Quantitative determination of anti-fungal and insecticide amides in adult plants, plantlets and callus fromPiper tuberculatumby reverse-phase high-performance liquid chromatography (original) (raw)

2003, Phytochemical Analysis

GC-MS has also proved to be a viable technique to quantify pyridone amides (Dodson et al., 2000). We have previously isolated and determined the structures of the amides dihydropiplartine (1), piplartine (2), ∆ α,β-dihydropiperine (3) and pellitorine (4) from mature Piper tuberculatum plants (Navickiene et al., 2000). The isolated compounds have now been used as standards in the quantitative analysis of these bioactive amides by HPLC with photodiode array detection (PAD) in plantlets in vitro and ex vitro, and in callus of P. tuberculatum. EXPERIMENTAL Materials. HPLC grade methanol was purchased from Mallinckrodt Baker (Xalostoc, México); nanopure water (> 18 MΩ) was obtained using a Millipore (Bedford, MA, USA) purifier. All solvents and samples were filtered through 0.2 µM nylon membranes prior to use. Compounds 1-4 were isolated from seeds and leaves of P. tuberculatum as described previously (Navickiene et al., 2000). The NMR, IR and MS data of these compounds matched those previously reported (