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Planta Med 2009; 75(3): 280-285
DOI: 10.1055/s-0028-1112195
Analytical Studies
Original Paper
© Georg Thieme Verlag KG Stuttgart · New York

Development and Validation of an RP-HPLC Method for Quantification of Cinnamic Acid Derivatives and Kaurane-Type Diterpenes in Mikania laevigata and Mikania glomerata

Suzan Kelly Vilela Bertolucci1 , 2 , Ana Bárbara Dias Pereira1 , José Eduardo Brasil Pereira Pinto2 , José Antônio de Aquino Ribeiro1 , Alaíde Braga de Oliveira1 , Fernão Castro Braga1
  • 1Department of Pharmaceutical Products, Faculty of Pharmacy, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil
  • 2Agriculture Department, Universidade Federal de Lavras, Lavras, MG, Brazil
Further Information

Publication History

Received: June 4, 2008 Revised: September 26, 2008

Accepted: October 27, 2008

Publication Date:
18 December 2008 (online)

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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 °C). The standard curves were linear over the range 0.5 – 5.0 μg (CO), 0.25 – 4.0 μg (OC), 1.0 – 8.0 μg (BA), 0.5 – 3.0 μg (CA) and 0.8 – 12.0 μg (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 μg and 0.007 – 0.240 μg. The method was tested for new and old columns, temperature variation (26 and 28 °C) and by different operators in the same laboratory. The method was successfully applied to samples of both species.

Key words

Mikania laevigata - Mikania glomerata - Asteraceae - cinnamic acid derivatives - kaurane-type diterpenes - HPLC quantification