A high-throughput HILIC-UHPLC-MS/MS approach for multi-tissue profiling of alkaloids in Papaver somniferum: Method development and chemotypic prediction

Abstract

A rapid and sensitive HILIC-UHPLC-MS/MS method was developed and validated for the simultaneous quantification of benzylisoquinoline alkaloids in Papaver somniferum tissues. The method fully separates six major alkaloids within 4.5 min using only 5 mg of dry plant material, with high recovery, minimal matrix effects, and excellent reproducibility across seeds, leaves, and capsules. Application to 15 food-grade poppy cultivars revealed significant chemotypic variation in seeds, as supported by profiling sugars, fatty acids, and phenolics. Time-course analysis in leaves and capsules uncovered distinct tissue-specific metabolic trends, with leaves serving as biosynthetic sources and capsules as primary sinks of alkaloids. Leaf metabolites at week 13 strongly predicted final alkaloid content in capsules, offering a non-destructive strategy for early screening. This high-throughput workflow supports chemotyping, cultivar screening, and quality control in food-grade poppy. Its sensitivity and low input make it ideal for both research and regulatory monitoring of alkaloids in food products.