Development and Validation of an RP-HPLC Method for Quantification of Cinnamic Acid Derivatives and Kaurane-Type Diterpenes inMikania laevigataandMikania glomerata

Abstract

MIKANIA GLOMERATA and MIKANIA LAEVIGATA (Asteraceae) are medicinal plants popularly named 'guaco' in Brazil. The leaves of both species are used to treat respiratory diseases, with coumarin (CO) and kaurane-type diterpenes being regarded as the bioactive constituents. A new and simple RP-HPLC method was developed and validated for the simultaneous quantification of CO, O-coumaric (OC), benzoylgrandifloric (BA), cinnamoylgrandifloric (CA) and kaurenoic (KA) acids in the species. Optimal separation was achieved with an alternating gradient elution of methanol and acetonitrile and detection was carried out by DAD at three different wavelengths: 210 nm for CO, OC, KA; 230 nm for BA; and 270 nm for CA. The extracts showed good stability during 42 hours under normal laboratory conditions (temperature of 23 +/- 2 degrees C). The standard curves were linear over the range 0.5 - 5.0 microg (CO), 0.25 - 4.0 microg (OC), 1.0 - 8.0 microg (BA), 0.5 - 3.0 microg (CA) and 0.8 - 12.0 microg (KA), with R(2) > 0.999 for all compounds. The method showed good precision for intra-day (RSD < 4.6 %) and inter-day assays (RSD < 4.4 %). The recovery was between 99.9 and 105.3 %, except for CO and OC in M. glomerata (73.2 - 91.6 % and 86.3 - 117.4 %, respectively). The limits of quantification and detection were in the range of 0.025 - 0.800 microg and 0.007 - 0.240 microg. The method was tested for new and old columns, temperature variation (26 and 28 degrees C) and by different operators in the same laboratory. The method was successfully applied to samples of both species.

Anti-hemorrhagic effect of hydro-alcoholic extract of the leaves of Mikania glomerata in lesions induced by Bothrops jararaca venom in rats

“…The main chemical compound with medicinal activity, which has been reported to exert antiophidic activity 16 , is coumarin 17,18 .…”

Efficacy of guaco mouthwashes (Mikania glomerata and Mikania laevigata) on the disinfection of toothbrushes

“…Coumarin was detected in samples of M. glomerata (Table 2), but in concentrations bellow the limit of quantification established for the method. The presence of coumarin in samples of M. glomerata has been reported in several articles (Castro et al, 2006), as well as its absence (Bertolucci et al, 2009).…”

Isolation and HPLC quantitation of kaurane-type diterpenes and cinnamic acid derivatives of long-term stored leaves of Mikania laevigata and Mikania glomerata

“…Previous reports have suggested similar chemical composition for the leaves of both Mikania (Veneziani and Oliveira 1999, Vilegas et al Mikania (Veneziani and Oliveira 1999, Vilegas et al Mikania 1997, Oliveira et al 1984. However, we reported a marked difference in the constitution of M. laevigata and M. glomerata leaves, with the presence of KA, BA and CA in both and the lack of CO and OC in the last one (Bertolucci et al 2009). Besides, we found higher contents of BA and CA in M. laevigata, while KA was disclosed as the most abundant compound in M. glomerata (Bertolucci et al 2009).…”

“…The validation of medicinal plants has only gained attention of the Brazilian scientifi c community and government in the last decade (Bertolucci et al 2009); for that reason, the quality control of herbal drugs in the country is limited by the reduced number of chemical markers defi ned for native species, along with the lack of commercial sources of reference compounds (Braga et al 2003).…”