Download PDF
Planta Med 2009; 75(7): 704-710
DOI: 10.1055/s-0029-1185439
Plant Analysis
Mini-Review
© Georg Thieme Verlag KG Stuttgart · New York

Influence of Sample Preparation on the Assay of Isoflavones

Devanand L. Luthria1 , Savithiry S. Natarajan2
  • 1Food Composition and Methods Development Laboratory, Beltsville Human Nutrition Research Center, ARS, USDA, Beltsville, Maryland, USA
  • 2Soybean Genomics and Improvement Laboratory, Plant Sciences Institute, ARS, USDA, Beltsville, Maryland, USA
Further Information

Publication History

received October 7, 2008 revised January 12, 2009

accepted January 21, 2009

Publication Date:
13 March 2009 (online)

   Permissions and Reprints

Abstract

The complexity of sample matrices, coexistence of multiple forms of bioactive phytochemicals, and their interaction of with other cellular components pose a significant challenge for optimized extraction and accurate estimation of bioactive phytochemicals in foods and dietary supplements. This article discusses the significance of optimizing extraction procedures for accurate assay of phytochemicals from different matrices using bioactive isoflavones as model substrate because isoflavones are known to exist in nature as free aglycones or as conjugates with sugars and/or acids. The wide structural diversity and polarities of free and conjugated isoflavones makes optimum extraction and accurate quantification of isoflavones a challenging task. This paper reviews variables, extraction solvent composition (aqueous-organic solvents mixtures at different acidification levels), physical extraction techniques (Soxhlet, stirring, ultrasonic, microwave, pressurized, supercritical-fluid, high-speed counter-current chromatography), and parameters (temperature, pressure, number of cycles, solid-solvent ratio) that influence quantitative extraction of isoflavones from different matrices. In addition, this review covers a brief overview of structures, sources, bioactivities, separation, and detection used for isoflavones analysis. Optimum extraction efficiencies of isoflavones were obtained with EtOH : H2O : DMSO (70 : 25 : 5, v/v/v) as the extraction solvent and acidification of extraction solvent favored partial degradation of conjugated forms to their corresponding aglycones. Accurate quantification of isoflavones in foods, plants, and dietary supplements will allow researchers and regulators to provide more precise guidelines on dietary intake and safety levels necessary to achieve optimum health.

Key words

isoflavones - extraction solvents composition - extraction techniques and conditions - sample preparation - foods and plants matrices

