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Determination of Triterpene Acids from 37 Different Varieties of Raspberry using Pre-column Derivatization and HPLC Fluorescence Detection

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Abstract

In this work, we have developed an efficient method for the rapid extraction and separation of triterpene acids from 37 different varieties of raspberry via ultrasound-assisted dispersive liquid–liquid microextraction (UA-DLLME). The triterpene acids were then determined by high-performance liquid chromatography (HPLC) with fluorescence detection using benzimidazo-[2,1-b]quinazolin-12(6H)-one-5-ethyl-p-toluenesulfonate (BQETS) as the labeling agent. Five triterpene acids, including asiatic acid (AA), maslinic acid (MA), corosolic acid (CA), oleanolic acid (OA) and betulinic acid (BA), were extracted by UA-DLLME using chloroform and acetone as the extracting and dispersing solvents, respectively. After extraction and nitrogen flushing, the extracts were simultaneously characterized by HPLC based on pre-column derivatization using BQETS, a new labeling agent synthesized in our laboratory. Several key parameters affecting the extraction efficiency and derivatization yields were investigated and optimized by response surface methodology (RSM) combined with Box–Behnken design (BBD). The method was further validated for linearity (correlation coefficient R 2 > 0.9979), precision (RSD = 0.23–2.45 %), and recovery (RSD = 90–106.5 %). The limits of detection (LODs) and the limits of quantification (LOQs) were determined to be within the range of 1.83–7.69 µg/L and 6.06–25.47 µg/L, respectively. This is the first report of the use of BQETS as a pre-column derivatization agent for the determination of triterpene acids in real samples. The proposed method has been applied to the determination of five triterpene acids in 37 different raspberry varieties with significantly increased sensitivity compared to other methods. The results obtained indicate that the contents of triterpene acids vary significantly across different raspberry varieties.

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References

  1. Chen Y, Wang MF, Rosen RT et al (1999) 2, 2-Diphenyl-1-picrylhydrazyl radical-scavenging active components from Polygonum multiflorum Thunb. J Agr Food Chem 47(6):2226–2228

    Article  CAS  Google Scholar 

  2. Četojević-Simin DD, Velićanski AS, Cvetković DD et al (2015) Bioactivity of Meeker and Willamette raspberry (Rubus idaeus L.) pomace extracts. Food Chem 166:407–413

    Article  Google Scholar 

  3. Gonzalez-Barrio R, Edwards CA, Crozier A (2011) Colonic catabolism of ellagitannins, ellagic acid, and raspberry anthocyanins. In vivo and in vitro studies. Drug Metab Dispos 39:1680–1688

    Article  CAS  Google Scholar 

  4. Verbeyst L, Crombruggen KV, Plancken IV, Hendrickx Marc, Loey AV (2011) Anthocyanin degradation kinetics during thermal and high pressure treatments of raspberries. J Food Eng 105:513–521

    Article  CAS  Google Scholar 

  5. Bobinaite B, Pranas Viškelis P, Venskutonis PR (2012) Variation of total phenolics, anthocyanins, ellagic acid and radical scavenging capacity in various raspberry (Rubus spp.) cultivars. Food Chem 132:1495–1501

    Article  CAS  Google Scholar 

  6. Dossetta M, Leeb J, Finn CE (2010) Variation in anthocyanins and total phenolics of blackraspberry populations. J Funct Foods 2:292–297

    Article  Google Scholar 

  7. Sariburun S, Şahin S, Demir C et al (2010) Phenolic content and antioxidant activity of raspberry and blackberry cultivars. J Food Sci 75:C328–C335

    Article  CAS  Google Scholar 

  8. Mullen W, Stewart AJ, Lean MEJ et al (2002) Effect of freezing and storage on the phenolics, ellagitannins, flavonoids, and antioxidant capacity of red raspberries. J Agric Food Chem 50:5197–5201

    Article  CAS  Google Scholar 

  9. Mullen W, McGinn J, Lean ME et al (2002) Ellagitannins, flavonoids, and other phenolics in red raspberries and their contribution to antioxidant capacity and vasorelaxation properties. J Agric Food Chem 50(18):5191–5196

