Simultaneous determination of biological marker compounds in ostericum koreanum by HPLC method and discrimination by principal component analysis

An analytical method for the simultaneous determination of four biological marker compounds including bisabolangelone, oxypeucedanin, imperatorin, and isoimperatorin extracted from Ostericum koreanum has been developed by HPLC-UV detection at a wavelength 254 nm. Moreover, biological active compounds have been identified by electrospray ionization tandem mass spectrometry (ESI-MS/MS). In ESI-MS/MS spectra, these compounds produced several characteristic ions through collisional-induced dissociation, enabling to clarify these compounds. HPLC chromatographic separation was successfully achieved with a Develosil RPAQUEOUS C₃₀ (4.6 × 250 mm, 5 μm) column and a mobile phase of acetonitrile-water (60:40, v/v). This developed analytical method was validated with specificity, selectivity, accuracy and precision. The method was deemed satisfactory by inter- and intra-day validation and exhibited both high accuracy and precision (relative standard deviation < 9.2%). Overall limits of quantitation and detection were approximately 0.03-0.05 μg/mL at a signal-to-noise ratio (S/N) of 3 and were about 0.10-0.25 μg/mL at a S/N of 10. In addition, principal component analysis (PCA) was performed on the analytical data of 14 different Ostericum koreanum samples in order to classify samples collected from different regions. This method has been successfully applied to quality control of Ostericum koreanum extracts.