References

  • 1 Habauzit V, Horcajada M-N. Phenolic phytochemicals and bone.  Phytochem Rev. 2008;  7 313-344
    CrossrefPubMedGoogle Scholar
  • 2 Chen L, Vigneault C, Vijaya Raghavan G S, Kubow S. Importance of the phytochemical contents of fruits and vegetables to human health.  Stewart Postharvest Rev. 2007;  3 1-5
    PubMedGoogle Scholar
  • 3 Hasler C M. Functional foods: their role in disease prevention and health promotion.  Food Technol. 1998;  52 63-70
    PubMedGoogle Scholar
  • 4 Mark-Herbert C. Innovation of a new product category-functional foods.  Technovation. 2004;  24 713-719
    CrossrefPubMedGoogle Scholar
  • 5 Wansink B, Westgren R E, Cheney M W. Hierarchy of nutritional knowledge that relates to consumption of functional foods.  Nutrition. 2005;  21 264-268
    CrossrefPubMedGoogle Scholar
  • 6 Wildman R EC. Nutraceuticals: a brief review of historical and teleological aspects. Wildman REC Handbook of nutraceuticals and functional foods. New York; CRC Press 2001: 1-12
    Google Scholar
  • 7 Luthria D L. Significance of sample preparation in developing analytical methodologies for accurate estimation of bioactive compounds in functional foods.  J Sci Food Agric. 2006;  86 2266-2272
    CrossrefPubMedGoogle Scholar
  • 8 Majors R E. An overview of sample preparation methods for solids.  LC‐GC. 1999;  17 S8-S13
    PubMedGoogle Scholar
  • 9 Escarpa A, Gonzalez M C. An overview of analytical chemistry of phenolic compounds in foods.  Critical Rev Anal Chem. 2001;  31 57-139
    PubMedGoogle Scholar
  • 10 Kaczk M, Shahidi F. Extraction and analysis of phenolics in food.  J Chromatogr A. 2004;  1054 95-111
    PubMedGoogle Scholar
  • 11 Rijke E, Out P, Niessen W MA, Ariese F, Gooijer C, Brinkman U AT. Analytical separation and detection methods for flavonoids.  J Chromatogr A. 2006;  1112 31-63
    CrossrefPubMedGoogle Scholar
  • 12 Stalikas C D. Extraction, separation, and detection methods for phenolic acids and flavonoids.  J Sep Sci. 2007;  30 3268-3295
    CrossrefPubMedGoogle Scholar
  • 13 Toebes A HW, Boer V D, Verkleij J AC, Lingeman H, Ernst W HO. Extraction of isoflavone malonylglucosides from Trifolium pretense L. .  J Agric Food Chem. 2005;  53 4660-4666
    CrossrefPubMedGoogle Scholar
  • 14 Penalvo J L, Nurmi T, Adlercreutz H. A simplified HPLC method for total isoflavones in soy products.  Food Chem. 2004;  87 297-305
    CrossrefPubMedGoogle Scholar
  • 15 Ralston L, Subramanian S, Matsuno M, Yu O. Partial reconstruction of flavonoid and isoflavonoid biosynthesis in yeast using soybean type I and type II chalcone isomerases.  Plant Physiol. 2005;  137 1375-1388
    CrossrefPubMedGoogle Scholar
  • 16 Elbridge A C, Kwolek W F. Soybean isoflavones: effect of environment and variety on composition.  J Agric Food Chem. 1983;  31 394-396
    CrossrefPubMedGoogle Scholar
  • 17 Liggins J, Bluck L JC, Runswick S, Atkinson C, Coward W A, Bingham S A. Daidzein and genistein content of fruits and nuts.  J Nutr Biochem. 2000;  11 326-331
    PubMedGoogle Scholar
  • 18 Liggins J, Bluck L JC, Runswick S, Atkinson C, Coward W A, Bingham S A. Daidzein and genistein content of vegetables.  Br J Nutr. 2000;  84 717-725
    PubMedGoogle Scholar
  • 19 Liggins J, Mulligan A, Runswick S, Bingham S A. Daidzein and genistein content of cereals.  Eur J Clin Nutr. 2002;  56 961-966
    CrossrefPubMedGoogle Scholar
  • 20 Tsao R, Yang R, Young J C. Antioxidant isoflavones in osage orange, Maclura pomifera (Raf.) Schneid.  J Agric Food Chem. 2003;  51 6445-6451
    CrossrefPubMedGoogle Scholar
  • 21 Wu Q, Wang M, Simon J E. Analytical methods to determine phytoestrogenic compounds.  J Chromatogr B. 2004;  812 325-355
    PubMedGoogle Scholar
  • 22 Adlercreutz H, Heinonen S M, Penalvo-Garcia J. Phytoestrogens, cancer and coronary heart disease.  Biofactors. 2004;  22 229-236
    CrossrefPubMedGoogle Scholar
  • 23 Caragay A B. Cancer preventive foods and ingredients.  Food Technol. 1992;  46 65-68
    PubMedGoogle Scholar
  • 24 Messina M. Soy phytoestrogens (isoflavones) and breast cancer.  Am J Clin Nutr. 1999;  70 574-575
    PubMedGoogle Scholar
