Abstract
Biochemical detection (BCD) methods are commonly used to screen plant extracts for specific biological activities in batch assays. Traditionally, bioactives in the most active extracts were identified through time-consuming bio-assay guided fractionation until single active compounds could be isolated. Not only are isolation procedures often tedious, but they could also lead to artifact formation. On-line coupling of BCD assays to high performance liquid chromatography (HPLC) is gaining ground as a high resolution screening technique to overcome problems associated with pre-isolation by measuring the effects of compounds post-column directly after separation. To date, several on-line HPLC-BCD assays, applied to whole plant extracts and mixtures, have been published. In this review the focus will fall on enzyme-based, receptor-based and antioxidant assays.
Highlights
Plants have provided the basic building blocks for a number of highly effective drugs and they remain an attractive option for discovery of new molecular entities, due to their still largely untapped chemical diversity [1]
Plasma samples were analyzed for ligands of urokinase plasminogen activator receptor (uPAR) and active metabolites, but other complex matrices could in all probability be screened with this assay in search of therapeutic compounds of natural origin
In future this system might be coupled to an receptor affinity detection (RAD), enzyme activity/affinity detection (EAD) or on-line antioxidant system to detect the bioactivity of drug metabolites resulting from liver biochips
Summary
Plants have provided the basic building blocks for a number of highly effective drugs and they remain an attractive option for discovery of new molecular entities, due to their still largely untapped chemical diversity [1]. BCD assays can be defined as the detection of bioactives based on biochemical reactions or simulated biochemical reactions. These methods are used to fast-track identification of bioactives in extracts or mixtures without the need for lengthy separation and isolation procedures (see Figure 1 for an example). The power of this type of screening procedure is that it cuts down on in vitro testing, since only new target compounds, showing activity against the specific disease marker, need to be isolated and tested further. In this review the focus will fall on affinity/activity-based BCD methods, including enzyme activity/affinity detection (EAD), receptor affinity detection (RAD), metabolite profiling systems, and antioxidant activity assays, as tools to identify bioactives in plant extracts amongst others
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