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Journal of Analytical Methods in Chemistry
Volume 2017 (2017), Article ID 1963908, 10 pages
Research Article

SPE-UHPLC-FLD Method for the Simultaneous Determination of Five Anthraquinones in Human Urine Using Mixed-Mode Bis(tetraoxacalix[2]arene[2]triazine) Modified Silica as Sorbent

1Henan University of Chinese Medicine, Zhengzhou 450008, China
2College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou 450052, China

Correspondence should be addressed to Kai Hu

Received 10 March 2017; Revised 13 July 2017; Accepted 20 August 2017; Published 28 September 2017

Academic Editor: Hassan Y. Aboul Enein

Copyright © 2017 Kai Hu et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


The five anthraquinones compounds (including aloe-emodin, emodin, physcion, chrysophanol, and rhein) are regarded as the main effective ingredients in rhubarb (Dahuang in Chinese, one of the commonly used Chinese herbal medicines). In this work, a simple and effective solid phase extraction (SPE) method based on bis(tetraoxacalix[2]arene[2]triazine) modified silica gel as adsorbent was developed. Coupled with UHPLC-FLD, the developed method was successfully applied for the measuring of main anthraquinones in human urine after oral administration of the extracts of rhubarb. To obtain the highest recoveries of the five anthraquinones in the SPE process, the main parameters which may affect extraction efficiency were optimized. The optimized sorbent amount, sample loading pH, sample loading rate, washing solution, and eluent condition were obtained. The developed method showed good linearity in 0.012–1.800 μg mL−1 for the five anthraquinones with correlation coefficients more than 0.9993. The investigated LOD values ranged from 3.9 to 5.7 ng mL−1, while the LOQs were between 12.0 and 18.2 ng mL−1. The recoveries of the method were also investigated, which were in the range of 94.8–106.6%. The application of the mixed-mode SPE materials in the proposed method was feasible and simple, and suitable for the enrichment of anthraquinones in urine samples.