  • 25 Anderson J JB, Garner S C. Phytoestrogens and human function.  Nutr Today. 1997;  32 232-239
    PubMedGoogle Scholar
  • 26 Park D, Huang T, Frishman W H. Phytoestrogens as cardioprotective agents.  Cardiol Rev. 2005;  13 13-17
    PubMedGoogle Scholar
  • 27 Valachovicova T, Slivova V, Sliva D. Cellular and physiological effects of soy isoflavonoids.  Mini Rev Med Chem. 2004;  4 881-887
    PubMedGoogle Scholar
  • 28 Branca F, Lorenzetti S. Health effects of phytoestrogens.  Forum Nutr. 2005;  57 100-111
    PubMedGoogle Scholar
  • 29 Lin F, Giusti M M. Effects of solvent polarity and acidity on the extraction efficiency of isoflavones from soybeans (Glycine max).  J Agric Food Chem. 2005;  53 3795-3800
    CrossrefPubMedGoogle Scholar
  • 30 He J, Zhao Z, Shi Z, Zhao M, Li Y, Chang W. Analysis of isoflavone daidzein in Puerariae Radix with micelle-mediated extraction and preconcentration.  J Agric Food Chem. 2005;  53 518-523
    CrossrefPubMedGoogle Scholar
  • 31 Lee M H, Lin C C. Comparison of techniques for extraction of isoflavones from the roots of Radix Puerariae: ultrasonic and pressurized solvent extractions.  Food Chem. 2007;  105 223-228
    CrossrefPubMedGoogle Scholar
  • 32 Luthria D, Vinjamoori D, Noel K. Effect of moisture content, grinding, and extraction technologies on assay of crude fat. Luthria DL Oil extraction and analysis: critical issues and comparative studies. Illinois; AOCS Press 2004: 25-39
    Google Scholar
  • 33 Mukhopadhyay S, Luthria D L, Robbins R J. Optimization of extraction process for phenolic acids from black cohosh (Cumicifuga racemosa) by pressurized liquid extraction.  J Sci Food Agric. 2006;  86 156-162
    CrossrefPubMedGoogle Scholar
  • 34 Luthria D L, Biswas R, Natarajan S S. Comparison of extraction solvents and techniques used for the assay of isoflavones from soybeans.  Food Chem. 2007;  105 325-333
    CrossrefPubMedGoogle Scholar
  • 35 Rostagno M A, Palma M, Barroso C G. Ultrasound-assisted extraction of isoflavones from soy beverages blended with fruit juices.  Anal Chim Acta. 2007;  597 265-272
    CrossrefPubMedGoogle Scholar
  • 36 Rostagno M A, Palma M, Barroso C G. Microwave assisted extraction of soy isoflavones.  Anal Chim Acta. 2007;  588 274-282
    CrossrefPubMedGoogle Scholar
  • 37 Rostagno M A, Palma M, Barroso C G. Solid-phase extraction of isoflavones.  J Chromatogr A. 2005;  1076 110-117
    CrossrefPubMedGoogle Scholar
  • 38 Du Q, Li Z, Ito Y. Preparative separation of isoflavone components in soybeans using high-speed counter-current chromatography.  J Chromatogr A. 2001;  923 271-274
    CrossrefPubMedGoogle Scholar
  • 39 Cao X, Tian Y, Zhang T, Li X, Ito Y. Separation and purification of isoflavones from Pueraria lobata by high speed counter-current chromatography.  J Chromatogr A. 1999;  855 709-713
    CrossrefPubMedGoogle Scholar
  • 40 Ye H, Chen L, Li Y, Peng A, Fu A, Song H, Tang M, Luo H, Luo Y, Xu Y, Shi J, Wei Y. Preparative isolation and purification of three rotenoids and one isoflavones from the seeds of Millettia pachycarpa Benth by high-speed counter-current chromatography.  J Chromatogr A. 2008;  1178 101-107
    CrossrefPubMedGoogle Scholar
  • 41 Xu H N, He C H. Extraction of isoflavones from stem of Pueraria lobata (Willd.) Ohwi using n-butanol/water two-phase solvent system and separation of daidzein.  Sep Purif Technol. 2007;  56 85-89
    CrossrefPubMedGoogle Scholar
  • 42 Kao T H, Chien J T, Chen B H. Extraction yield of isoflavones from soybean cake as affected by solvent and supercritical carbon dioxide.  Food Chem. 2008;  107 1728-1736
    CrossrefPubMedGoogle Scholar
  • 43 Zuo Y B, Zeng A W, Yuan X G, Yu K T. Extraction of soybean isoflavones from soybean meal with aqueous methanol modified supercritical carbon dioxide.  J Food Eng. 2008;  89 384-389
    CrossrefPubMedGoogle Scholar
  • 44 Ksycinska H, Sobik B, Popiolkiewicz J, Polkowski K, Krzeczynski P, Ramza J, Pucko W, Grynkiewicz G. Determination of new derivative of genistein in culture media by liquid chromatography.  J Chromatogr B. 2004;  799 217-231
    CrossrefPubMedGoogle Scholar
