Simultaneous determination of eight anthraquinones in Semen Cassiae by HPLC-DAD

Lijia Xu, Chi On Chan, Ching Ching Lau, Zhiling YU, Daniel K.W. Mok*, Sibao Chen

*Corresponding author for this work

Research output: Contribution to journalJournal articlepeer-review

49 Citations (Scopus)


Introduction Semen Cassiae (SC), a traditional Chinese herbal medicine for the treatment of various diseases, is known to contain active anthraquinone ingredients. However, since the content of some anthraquinones is too low, previous analytical methods only allow the quantitation of a few anthraquinones or a hydrolysis step has to be included in the sample preparation. A rapid and accurate method to examine the content of as many anthraquinones as possible in SC would be desirable. Objective To develop a rapid, sensitive and accurate high-performance liquid chromatography-diode array detection (HPLC- DAD) method to simultaneously quantify eight major anthraquinones (obtusifolin-2-glucoside, aurantio-obtusin, aloe- emodin, rhein, obtusifolin, emodin, chrysophanol and physcion) in SC. Methodology The separation of anthraquinones was achieved on a C18-column with a gradient elution using acetonitrile and 0.1% phosphoric acid. The detection wavelength was 278 nm and the analysis finished within 25 min. Results The limits of detection of these compounds ranged from 0.07 to 0.15 μg/mL while the limits of quantitation ranged from 0.24 to 0.51 μg/mL. All calibration curves showed good linearities (r2 > 0.999) within the test ranges. This validated method was successfully used to analyse 22 batches of SC samples collected from various geographical locations. Conclusion The method was validated to be simple, rapid, accurate and reliable to simultaneously determine eight major anthraquinones in SC. Meanwhile, a more specific anthraquinone, obtusifolin, was proposed to serve as a marker for SC, replacing chrysophanol.

Original languageEnglish
Pages (from-to)110-116
Number of pages7
JournalPhytochemical Analysis
Issue number2
Publication statusPublished - 2012

Scopus Subject Areas

  • Analytical Chemistry
  • Food Science
  • Biochemistry
  • Molecular Medicine
  • Plant Science
  • Drug Discovery
  • Complementary and alternative medicine

User-Defined Keywords

  • anthraquinone
  • Cassia obtusifolia L.
  • obtusifolin
  • Semen Cassiae


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