) and deglutamate catabolites. The assay is based upon the high native fluorescence of pteridine-containing compounds which contain carbon in the 10 position. The assay employs a reverse-phase C-18 column and an ascending acetonitrile gradient in 50 mm phosphate, pH 7.0. The compounds are extracted from plasma and urine with 95 ± 7% and 98 ± 2% recoveries, respectively, using C-18 Sep-Paks. The linear range of the assay is, for 10-dAM, 2-10 nm, and for 10-EdAM, 1-100 nm. Polyglutamated metabolites of [3H]10-EdAM isolated from L1210 cells have been separated by HPLC with identification of five derivatives (Glu 1-5) confirmed by enzymatic peak shift using serum conjugase and by quantitative correlation of fluorescence intensity, radioactivity, and titration inhibition of dihydrofolate reductase. The assay has been used successfully in pharmacokinetic analyses of plasma and urine samples from patients receiving 10-dAM and 10-EdAM. In patients who had received 10-EdAM, 7-OH-10-EdAM, and the deglutamate catabolite were also detected. This HPLC fluorescence assay is superior to the dihydrofolate reductase inhibition and binding assays with regard to specificity and precision; moreover, it can provide a means for simultaneous assay of the physiologically important anabolites and catabolites of these new antifolates. © 1985. |
| Keywords: | cancer chemotherapy; controlled study; unclassified drug; human cell; drug efficacy; methotrexate; methodology; neoplasms; animals; mice; spectrometry, fluorescence; mus musculus; mice, nude; murinae; drug monitoring; drug metabolism; drug absorption; drug blood level; high performance liquid chromatography; chromatography, high pressure liquid; radioisotope; drug determination; therapy; aminopterin; urinary tract; intravenous drug administration; drug urine level; human experiment; fluorometry; antifolate; 10 deazaaminopterin; drug comparison; polyglutamic acid; leukemia l1210; edatrexate; humans; human; priority journal; article; 10 ethyl 10 deazaaminopterin deglutamate; blood and hemopoietic system; methotrexate h 3; drug analysis; 10 ethyl 7 hydroxy 10 deazaaminopterin; fluorescence hplc; 10 deazaaminopterin polyglutamate; 10 ethyl 10 deazaaminopterin h 3 |
| Journal Title: | Analytical Biochemistry |
| Volume: | 150 |
| Issue: | 1 |
| ISSN: | 0003-2697 |
| Publisher: | Academic Press, Elsevier Inc |
| Date Published: | 1985-10-01 |
| Start Page: | 203 |
| End Page: | 213 |
| Language: | English |
| DOI: | 10.1016/0003-2697(85)90460-9 |
| PROVIDER: | scopus |
| PUBMED: | 2417507 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 26 October 2021 -- Source: Scopus |
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