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


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.


Phytosterols Nuts Method validation Acid hydrolysis Saponification Double hydrolysis 


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.


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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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