Each piperaceae extract was prepared at a 200 µg/mL concentration in 95% ethanol (p.a), whereas a standard piperine solution was prepared at 100 µg/mL. Analysis was performed using TLC-densitometry with 60 F254 silica gel TLC plates as a stationary phase and hexane-ethyl acetate (1:1) as a mobile phase. 3 µL sample and standard piperine were spotted, then eluted with mobile phase until the spot appeared and was observed under a UV lamp 254 nm.
Protocol detail
Quantitative Analysis of Piperaceae Extracts
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Variable analysis
independent variables
- Concentration of piperaceae extract (200 µg/mL)
- Concentration of standard piperine solution (100 µg/mL)
- Chromatographic profile (Retention factor, RF) of piperaceae extract and standard piperine
- Solvent used for preparation of extracts and standard solution (95% ethanol)
- Stationary phase (60 F254 silica gel TLC plates)
- Mobile phase (hexane-ethyl acetate, 1:1 ratio)
- Sample volume spotted (3 µL)
- Detection method (UV lamp, 254 nm)
- Standard piperine solution (100 µg/mL)
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