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Food Analytical Methods

, Volume 10, Issue 10, pp 3225–3234 | Cite as

Phytosterol Determination and Method Validation for Selected Nuts and Seeds

  • Md. Atiqual Islam
  • Beom-Gyun Jeong
  • Jiyoung Jung
  • Eui-Cheol Shin
  • Sung-Gil Choi
  • Jiyeon ChunEmail author
Article

Abstract

A method involving alkali and/or acid hydrolysis of phytosterols followed by trimethylsilyl ether derivatization coupled with GC-FID analysis was validated and applied in the analysis of major phytosterols (campesterol, stigmasterol, β-sitosterol, and Δ5-avenasterol) in nuts (n = 7), seeds (n = 9), legumes (n = 2), and grain (n = 1). The acid-labile Δ5-avenasterol was extracted with alkaline hydrolysis only before derivatization. Quantification of all phytosterols was done using the computed relative response factor of 5α-cholestane (internal standard). Analyses of internal and external phytosterol standards showed good linearity for all phytosterols (R 2 of 0.999); LOD and LOQ of phytosterols were determined to be 0.01–0.12 and 0.04–0.40 mg/100 g, respectively. Repeatability and reproducibility precision analyses showed acceptable coefficient of variation of less than 3 and 4%, respectively, and satisfactory Horwitz ratio values of <1.0. Excellent accuracy was proved by the high recovery values of 91.4–106.0% for campesterol, β-sitosterol, and stigmasterol. Δ5-Avenasterol, the most oxidation-susceptible sterol, showed a recovery of about 60%. The total phytosterol (sum of major phytosterols quantified) contents in the 19 samples varied from 38.8 mg/100 g (white quinoa seed) to 246.2 mg/100 g (sunflower seed). β-Sitosterol was the predominant phytosterol (54–86.0% of total) among all samples except fennel seed in which stigmasterol was predominant. Analytical quality control chart maintained during the study period showed that assays were performed under control. Method validation indicated that the analytical method can be applied for accurate determination of campesterol, β-sitosterol, and stigmasterol in selected food samples.

Keywords

Phytosterols Nuts Method validation Acid hydrolysis Saponification Double hydrolysis 

Notes

Compliance with Ethical Standards

Conflict of Interest

Md. Atiqual Islam declares that he has no conflict of interest. Beom-Gyun Jeong declares that he has no conflict of interest. Jiyoung Jung declares that she has no conflict of interest. Eui-Cheol Shin declares that he has no conflict of interest. Sung-Gil Choi declares that he has no conflict of interest. Jiyeon Chun declares that she has no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants performed by the authors.

Informed Consent

Not applicable.

