Chemical Fingerprinting by HPLC-DAD to Differentiate Certain Subspecies of Origanum vulgare L.
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Chemical Fingerprinting by HPLC-DAD to Differentiate Certain Subspecies of Origanum vulgare L. María Dolores González 1 & Paula Laura Lanzelotti 1 & Cecilia María Luis 1
Received: 21 July 2016 / Accepted: 19 October 2016 # Springer Science+Business Media New York 2016
Abstract Origanum vulgare L. is one of the most important herbs in world trade. Because this is a polymorphic species with several subspecies and hybrids, the identification of populations based on morphological features is sometimes difficult. With the aim to find reference profiles, in the present study, we developed a high-performance liquid chromatography method with diode array detection (HPLC-DAD) analysis to describe the chromatographic profile of the hydroalcoholic extract of a certified sample of O. vulgare ssp. vulgare. In agreement with that found in our phytochemical studies, we identified main polyphenol peaks, showing different polyphenolic composition in the O. vulgare L. subspecies vulgare, virens, and hirtum. These differences between subspecies were due to the absence/presence or abundance of closely related structures derived from the benzyl alcohol 4-O-glucosides gastrodin and calleryanin. These results were also evident in thin layer chromatography. The compounds of interest for differentiation were separated using reverse phase with gradient elution, at a wavelength of 280 nm. Discrimination of subspecies vulgare and virens was achieved by comparing their HPLC fingerprint chromatograms using their typical marker peaks while these compounds were absent in subspecies hirtum. Total phenol concentration, DPPH antioxidant potential, and average concentration of esters of gastrodin and calleryanin in the different subspecies were also comparatively determined. These results provide a methodology to
* María Dolores González [email protected]
1
Basic Sciences Department, Universidad Nacional de Luján, Cruce Av. Carlos Pellegrini y Ruta 7, 6700 Luján, Provincia de Buenos Aires, Argentina
differentiate those O. vulgare L. subspecies for good agricultural practices and selection. Keywords Origanum vulgare ssp. vulgare . HPLC-DAD fingerprinting . Gastrodin 3,4-dihydroxybenzoate . Calleryanin 3,4-dihydroxybenzoate . Oregano subspecies differentiation
Introduction Origanum is a complex genus of the family Lamiaceae, reviewed by Ietswaart (1980). The most abundant species of this genus is Origanum vulgare L. This species, native to the Mediterranean up to the north of Europe and from Western Europe to China, has been introduced worldwide from Europe. In North and South America, it has been grown for many years. O. vulgare L. is a polymorphic species, which has been subdivided in six subspecies (vulgare, viride, virens, gracile, glandulosum, and hirtum) and even some hybrids have been reported (Ietswaart 1980). The identification of populations is difficult, especially in commercial materials that have lost most of their anatomic features. Although O. vulgare L. is a very important culinary herb, consumed since ancient times, it is yet
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