High-performance liquid chromatographic assay validation of Manumycin A in mouse plasma.

Abstract

Manumycin A is a natural antibiotic produced by Streptomyces parvulus that has antineoplastic activity against a variety of human cancers in nude mouse models. We have developed a highly sensitive reverse phase high-performance liquid chromatography (HPLC) method based on ultraviolet (UV) detection for the determination of manumycin A in mouse plasma. Manumycin A was isolated from mouse plasma by solid-phase extraction. A gradient elution of methanol and 0.05 M H(3)PO(4) with 0.2% triethylamine mobile phase was employed and separation was achieved with a C(18) analytical column. Manumycin A was detected by UV absorption at 345 nm. Retention time for manumycin A was 8.9+/-0.2 min. The manumycin A peak was baseline resolved, with the nearest peak at 1.5 min distance and no interfering peaks detected. Inter- and intra-day coefficients of variance were less than 6.1 and 5.1%, respectively. Based on an extracted manumycin A standard plasma sample of 0.25 microg/ml, the assay precision was 99.8% with a mean accuracy of 95.1%. At plasma concentrations of 0.5 and 5 microg/ml, the mean recovery rates of manumycin A were 59.64 and 60.28%, respectively. The lower limit of detection (LLD) for manumycin A was 0.1 microg/ml in mouse plasma. The lower limit of quantification (LLQ) for manumycin A was 0.125 microg/ml. Results of the stability study indicated that when frozen at -80 degrees C, manumycin A was stable in mouse plasma for up to 2 weeks. This method is useful in quantification of manumycin A in mouse plasma for clinical pharmacology studies in mice.

• Authors: Joanne Gonzales, Sai-Ching Jim Yeung, Judith A Smith
• Journal: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (J Chromatogr B Analyt Technol Biomed Life Sci) Vol. 776 Issue 2 Pg. 177-82 (Sep 05 2002) ISSN: 1570-0232 [Print] Netherlands
• PMID: 12137999 (Publication Type: Journal Article, Validation Study)
• Copyright: Copyright 2002 Elsevier Science B.V.

Chemical References

• Anti-Bacterial Agents
• Polyenes
• Polyunsaturated Alkamides
• manumycin

Topics

• Animals
• Anti-Bacterial Agents (blood)
• Chromatography, High Pressure Liquid (methods)
• Mice
• Polyenes (blood)
• Polyunsaturated Alkamides
• Spectrophotometry, Ultraviolet