  • 45 Grace P B, Mistry N S, Carter M H, Leathem A J, Teale P. High throughput quantification of phytoestrogens in human urine and serum using liquid chromatography/tandem mass spectrometry (LC‐MS/MS).  J Chromatogr B. 2007;  853 138-146
    CrossrefPubMedGoogle Scholar
  • 46 Otieno D O, Rose H, Shah N P. Profiling and quantification of isoflavones in soy milk from soy protein isolate using extracted ion chromatography and positive ion fragmentation techniques.  Food Chem. 2007;  105 1642-1651
    CrossrefPubMedGoogle Scholar
  • 47 Maul R, Schebb N H, Kulling S E. Application of LC and GC hyphenated with mass spectrometry as tool for characterization of unknown derivatives of isoflavonoids.  Anal Bioanal Chem. 2008;  DOI: 10.1007/s00216-008-1884-2008 4 1-12
    PubMedGoogle Scholar
  • 48 Wilkinson A P, Wähälä K, Williamson G. Identification and quantification of polyphenol phytoestrogens in foods and human biological fluids.  J Chromatogr B. 2002;  759 33-41
    PubMedGoogle Scholar
  • 49 Grynkiewicz G, Ksycinska H, Ramza J, Zagrodzka J. Chromatographic quantification of isoflavones.  Acta Chromatogr. 2005;  15 31-65
    PubMedGoogle Scholar
  • 50 Vacek J, Klejdus B, Lojkova L, Kuban V. Current trends in isolation, separation, determenination and identification of isoflavones: a review.  J Sep Sci. 2008;  31 2054-2067
    CrossrefPubMedGoogle Scholar
  • 51 Zhai X J, Qu H B, Shao Q, Cheng Y Y. Simultaneous quantification of puerarin and daidzein in rat plasma by high-performance liquid chromatography with post-column modification and fluorescence detection.  Chromatgraphia. 2007;  66 43-47
    PubMedGoogle Scholar
  • 52 Careri M, Corradini C, Elviri L, Mangia A. Optimization of a rapid microwave assisted ectraction method for liquid chromatography-electrospray-tandem mass spectrometry determination of isoflavonoid aglycones in soybeans.  J Chromatogr A. 2007;  1152 274-279
    CrossrefPubMedGoogle Scholar
  • 53 Lv Y Q, Tan T W, Wang M Y, Janson J C. One step rapid determination and purification of puerarin from Radix Puerariae using n-octylamine modified poly (methacrylate-co-dimethacrylate) monolith.  J Chromatogr B. 2008;  871 1-6
    CrossrefPubMedGoogle Scholar
  • 54 Murphy P A, Barua K, Hauck C C. Solvent extraction selection in the determination of isoflavones in soy foods.  J Chromatogr B. 2002;  777 129-138
    CrossrefPubMedGoogle Scholar
  • 55 Rostagno M A, Araujo J MA, Sandi D. Supercritical fluid extraction of isoflavones from soybean flour.  Food Chem. 2002;  78 111-117
    CrossrefPubMedGoogle Scholar
  • 56 Lee J H, Renita M, Fioritto R J, Martin S K, Schwartz S J, Vodovotz Y. Isoflavone characterization and antioxidant activity of Ohio soybeans.  J Agric Food Chem. 2004;  52 2647-2651
    CrossrefPubMedGoogle Scholar
  • 57 Li-Hsun C, Ya-Chuan C, Chieh-Ming C. Extracting and purifying isoflavones from defatted soybean flakes using superheated water at elevated pressures.  Food Chem. 2004;  84 279-285
    CrossrefPubMedGoogle Scholar
  • 58 Klejdus B, Mikelova R, Adam V, Zehnalek J, Vacek J, Kizek R, Kuban V. Liquid chromatographic-mass spectrophotometric determination of genistin and daidzin in soybean food samples after accelerated solvent extraction with modified content of extraction cell.  Anal Chim Acta. 2004;  517 1-11
    CrossrefPubMedGoogle Scholar
  • 59 Klejdus B, Mikelova R, Petrlova J, Potesil D, Adam V, Stiborova M, Hodek P, Vacek J, Kizek R, Kuban V. Evaluation of isoflavone aglycon and glycoside distribution in soy plants and soybeans by fast column high-performance liquid chromatography coupled with a diode-array detector.  J Agric Food Chem. 2005;  53 5848-5852
    CrossrefPubMedGoogle Scholar
  • 60 Achouri A, Boye J I, Belanger D. Soybean isoflavones: efficacy of extraction conditions and effect of food type on extractability.  Food Res Int. 2005;  38 1199-1204
    CrossrefPubMedGoogle Scholar

Devanand Luthria

Food Composition and Methods Development Laboratory
Room 202 B, Building 161, BARC‐E
Beltsville Human Nutrition Research Center
USDA

10300 Baltimore Ave.

Beltsville

MD 20705-3000

USA

Phone: + 1 30 15 04 72 47

Fax: + 1 30 15 04 83 14

Email: D.Luthria@ars.usda.gov

      >