Medicinal Chemistry Research

, Volume 26, Issue 12, pp 3315–3322 | Cite as

Rapid screening, separation, and detection of α-glucosidase inhibitors from Hedyotis diffusa by ultrafiltration–liquid chromatography tandem mass spectrometry–high-speed countercurrent chromatography

  • Xiaojing Yang
  • Lina Chen
  • Chunming Liu
  • Yao Qin
  • Ying Tang
  • Senlin Li
Original Research


Hedyotis diffusa is known for its significant medical value, such as curing pneumonia in children, remedying gastritis, and relieving gynecological inflammation. In this study, a new assay based on ultrafiltration–liquid chromatography was developed for the rapid screening and identifying of ligands for α-glucosidase inhibitors from Hedyotis diffusa. Eleven potential α-glucosidase inhibitors were found in the crude extract, and four of these were separated successfully by high-speed countercurrent chromatography: 2-hydroxyl-3-methyl anthraquinone, 2-methyl-3-hydroxyl anthraquinone, 2-hydroxyl-3-methyl-1-methoxyl anthraquinone, and a novel compound. The percent purities of the compounds are 97.71, 95.96, 87.60, and 93.66%, respectively. Results of our study demonstrated that our systematic screening, identification, and isolation of bioactive components in Hedyotis diffusa using ultrafiltration–liquid chromatography–diode array detector, liquid chromatography–electrospray ionization–tandem mass spectrometry, and high-speed countercurrent chromatography are feasible and efficient. Our approach also can be used to rapidly screen and separate other enzyme inhibitors.


Hedyotis diffusa α-glucosidase Ultrafiltration HPLC-PDA-ESI/MS HSCCC 



This work was supported by the National Natural Science Foundation of China [No. 31170326, 31370374]; team project of Jilin Provincial Science and Technology Department [No. 20130413043 GH] and Natural Science Foundation of Changchun Normal University.

Conflict of Interest

The authors declare that they have no competing interests.


