Determination of Anthraquinones by UPLC Method in Patient with Craniocerebral Injuries after Oral Administration of Rhubarb

Ultra-high performance liquid chromatography (UPLC) method was developed for determining anthraquinones in plasma of patient with craniocerebral injuries after administration of Rhubarb. Chromatographic separation was performed on an Acquity UPLC BEH C₁₈ column (2.1 mm × 100 mm, 1.7 μm) with methanol-0.5% aqueous acetic acid (12:82) as mobile phase at a flow rate of 0.5 mL·min⁻¹. The wavelength was set at 254 nm. The operating temperature was maintained at 25°C and the sample volume injected was 5 μL. Linear range was satisfactory for determination of rhein in human plasma with the concentration ranging from 0.42 to 13.429 μg·mL⁻¹. Four other anthraquinones were not detected using UPLC method. Recovery rate of this method was within the range of 97.2%−103.11%. The relative standard deviation of intra- and inter-day was less than 6%. The concentration of rhein in plasma after 1 hour administration from five TBI patients was 7.93±1.94 μg·mL⁻¹. The UPLC is a method with good precision, high recovery rate, simple operation, innocuity, and good reproducibility.