References

  1. Andersson SW, Skinner J, Ellegard L, Welch AA, Bingham S, Mulligan A, Andersson H, Shaw K (2004) Intake of dietary plant sterols is inversely related to serum cholesterol concentration in men and women in the EPIC Norfolk population: a cross-sectional study. Eur J Clin Nutr 58:1378–1385CrossRefGoogle Scholar
  2. Association of Official Analytical Chemists (2002) AOAC Guidelines for Single Laboratory Validation of Chemical Analytical MethodGoogle Scholar
  3. Awad AB, Fink CS, Williams H, Kim U (2001) In vitro and in vivo (SCID mice) effects of phytosterols on the growth and dissemination of human prostate cancer PC-3 cells. Eur J Cancer Prev 10:507–513CrossRefGoogle Scholar
  4. Awad AB, Chan KC, Downie AC, Fink CS (2000a) Peanuts as a source of beta-sitosterol, a sterol with anticancer properties. Nutr Cancer 36:238–241CrossRefGoogle Scholar
  5. Awad AB, Downie A, Fink CS, Kim U (2000b) Dietary phytosterol inhibits the growth and metastasis of MDA-MB-231 human breast cancer cells grown in SCID mice. Anticancer Res 20:821–824Google Scholar
  6. Boskou D, Morton ID (1975) Changes in the sterol composition of olive oil on heating. J Sci Fd Agric 26:1149–1153CrossRefGoogle Scholar
  7. Brufau G, Canela MA, Rafecas M (2008) Phytosterols: physiologic and metabolic aspects related to cholesterol-lowering properties. Nutr Res 28:217–225CrossRefGoogle Scholar
  8. Campos-Mondragon MG, Calderon De La Barca AM, Dur-Prado A, Campos-Reyes LC, Oliart-Ros RM, Orteg-Garcıa J, Medina-Juarez LA, Angulo O (2009) Nutritional composition of new peanut (Arachis hypogaeaL.) cultivars. Grasas Y. Aceites 60:161–167CrossRefGoogle Scholar
  9. CFR 21CFR101.83 (2016) Code of Federal Regulations Title 21, Department of Health and Human Services, United Stated Food and Drug AdministrationGoogle Scholar
  10. Christiansen LI, Lahteenmaki PL, Mannelin MR, Seppanen-Laakso TE, Hiltunen RV, Yliruusi JK (2001) Cholesterol-lowering effect of spreads enriched with microcrystalline plant sterols in hypercholesterolemic subjects. Eur J Nutr 40:66–73CrossRefGoogle Scholar
  11. Delgado-Zamarreno MM, Fernandez-Prieto C, Bustamante-Rangel M, Perez-Martin L (2016) Determination of tocopherols and sitosterols in seeds and nuts by QuEChERS-liquid chromatography. Food Chem 192:825–830CrossRefGoogle Scholar
  12. Duchateau GSMJE, Janssen HGM, Louter AJH (2004) Phytosterols as Functional Food Components and Nutraceuticals. In: Dutta PC (ed) Plant sterol analysis in relation to functional foods. Marcel Dekker, Inc, New York, pp 75–131Google Scholar
  13. Ellegǻrd L, Bosaeus I, Andersson H (2000) Will recommended changes in fat and fibre intake affect cholesterol absorption and sterol excretion? An ileostomy study. Eur J Clin Nutr 54:306–313CrossRefGoogle Scholar
  14. European Food Safety Authority (EFSA) (2008) Plant sterols and blood cholesterol—scientific substantiation of a health claim related to plant sterols and lower/reduced blood cholesterol and reduced risk of (coronary) heart disease pursuant to Article 14 of Regulation (EC) No 1924/2006. EFSA J 6(8):781–n/aCrossRefGoogle Scholar
  15. Fenton M (1992) Chromatographic separation of cholesterol in foods. J Chromatogr A 624:369–388CrossRefGoogle Scholar
  16. Fernández-Cuesta A, Fernández-Martínez JM, Velasco L (2013) Variation for seed phytosterols in a set of safflower cultivars. J Food Agric Environ 11:656–660Google Scholar
  17. Grosso NR, Guzman CA (1995) Chemical composition of aboriginal peanut (Arachis hypogaea L.) seeds from Peru. J Agric Food Chem 43:102–105CrossRefGoogle Scholar
  18. Horwitz W, Albert R (2006) The Horwitz ratio (HorRat): a useful index of method performance with respect to precision. J AOAC Int 89:1095–1109Google Scholar
  19. Jekel AA, Vaessen HAMG, Schothorst RC (1998) Capillary gas-chromatographic method for determining non-derivatized sterols—some results for duplicate 24 h diet samples collected in 1994. Fresen J Anal Chem 360:595–600CrossRefGoogle Scholar
  20. Jonnala RS, Dunford NT, Chenault K (2006a) Tocopherol, phytosterol and phospholipid compositions of genetically modified peanut varieties. J Sci Food Agr 86:473–476CrossRefGoogle Scholar
  21. Jonnala RS, Dunford NT, Dashiell KE (2006b) Tocopherol, phytosterol and phospholipid compositions of new high oleic peanut cultivars. J Food Comp Anal 19:601–605CrossRefGoogle Scholar
