Preparative-scale separation of colistin sulphate bulk sample was carried o
ut on a preparative poly(styrene-divinylbenzene) stationary phase. Isocrati
c elution with acetonitrile-sodium sulphate solution (0.7%.m/v; pH adjusted
to 2.5 with TFA) - water (16 : 50 : 34, % v/v/v) was carried out at a flow
rate of 4.0ml min(-1). Six colistin components were isolated and character
ized using H-1 and C-13 NMR. The molecular weights were confirmed by mass s
pectrometry. The structures of 2 components were determined for the first t
ime. Polymyxin E-7 was identified as having the same composition as polymyx
in E-1, except that the fatty acid moiety was 7-methyloctanoic acid. Isoleu
cine polymyxin E-8 was characterized as having the same composition as isol
eucine polymyxin E-1 with 7-methylnonanoic acid as the fatty acid moiety.