Quantitative HPLC Analysis of Deoxyshikonin, Acetyl shikonin and 3-Hydroxy-isovaleryl shikonin in Onosma armeniacum root

Onosma armeniacum Klokov (Boraginaceae) has been used for wounds, burns and hemorrhoids and to treat stomach ulcer and tonsillitis in Turkey [1]. It is known as „havacıva“ in Erzurum Province (Turkey). The roots of some Onosma species contain naphthoquinone compounds [2]. In this study, the quantitative analysis of deoxyshikonin (1), acetyl shikonin (2) and 3-hydroxy-isovaleryl shikonin (3) isolated from n-hexane-dichloromethane (1:1) extract of O. armeniacum roots was carried out. HPLC analysis was performed on a Thermoquest HPLC system, equipped with a DAD detector (Thermo UV 6000). For all analysis, a RP-C18 column (250×4.6mm, 5µm particle size, ACE®) was used. The mobile phase consisted of acetonitril-methanol-water (2% acetic acid) (60:20:20), which were applied in the isocratic elution. The analysis temperature was kept constant at ambient temperature, flow rate and sample volume were set to 1.0ml/min and 20µl, respectively. All analysis were monitored at 525 nm. Peaks were assigned by spiking the samples with authentic samples of 1, 2 and 3, and comparison of the UV-spectra and retention times. Relative contents of the acetyl shikonin, deoxyshikonin and 3-hydroxy-isovaleryl shikonin in the extracts were 23.82%, 3.63% and 2.53%, respectively.

References: [1] Özgen U., Coşkun M. (2000) Ilıca (Erzurum) İlçesine Bağlı Köylerde Halk İlacı Olarak Kullanılan Bitkiler, XIII. Bitkisel İlaç Hammaddeleri Toplantısı, 20–22 September, İstanbul (in Turkish). [2] Khajuria, R.K., Jain, S.M. (1993). Indian J. Chem. B 32, 390–391.