High-throughput analytical strategy with combined planar and column liquid chromatography for improvement of the poppy (Papaver somniferum L.) with a high alkaloid content

Abstract

Poppy capsules with a high alkaloid content are of great importance to the pharmaceutical industry, because approximately 35 000 tons of straw (capsule with short stem), 350 tons of straw concentrate, and 1000 tons of opium are used annually for extraction of morphinane alkaloids. The combined use of four different liquid chromatographic methods is proposed for determination of alkaloid content. In the first, semi-quantitative, method screening is performed by multilayer overpressured-layer chromatography (MLOPLC) in which 142 samples are separated on silica as stationary phase within 5 s per sample. If the morphine content is >1.3% it is then measured quantitatively by NPHPTLC (normal-phase high-performance thin-layer chromatography) combined with densitometric evaluation. A second, quantitative, determination is always performed by means of a rapid RPHPLC (reversed-phase high-performance liquid chromatography) method in which the separation time is less than 4 min per sample. If the difference between the alkaloid content measured by use of the last two methods is >12% a confirmatory RPHPLC method with increased selectivity and analysis time (15 min) must be used. The high throughput strategy presented has been used for analysis of ca. 15 000 samples per year, per genotype. Systematic selection, cross-breeding, and this analytical strategy with combined planar and column liquid chromatographic methods resulted in the discovery of two new candidate cultivars with high morphine (ca. 2%) and thebaine (ca. 1.5%) content.