  22. Kamal-Eldin A, Maatta K, Toivo J, Lampi AM, Piironen V (1998) Acid-catalyzed isomerization of fucosterol and delta5-avenasterol. Lipids 33:1073–1077CrossRefGoogle Scholar
  23. Kendall CWC, Jenkins DJA (2004) A dietary portfolio: maximal reduction of low-density lipoprotein cholesterol with diet. Curr Atheroscler Rep 6:492–498CrossRefGoogle Scholar
  24. Kesselmeier J, Eichenberger W, Urban B (1985) High performance liquid chromatography of molecular species from free sterols and sterylglycosides isolated from oat leaves and seeds. Plant Cell Physiol 26:463–471CrossRefGoogle Scholar
  25. KS1320 (2013) Walnuts, almonds and pistachios. GAIN Report– Republic of Korea. USDA Foreign Agricultural ServiceGoogle Scholar
  26. KS8042 (2008) Almonds, walnuts and pistachios. GAIN Report– Republic of Korea. USDA Foreign Agricultural ServiceGoogle Scholar
  27. Laakso P (2005) Analysis of sterols from various food matrices. Eur J Lipid Sci Tech 107:402–410CrossRefGoogle Scholar
  28. Lagarda MJ, García-Llatas G, Farré R (2006) Analysis of phytosterols in foods. J Pharm Biomed Anal 41:1486–1496CrossRefGoogle Scholar
  29. Lampi AM, Piironen V (2004) Analysis of phytosterols in foods. In: Dutta PC (ed) Phytosterols as functional food components and Nutraceuticals. Marcel Dekker, Inc., New York, pp 33–73Google Scholar
  30. Loizou S, Lekakis I, Chrousos GP, Moutsatsou P (2010) Beta-sitosterol exhibits anti-inflammatory activity in human aortic endothelial cells. Mol Nutr Food Res 54:551–558CrossRefGoogle Scholar
  31. MacKay DS, Jones PJH (2011) Phytosterols in human nutrition: type, formulation, delivery, and physiological function. Eur J Lipid Sci Tech 113:1427–1432CrossRefGoogle Scholar
  32. Moreau RA, Whitaker BD, Hicks KB (2002) Phytosterols, phytostanols, and their conjugates in foods: structural diversity, quantitative analysis, and health-promoting uses. Prog Lipid Res 41:457–500CrossRefGoogle Scholar
  33. Ostlund RE Jr (2002) Phytosterols in human nutrition. Annu Rev Nutr 22:533–549CrossRefGoogle Scholar
  34. Phillips KM, Ruggio DM, Ashraf-Khorassani M (2005a) Analysis of steryl glucosides in foods dietary supplements by solid-phase extraction and gas chromatography. J Food Lipids 12:124–140CrossRefGoogle Scholar
  35. Phillips KM, Ruggio DM, Ashraf-Khorassani M (2005b) Phytosterol composition of nuts and seeds commonly consumed in the United States. J Agric Food Chem 53:9436–9445CrossRefGoogle Scholar
  36. Piironen V, Toivo J, Lampi A (2000) Natural sources of dietary plant sterols. J Food Comp Anal 13:619–624CrossRefGoogle Scholar
  37. Shin EC, Pegg RB, Phillips RD, Eitenmiller RR (2010) Commercial peanut (Arachis hypogaea L.) cultivars in the United States: phytosterol composition. J Agric Food Chem 58:9137–9146CrossRefGoogle Scholar
  38. Sorenson WR, Sullivan D (2007) Determination of campesterol, stigmasterol, and beta-sitosterol in saw palmetto raw materials and dietary supplements by gas chromatography: collaborative study. J AOAC Int 90:670–678Google Scholar
  39. Srigley CT, Haile EA (2015) Quantification of plant sterols/stanols in foods and dietary supplements containing added phytosterols. J Food Comp Anal 40:163–176CrossRefGoogle Scholar
  40. Toivo J, Phillips K, Lampi A, Piironen V (2001) Determination of sterols in foods: recovery of free, esterified, and glycosidic sterols. J Food Comp Anal 14:631–643CrossRefGoogle Scholar
  41. Toivo J, Lampi A, Aalto S, Piironen V (2000) Factors affecting sample preparation in the gas chromatographic determination of plant sterols in whole wheat flour. Food Chem 68:239–245CrossRefGoogle Scholar
  42. USDA SR28 (2015) National Nutrient Database for Standard Reference, Release 28. In: US Department of Agriculture, Agricultural Research Service, Nutrient Data Laboratory. http://www.ars.usda.gov/ba/bhnrc/ndl

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Md. Atiqual Islam
    • 1
  • Beom-Gyun Jeong
    • 1
  • Jiyoung Jung
    • 1
  • Eui-Cheol Shin
    • 2
  • Sung-Gil Choi
    • 3
  • Jiyeon Chun
    • 1
    Email author
  1. 1.Department of Food Science and TechnologySunchon National UniversitySuncheonSouth Korea
  2. 2.Department of Food ScienceGyeongnam National University of Science and TechnologyJinjuSouth Korea
  3. 3.Department of Food Science and TechnologyGyeongsang National UniversityJinjuSouth Korea

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