Simultaneous Determination of Eight Bioactive Compounds in Dianthus superbus by High‑performance Liquid Chromatography

Background: Dianthus superbus, one of traditional herbal medicine, is widely used to treat urethritis, carbuncles and carcinoma. Objective: A simultaneous determination method was established for controlling the quality of D. superbus using the eight compounds, (E)‑methyl‑4‑hydroxy‑4‑(8a‑methyl‑3‑oxodecahydronaphthalen‑4a‑yl) (1), diosmetin‑7‑O(2’’,6’’‑di‑O‑α‑L‑rhamnopyranosyl)‑β‑D‑glucopyranoside (2), vanillic acid (3), 4‑hydroxyphenyl acetic acid (4), 4‑methoxyphenyl acetic acid (5), (E)‑4‑methoxycinnamic acid (6), 3‑methoxy‑4‑hydroxyphenylethanol (7), and methyl hydroferulate (8) isolated from D. superbus. Materials and Methods: This analysis method was developed using high performance liquid chromatography coupled with diode array detector with a Shishedo C18 column at a column temperature of 3°C. The mobile phase was composed of 0.1% trifluoroacetic acid in water and acetonitrile. The flow rate was 1 ml/min and detection wavelength was set at 205 nm and 280 nm. Validation was performed in order to demonstrate selectivity, accuracy and precision of the method. Results: The calibration curves showed good linearity (R2 > 0.99). The limits of detection and limits of quantification were within the ranges 0.0159–0.6205 µg/ml and 0.3210–1.8802 µg/ml, respectively. Moreover, the relative standard deviations of intra‑ and inter‑day precision were both <2.98%. The overall recoveries were in the range of 96.23–109.87%. Quantitative analysis of eight compounds in 12 D. superbus samples (D‑1–D‑12) from various regions were analyzed and compared by developed method. Conclusion: As a result, this established method was accurate and sensitive for the quality evaluation of eight compounds isolated from D. superbus and may provide a new basis for quality control of D. superbus.