(1) RAPID HIGH PERFORMANCE LIQUID CHROMATOGRAPHY OF CYCLOSPORIN A, CIMETIDINE, CYCLOPHOSPHAMIDE, 5-FLUOROURACIL AND 5-AZACYTIDINE. (2) INJECTION OF SOLUTES IN STRONGER SOLVENTS THAN THE MOBILE PHASE IN LIQUID CHROMATOGRAPHY

ABU MOHAMMAD RUSTUM, Marquette University

Abstract

This research has developed simple and rapid analytical methods to quantitate the drugs 5-Azacitidine, Cimetidine, Cyclophosphamide, Cyclosporin A and 5-Fluorouracil from biological samples by using HPLC. 5-Azacitidine is isolated from plasma with acetonitrile and zinc sulfate. The solution is vortexed, centrifuged, filtered, and chromatographed with a PRP-1 column with 74:26, 10mM sodium octanesulfonate:MeOH mobile phase. The drug is detected by UV absorption at 266nm. The time of assay is 15 minutes. The detection limit is 30ng/mL with %RSD of 6. Cimetidine is extracted after precipitation of the proteins with MeOH. After vortexing, centrifuging, and salting out, the MeOH solution is chromatographed at 210nm with a PRP-1 column with mobile phase of 87.5:12.5, 20mM K(,2)HPO(,4):MeOH. The assay time is less than 20 minutes. Cyclophosphamide is isolated after precipitation of proteins with acetonitrile. The solution is vortex, centrifuged, salted out with K(,2)CO(,3), and chromatographed with an ODS column with mobile phase of 80:20, 10mM phosphate buffer:acetonitrile. The drug is detected at 190nm. The time of assay is less than 25 minutes. Cyclosporin A is extracted with acetonitrile. The solution is vortexed, centrifuged, and the acetonitrile is salted out with (NH(,4))(,2)SO(,4). This solution is treated with cation and anion exchange resins, washed in hexane, and chromatographed with an ODS column with mobile phase of 70:30, acetonitrile:water. Cyclosporin A is monitored at 200nm. The assay time is 30 minutes. After precipitation of the proteins 5-fluorouracil is extracted with acetonitrile. The solution is vortexed, centrifuged, salted out with (NH(,4))(,2)SO(,4). The resulting solution is evaporated, reconstituted with an aliquot of the mobile phase, and chromatographed with a PRP-1 column with a 74:26, 10mM tetrabutylammonium hydroxide:MeOH mobile phase. The drug is monitored at 266nm. The assay time is about 30 minutes. In addition, experiments were done to determine the reason(s) for the deterioration of solute peak height with large injection volumes when the solvent strength of the sample and mobile phase differ greatly. The compartment theory developed in this research satisfactorily explains the above affect, and several solutions to the problem were found.

Recommended Citation

RUSTUM, ABU MOHAMMAD, "(1) RAPID HIGH PERFORMANCE LIQUID CHROMATOGRAPHY OF CYCLOSPORIN A, CIMETIDINE, CYCLOPHOSPHAMIDE, 5-FLUOROURACIL AND 5-AZACYTIDINE. (2) INJECTION OF SOLUTES IN STRONGER SOLVENTS THAN THE MOBILE PHASE IN LIQUID CHROMATOGRAPHY" (1986). Dissertations (1962 - 2010) Access via Proquest Digital Dissertations. AAI8618718.
http://epublications.marquette.edu/dissertations/AAI8618718

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