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Journal of Integrative Medicine ›› 2014, Vol. 12 ›› Issue (5): 455-462.doi: 10.1016/S2095-4964(14)60037-6

• Research Article • Previous Articles     Next Articles

Simultaneous determination of five anthraquinones in a Chinese traditional preparation by RP-HPLC using an improved extraction procedure

Yan-bin Shia, Hui-li Lia, Hai-qin Wanga, Yan-biao Yangb, Xiao-yun Zhanga, Hui Wanga, Zong-jie Zhub, Zhi-ye Zhangb, Cheng-an Zhangb   

  1. School of Pharmacy, Lanzhou University, Lanzhou 730000, Gansu Province, China
    Department of Pharmaceutics, the First People’s Hospital of Lanzhou, Lanzhou 730000, Gansu Province, China
  • Received:2014-02-17 Accepted:2014-05-07 Online:2014-09-10 Published:2014-09-15


The stable quality of Chinese herbal medicines is a critical factor for their reliable clinical efficiency. An improved liquid-liquid extraction procedure and a liquid chromatographic method were developed to simultaneously analyze five anthraquinones (aloe-emodin, rhein, emodin, chrysophanol and physcion) in a Chinese traditional hospital preparation, Fuyankang mixture, in order to quantitatively control its quality in a more effective way. 


A more economical and repeatable extraction procedure based on conventional liquid-liquid extraction technique was developed and used to extract five marker components in Fuyankang mixture. These anthraquinones were separated in less than 20 min on a C18 column with methanol and 0.1% phosphoric acid (88:12, v/v) as mobile phase. The method was validated for specificity, precision, spiked recovery and stability. 


Compared to conventional liquid-liquid extraction, the improved liquid-liquid extraction was found to be more effective for simultaneous extraction of anthraquinones from an aqueous Chinese herbal preparation, especially for hydrophobic compounds. The improved extraction method was successfully applied to determine the content of five marker components in Fuyankang mixture by the means of reverse phase high-performance liquid chromatography. 


The improved extraction procedure may be suitable for routine quality control of Fuyankang mixture and other traditional preparations at city-level hospitals in China.

Key words: Improved liquid-liquid extraction, Fuyankang mixture, Drugs, Chinese herbal, Chromatography, High-performance liquid, Anthraquinones, Quality control

Figure 1

Chemical structure of analytes[8]Thus, the aim of the present investigation was to develop a convenient, effective, and reproducible extraction method based on conventional LLE technique for the analysis of anthraquinones in Fuyankang mixture using RP-HPLC. This method is necessary to establish routine quality control for this preparation or other TCM preparations at city-level hospitals in China."

Figure 2

The equipment of an improved liquid-liquid extraction"

Figure 3

Chromatograms of the five analytesA: Mixed standard solution (all concentrations of compounds 1 to 5 were 2.5 μg/mL); B: Fuyankang mixture sample spiked with reference substances (30 μg); C: Fuyankang mixture sample (Lot No. 20120207). Peaks: 1=aloe-emodin, 2=rhein, 3=emodin, 4=chrysophanol, 5=physcion. Analytical conditions: column, Diomonsil? C18; mobile phase, methanol-0.1% phosphoric acid (88:12, v/v); flow-rate, 0.8 mL/min; column temperature, 40 ℃; detection wavelength, 254 nm."





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