  1. Al-Achi A (1998) Greenwood R. Erythrocytes as oral delivery systems for human insulin. Drug Dev Ind Pharm 24:67–72CrossRefPubMedGoogle Scholar
  2. Ganbold M, Barker J, Ma R, Jones L, Carew M (2010) Cytotoxicity and bioavailability studies on a decoction of Oldenlandia diffusa and its fractions separated by HPLC. Ethnopharmacol 131:396–403CrossRefGoogle Scholar
  3. Gao XP, Zhang WY, Zou WJ, Su Z (2003) Screening for α-glucosidase inhibitors from extracts of traditional Chinese medicine. Nat Prod Res Dev 16:36–538Google Scholar
  4. Ghisalberti EL (1998) Phytomedicine 5:147–163CrossRefPubMedGoogle Scholar
  5. Gong J, Xu L (2011) Sequential ASE extraction of alkylphenols from sediments: occurrence and environmental implications. J Hazard Mater 192:643–650CrossRefPubMedGoogle Scholar
  6. Ito Y (2003) Golden rules and pitfalls in selecting optimum conditions for hige-speed counter-current chromatography. J Chromatogr A 1065:145–168CrossRefGoogle Scholar
  7. Kammerer D, Carle R, Schieber A (2004) Characterization of phenolic acids in blackcarrots (Daucus carota ssp. sativus var. atrorubens Alef.) by high-performance liquid chromatography/electrospray ionization mass spectromety, Rapid Commun. J Mass Spectrom 18:1331–1340Google Scholar
  8. Kim DH, Lee HJ, Oh YJ, Kim MJ, Kim SH, Jeong TS, Baek NI (2005) Arch Pharm Res 28:1156–1160CrossRefPubMedGoogle Scholar
  9. Li CM, Xue XY, Zhou DY, Zhang FF (2008) Analysis of iridoid glucosides in Hedyotis diffusa by high-performance liquid chromatography/electrospray ionization tandem mass spectrometry. J Pharmaceut Biomed 48:205–211CrossRefGoogle Scholar
  10. Li H, Song F, Xing J, Tsao R, Liu Z (2009) Screening and structural characterization of α-glucosidase inhibitors from hawthorn leaf flavonoids extract by ultrafiltration LC-DAD-MS n and SORI-CID FTICR MS. J Am Soc Mass Spectr 20(8):1496–1503CrossRefGoogle Scholar
  11. Li SN, Liu CM, Zhang YC (2014) Ultrafiltration liquid chromatography combined with high-speed countercurrent chromatography for screening and isolating potential α-glucosidase and xanthine oxidase inhibitors from Cortex Phellodendri. J Sep Sci 37(18):2504–2512CrossRefPubMedGoogle Scholar
  12. Liu Y, Zhou H, Liu S, Liu Z (2013) Screening of α-glucosidase inhibitors in ginkgo biloba Extract using ultrafiltration LC-ESI-MSn. Chem Res Chin Univ 34(4):813–818Google Scholar
  13. Lu DS, Yong KL, Chen X (2005) Extraction and separation of effective components of guangxi dragon blood of inhibiting α-glucosidase activity. J Shanghai Univ 11:71–73Google Scholar
  14. Perez GRM, Perez GC, Perez GS, Zavaia SMA (1998) Effect of triterpenoids of bouvardia terniflora on bllod sugar levels of normal and alloxan diabetic mice. Phytomedicine 5(6):475–478CrossRefGoogle Scholar
  15. Van de Laar FA, Lucassen PL, Akkermans RP, Van de Lisdonk EH, Rutten GE, Van Weel C (2005) Alpha-glucosidase inhibitors for type 2 diabetes mellitus. Cochrane Database Syst Rev 18:CD003639Google Scholar
  16. Wei MC, Yang YC (2014) Extraction characteristics and kinetic studies of oleanolic and ursolic acids from Hedyotis diffusa under ultrasound-assisted extraction conditions. Sep Sci Technol 130:182–192Google Scholar
  17. Xu GH, Kim YH, Chi SW, Choo SJ, Ryoo IJ, Ahn JS, Yoo ID (2010) Evaluation of human neutrophil elastase inhibitory effect of iridoid glycosides from Hedyotis diffusa. Bioorg Med Chem Lett 20:513–515CrossRefPubMedGoogle Scholar
  18. Yamasaki Y, Katakami N, Hayaishi-Okano R, Matsuhisa M, Kajimoto Y, Kosugi K, Hatano M, Hori M (2005) Alpha-glucosidase inhibitor reduces the progression of carotid intima-media thickness. Diabetes Res Clin Pract 67:204–210CrossRefPubMedGoogle Scholar
  19. Yang JR, Luo JG, Kong LY (2015) Determination of α-glucosidase inhibitors from ScutScutellaria baicalensis using liquid chromatography with quadrupole time of flight tandem mass spectrometry coupled with centrifugal ultrafiltration. Chin J Nat Med 13(3):0208–0214Google Scholar
  20. Yoshikuni Y (1998) Inhibition of intestinal α-glucosidase activity and postpran dialhy- peiglycemia by moranoline and its N-alkyl Derivative. Agric Biol Chem 52:121–128Google Scholar
  21. Zhang JZ (1995) The diagnosis and features of various types of diabetes. Chin Pract J Rural Dr 11:10–11Google Scholar
  22. Zhao SY, Huang WL (1999) The research progress of diabetes drugs. North Am Clin Drug 20:130–135Google Scholar
  23. Zhou H, Xing JP, Liu S (2012) Screening and Determination for potential a-glucosidase inhibitors from leaves of Acanthopanax senticosus Harms by Using UF-LC/MS and ESI-MSn. Phytochem Lett 24:315–323CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • Xiaojing Yang
    • 1
  • Lina Chen
    • 1
  • Chunming Liu
    • 1
  • Yao Qin
    • 1
  • Ying Tang
    • 1
  • Senlin Li
    • 1
  1. 1.Central LaboratoryChangchun Normal UniversityChangchunChina

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