[Combinations of liquid chromatography methods for the breeding of high alkaloid-containing poppies]

Abstract

An analytical strategy for the determination of alkaloid content of poppy capsules was developed for the efficient plant breeding. The first step of the strategy is a screening by circular multilayer (ML) overpressured layer chromatography (OPLC) with large number (28 samples/chromatoplate) of samples under UV (254 and 366 nm) light. The next step is a TLC separation with densitometric evaluation for quantitative determination of the selected samples (ca. 50% of all samples). These results are confirmed by ionpair reversed-phase (RP) high performance liquid chromatography (HPLC) for the most interesting (20% of all samples) poppy capsules. If the difference of the two applied quantitative methods is higher than 10% the investigations have to be repeated by new mobile phase systems with different solvent strength and selectivity factor values to eliminate the disturbing effect of the minor compounds which appear during the plant breeding. On the basis of the analytical results two new populations were formed, one of them ("MAMOR") has high morphine (approximately 20 mg/g) content, the other one ("TEMAX") is characterized by ability of tebain (approximately 16 mg/g) accumulation. The two new poppy sorts are verification for the effectiveness of the combination of different separation techniques (OPLC, TLC, HPLC) and UV detection modes (off-line and on-line) for medicinal plant breeding.