    Article  CAS  Google Scholar 

  10. Gbaguidi F, Accrombessi G, Moudachirou M et al (2005) HPLC quantification of two isomeric triterpenic acids isolated from Mitracarpus scaber and antimicrobial activity on Dermatophilus congolensis. J Pharm Biomed Anal 39:990–995

    Article  CAS  Google Scholar 

  11. Novotny L, Abdel-Hamid ME, Hamza H et al (2003) Development of LC–MS method for determination of ursolic acid: application to the analysis of ursolic acid in Staphylea holocarpa Hemsl. J Pharm Biomed Anal 31:961–968

    Article  CAS  Google Scholar 

  12. Gupta A, Sheth NR, Pandey S et al (2013) Determination of ursolic acid in fractionated leaf extracts of Ocimum gratissimum Linn and in developed herbal hepatoprotective tablet by HPTLC. Pharmacogn J 5:156–162

    Article  Google Scholar 

  13. Yu DL, Sakurai Y, Chen CH et al (2006) Anti-AIDS agents 69. Moronic acid and other triterpene derivatives as novel potent anti-HIV agents. J Med Chem 49:5462–5469

    Article  CAS  Google Scholar 

  14. Wójciak-Kosior M (2007) Separation and determination of closely related triterpenic acids by high performance thin-layer chromatography after iodine derivatization. J Pharm Biomed Anal 45:337–340

    Article  Google Scholar 

  15. Kontogianni VG, Exarchou V, Troganis A et al (2009) Rapid and novel discrimination and quantification of oleanolic and ursolic acids in complex plant extracts using two-dimensional nuclear magnetic resonance spectroscopy–comparison with hplc methods. Ana Chim Acta 635(2):188–195

    Article  CAS  Google Scholar 

  16. Guo S, Duan JA, Tang YP et al (2010) Characterization of triterpenic acids in fruits of Ziziphus species by HPLC–ELSD–MS. J Agric Food Chem 58(10):6285–6289

    Article  CAS  Google Scholar 

  17. Yuan Y, Zhang H, Sun F et al (2015) Biopharmaceutical and pharmacokinetic characterization of asiatic acid in Centella asiatica as determined by a sensitive and robust HPLC-MS method. J Ethnopharmacol 163:31–38

    Article  CAS  Google Scholar 

  18. Yang P, Li Y, Liu X et al (2007) Determination of free isomeric oleanolic acid and ursolic acid in Pterocephalus hookeri by capillary zone electrophoresis. J Pharm Biomed Anal 43(4):1331–1334

    Article  CAS  Google Scholar 

  19. Banerjee A, Sane RT, Mangaonkar K et al (2006) Quantitation of oleanolic acid in Oldenlandia corymbosa L. whole-plant powder by high-performance thin-layer chromatography. JPC-J Planar Chromatogr-Mod TLC 19:68–72

    Article  CAS  Google Scholar 

  20. Banno N, Akihisa T, Tokuda H et al (2004) Triterpene acids from the leaves of Perilla frutescens and their anti-inflammatory and antitumor-promoting effect. Biosci Biotechnol Biochem 68:85–90

    Article  CAS  Google Scholar 

  21. Werner S, Nebojsa S, Robert W et al (2003) Complete assignments of 1H and 13C NMR resonances of oleanolic acid, 18α-oleanolic acid, ursolic acid and their 11-oxo derivatives. Magn Reson Chem 41(8):636–638

    Article  Google Scholar 

  22. Caligiani A, Malavasi G, Palla G et al (2013) A simple GC–MS method for the screening of betulinic, corosolic, maslinic, oleanolic and ursolicacid contents in commercial botanicals used as food supplement ingredients. Food Chem 136(2):735–741

    Article  CAS  Google Scholar 

  23. Kartini Piyaviriyakul S, Siripong P et al (2014) HPTLC simultaneous quantification of triterpene acids for quality control of Plantago major L. and evaluation of their cytotoxic and antioxidant activities. Ind Crops Prod 60:239–246

    Article  CAS  Google Scholar 

  24. Lv T, Zhao XN, Zhu SY et al (2014) Development of an efficient HPLC fluorescence detection method for brassinolide by ultrasonic assisted dispersive liquid–liquid microextraction coupled with derivatization. Chromatographia 77:1653–1660

