The Determination of the Iridoids from the Melampyrum species by Modern Chromatographic Methods

Abstract

The idea of studying the iridoids from the Melampyrum bihariense Kern has started from the few data found about the chemical composition of this species. The literature mentions the presence of iridoids found in two species of Melampyrum: Melampyrum arvense and Melampyrum cristatum. The study consists of a qualitative and quantitative analysis by modern chromatographic methods, TLC.TLC coupled with photo-densitometry as well as chromatography on the high performance column (LC/MS). The methanol extract was done from the flowers and leaves of Melamphyrum bihariense. For the TLC and HPLC, the aucubin was used as standard solution. The obtained chromatogram was examined in visible light spectrum, and then it was revealed with sulphuric anisaldehyde, emphasizing the brown coloured spots. In the TLC case, the chromatographic plate was scanned with a Shimadzu CS9000 photo densitometer after spraying with iron chloride, and for the HPLC the analytical column Atlantis HILIC was used 100 mm x 3.0 mm, 3.5 ?m (water), and the ESI source of ions (electrospray ionisation). After the TLC analysis, the iridoids concentration found in the aucubin is 0.23 mg/mL in flowers and 0.24 mg/mL in leaves, and by the LC/MS method the concentration is of 1.8125 mg/ml in flowers and 2.3037 mg/ml in leaves.