Determination of mitomycin C, 2,7-diaminomitosene, 1,2-cis- and 1,2-trans-1-hydroxy-2,7-diaminomitosene in tumour tissue by high-performance liquid chromatography.
A high-performance liquid chromatographic method is described for the determination of mitomycin C (MMC) and its metabolites 2,7-diaminomitosene (2,7-DM), 1,2-cis-1-hydroxy-2,7-diaminomitosene (cis-hydro) and 1,2-trans-1-hydroxy-2,7-diaminomitosene (trans-hydro) in tumour tissue. N-la-Methylmitomycin C (porfiromycin, PM) was used as an internal standard. Two factors were critical in resolving the metabolites: pH and buffer ionic strength, where the retention times of the four components were affected in the order 2,7-DM >> cis-hydro >> trans-hydro >> MMC. The optimal isocratic conditions (flow-rate 1 ml/min) were 18 mM sodium phosphate pH 5.8-methanol (74:26) and a column temperature of 40 degrees C on a Spherisorb ODS-2 column (25 cm x 4.6 mm I.D.). Liquid-liquid extraction [twice with chloroform-propan-2-ol-ethyl acetate (2:2:1)] is described for tumour tissue. Recoveries varied depending on the component: MMC, 71.9 +/- 12.4%; PM, 85.5 +/- 27%; 2,7-DM, 51.7 +/- 5.4%; cis-hydro, 52.0 +/- 16.8%; trans-hydro, 62 +/- 8%. When applied to the analysis of a rat mammary carcinoma treated intra-tumourally with 450 micrograms of MMC five drug-related "metabolite" peaks were detected. Three of these co-chromatographed with standards of 2,7-DM, cis- and trans-hydro, and had identical absorption maxima to their respective standards, with the possible exception of trans-hydro.
Cummings
J
J
Imperial Cancer Research Fund, Medical Oncology Unit, Western General Hospital, Edinburgh, UK.
Chirrey
L
L
Willmott
N
N
Halbert
G W
GW
Smyth
J F
JF
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006863:Hydrogen-Ion Concentration; D008325:Mammary Neoplasms, Experimental / Q000378:metabolism*; D016685:Mitomycin / Q000032:analysis*; D008937:Mitomycins / Q000032:analysis*; D051381:Rats; D013056:Spectrophotometry, Ultraviolet
D008937:Mitomycins; D016685:Mitomycin; C056707:2,7-diaminomitosene; C049686:1-hydroxy-2,7-diaminomitosene