Development of a sensitive LC–MS/MS method for quantification of coniferyl ferulate and its metabolite coniferyl alcohol in rat plasma: Application to a pharmacokinetic study

Highlights

• Coniferyl ferulate and its metabolite coniferyl alcohol were simultaneously determined.

• Method validation procedures were in accordance with EMA regulations.

• It was successfully applied to the pharmacokinetic study in rats.

Abstract

A rapid and simple LC–MS/MS method was developed and validated for the simultaneous determination of coniferyl ferulate (CF) and its metabolite coniferyl alcohol (CA) using bavachromene as an internal standard (IS). A TSQ Quantum Access mass spectrometer was operated under selected-reaction monitoring mode using negative electrospray ionization. Extraction with ether was used in sample preparation. The plasma samples were prepared and then chromatographed on a Phenomenex Luna C₁₈ column (2.1 mm × 50 mm, 1.7 μm; Torrance, USA) at 35 °C, using acetonitrile: water (65:35, v/v) in an isocratic mode at a flow rate of 0.3 mL/min. Method validation was performed as per the FDA guidelines and calibration curves showed good linearity over the concentration range of 2.5–1000 ng/mL for both CF and CA. The intra- and inter-day precision and accuracy were within the acceptable limits. The developed assay was successfully applied to a pharmacokinetic study of CA in rats.

Introduction

Coniferyl ferulate (CF, see Fig. 1) was isolated from Levisticum officinale, Angelica sinensis, and Ligusticum chuanxiong [1], [2]. These herbal medicines have long been used as traditional Chinese medicines for the treatment of female irregular menstruation, amenorrhea rheumatic arthralgia, menstrual disorders, and swelling pain [3], [4]. The compound CF exhibits multiple pharmacological activities such as antibacterial [5], antioxidant [6], anticancer [7], [8], and vasodilating effects [9]. Moreover, CF could significantly inhibit CCl₄-induced hepatic fibrosis in mice, which mediated through inhibiting TGF-β signaling pathway [10]. Clearly, it is of interest to further evaluate the medicinal potential of cf. There are just a few reports that separated and determined CF from Angelica sinensis and Ligusticum chuanxiong by HPLC methods [11], [12], [13], but these methods were not available for the pharmacokinetic application because of low sensitivity and poor selectivity. In this study, we established a rapid and simple LC–MS/MS method for simultaneous determination of CF and its active metabolite coniferyl alcohol (CA) in rat plasma, and it was successfully applied to the pharmacokinetic study of CF.