    Article  CAS  Google Scholar 

  25. Sun YN, Zhang X, Ji ZY et al (2016) Determination of free fatty acids of Chinese Coriandrum sativum L. Using Benzimidazo[2, 1-b] quinazolin-12 (6H)-one-5-ethyl-p-toluenesulfonate as precolumn labeling reagent by LC with fluorescence detection. Chromatographia 79:547–559

    Article  CAS  Google Scholar 

  26. Song M, Hang TJ, Wang Y et al (2006) Determination of oleanolic acid in human plasma and study of its pharmacokinetics in Chinese healthy male volunteers by HPLC tandem mass spectrometry. J Pharm Biomed Anal 40:190–196

    Article  CAS  Google Scholar 

  27. Claude B, Morin P, Lafosse M et al (2008) Selective solid-phase extraction of a triterpene acid from a plant extract by molecularly imprinted polymer. Talanta 75:344–350

    Article  CAS  Google Scholar 

  28. Moradi M, Yamini Y, Seidi S et al (2013) Ultrasound-assisted emulsification microextraction using low density solvent for analysis of toxic nitrophenols in natural waters. Int J Environ Anal Chem 93:199–212

    Article  CAS  Google Scholar 

  29. Vas G, Vekey K (2004) Solid-phase microextraction: a powerful sample preparation tool prior to mass spectrometric analysis. J Mass Spectrom 39:233–254

    Article  CAS  Google Scholar 

  30. Yan HY, Wang H (2013) Recent development and applications of dispersive liquid–liquid microextraction. J Chromatogr A 1295:1–15

    Article  CAS  Google Scholar 

  31. Zhao XN, Lv T, Zhu SY et al (2016) Dual ultrasonic-assisted dispersive liquid–liquid microextraction coupled with microwave-assisted derivatization for simultaneous determination of 20 (S)-protopanaxadiol and 20 (S)-protopanaxatriol by ultrahigh performance liquid chromatography-tandem mass spectrometry. J Chromatogr A 1437:49–57

    Article  CAS  Google Scholar 

  32. Leong MI, Fuh MR, Huang SD (2014) Beyond dispersive liquid–liquid microextraction. J Chromatogr A 1335:2–14

    Article  CAS  Google Scholar 

  33. Viñas P, Campillo N, López-García I et al (2014) Dispersive liquid–liquid microextraction in food analysis. A critical review. Anal Bioanal Chem 406(8):2067–2099

    Article  Google Scholar 

  34. Yeh WC (2003) A MCS-RSM approach for network reliability to minimise the total cost. Int J Adv Manuf Technol 22(9–10):681–688

    Article  Google Scholar 

  35. Ebrahimi-Najafabadi H, Leardi R, Jalali-Heravi M (2014) Experimental design in analytical chemistry—part I: theory. J AOAC Int 97(1):3–11

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors acknowledge the financial support from major science and technology projects of Qinghai Province: utilization and development with high value and further process technology of medlar (2015-NK-A2), and the unit of science and technology office of Qinghai province: manufacture integration technology and demonstration of high standards medlar with antioxidant and anti-cohesion (2015-SF-120).

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Authors and Affiliations

  1. Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining, People’s Republic of China

    Yuwei Wang, Yourui Suo, Yanan Sun & Jinmao You

  2. State Key Laboratory of Plateau Ecology and Agriculture, Qinghai University, Xining, People’s Republic of China

    Yourui Suo & Jinmao You

  3. University of the Chinese Academy of Sciences, Beijing, People’s Republic of China

    Yuwei Wang & Yanan Sun

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  1. Yuwei Wang
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  2. Yourui Suo
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Correspondence to Yourui Suo.

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Wang, Y., Suo, Y., Sun, Y. et al. Determination of Triterpene Acids from 37 Different Varieties of Raspberry using Pre-column Derivatization and HPLC Fluorescence Detection. Chromatographia 79, 1515–1525 (2016). https://doi.org/10.1007/s10337-016-3174-1

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  • DOI: https://doi.org/10.1007/s10337-016-3174-1

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