High-performance thin-layer chromatography combined with effect-directed assays and high-resolution mass spectrometry as an emerging hyphenated technology: A tutorial review

Among the analytical advances, hyphenated HPTLC offers great potential for solving pressing questions. It provides straightforward information about effects arising from individual compounds in complex or natural samples separated in parallel. The chromatographic separation is combined with effect-directed detection using enzymatic or biological assays. This helps to select from the thousands of compounds in a sample the important ones that need to be further characterized using high-resolution mass spectrometry. Unique benefits are discussed exemplarily arising from its super-hyphenation, minimum requirements for sample preparation, detection of multi-modulating compounds or agonistic versus antagonistic effects, and miniaturized on-surface metabolization. HPTLC stands for a versatile, creative and flexible open-format technique. As miniaturized open-source LabToGo system, it shows the potential to be applied as Citizen Science.