Back to Search
Start Over
Detection of Enzyme Inhibitors in Crude Natural Extracts Using Droplet-Based Microfluidics Coupled to HPLC.
- Source :
-
Analytical chemistry [Anal Chem] 2017 May 02; Vol. 89 (9), pp. 4889-4896. Date of Electronic Publication: 2017 Apr 17. - Publication Year :
- 2017
-
Abstract
- Natural product screening for new bioactive compounds can greatly benefit from low reagents consumption and high throughput capacity of droplet-based microfluidic systems. However, the creation of large droplet libraries in which each droplet carries a different compound is a challenging task. A possible solution is to use an HPLC coupled to a droplet generating microfluidic device to sequentially encapsulate the eluting compounds. In this work we demonstrate the feasibility of carrying out enzyme inhibiting assays inside nanoliter droplets with the different components of a natural crude extract after being separated by a coupled HPLC column. In the droplet formation zone, the eluted components are mixed with an enzyme and a fluorogenic substrate that permits to follow the enzymatic reaction in the presence of each chromatographic peak and identify those inhibiting the enzyme activity. Using a fractal shape channel design and automated image analysis, we were able to identify inhibitors of Clostridium perfringens neuraminidase present in a root extract of the Pelargonium sidoides plant. This work demonstrates the feasibility of bioprofiling a natural crude extract after being separated in HPLC using microfluidic droplets online and represents an advance in the miniaturization of natural products screening.
- Subjects :
- Biological Products chemistry
Clostridium perfringens enzymology
Enzyme Assays
Enzyme Inhibitors chemistry
Pelargonium chemistry
Plant Extracts chemistry
Plant Roots chemistry
Zanamivir analysis
Zanamivir chemistry
Biological Products analysis
Chromatography, High Pressure Liquid methods
Enzyme Inhibitors analysis
Microfluidic Analytical Techniques methods
Neuraminidase antagonists & inhibitors
Plant Extracts analysis
Subjects
Details
- Language :
- English
- ISSN :
- 1520-6882
- Volume :
- 89
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- Analytical chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 28374582
- Full Text :
- https://doi.org/10.1021/acs.analchem.6b04988