Measurement and Quality Control of Fatty Acid and trans Fats Determination in Soybeans and Soybean Oil

1. Measurement and Quality Control of Fatty Acid and trans Fats Determination in Soybeans and Soybean Oil Gina M. Clapper AOCS Technical Services Low Linolenic Acid Soybeans and trans Fatty Acids AOAC International Midwest Section Annual Meeting and Exposition Kansas City, Missouri, USA

2. January 1, 2006 all food labels must contain the trans Fatty Acid content if level is at or above 0.5g graphic from FDA website

3. Options for the food industry? • Keep existing formulations and label appropriately • Find alternatives to partially hydrogenated oils • PH-Soybean oil is most common

4. The Importance of the SQT Initiative Raising the Bar on Beans

5. What is SQT? • A USB/AOCS collaboration • Supports the mission of the checkoff-funded Better Bean Initiative (BBI) to accelerate the development and availability of soybean seed with enhanced compositional traits

6. Why SQT? • Observations • Commercial testing labs were using different methods than breeder and seed company laboratories. • NOPA Certified Laboratories use AOCS Methods and analyze > 70% of the referee SBM contracted in the US and use AOCS methods for oilseed analyses • Oilseed Processors predominantly use AOCS methods • Equipment Manufacturers, Seed breeders and Seed Companies generally use internal methods • Inability to identify, verify, and quantify enhanced soybean traits limits industry growth in this area • Unrealized value and profit from enhanced soybean strains

7. Why SQT? (continued) • SQT creates and facilitates • Systems of verification • Reliable analytical results • Opportunity for all stakeholders in the soy industry to agree

8. SQT Mission • To guarantee quality analytical results at all levels in the soybean industry • To increase confidence in the ability to identify soybeans with enhanced quality traits • To achieve, maintain, and promote peak levels of laboratory accuracy through industry participation in a proficiency program

9. Equipment Manufacturers S Q T S Q T S Q T S Q T Processors Fats and Oil Professionals Elevators & Grain Handlers Seed Breeders Farmers End User Laboratories Referee/Contract Laboratories Seed Companies

10. Key Concept: Verification • Systematic verification of analytical measurements • Different sectors create potential for different methodologies • Primary methods will be implemented throughout soy continuum • First time industry will be standardized

11. How do we achieve this? • Analyze samples regularly • Establish industry-wide acceptance of analytical methods and protocols and their implementation under internationally accepted quality management standards • Determine laboratory performance in terms of accuracy and bias

12. Key Concept: Certification • Certify value-added enhanced soybean varieties for the food and feed industries • Identify specific physical traits • Validate analytically the presence of these traits • Certify the identity of enhanced varieties throughout the continuum

13. Advantages of SQT • Defines standards for quality assurance • Provides quality testing methods • Validates industry infrastructure • Sets expectations for analytical results • Provides traceable standards, reference material, consensus standards

14. Advantages of SQT (continued) • Promotes and markets testing standards within the industry • Increases consumer confidence with system of checks and balances • Decreases risk to buyers industry-wide • Realizes added value of American enhanced soybeans

15. Benefits of SQT • Increase the competitiveness of American soybeans in the global marketplace • Generate a high level of confidence in enhanced soybean products • Better position U.S. soybeans to meet the needs of oil and protein end users and improve the economic value for the U.S. soybean industry

16. SQT Phases 1 Primary Methods Secondary Methods Identify Participants & Needs 3 2 SQT Proficiency Program 4 Laboratory Quality Assurance 5 ISO 17025 Certification 6

17. Phase One • Identify analysis methods for • Protein content • Oil content • Fatty acid composition analysis (completed)

18. Phase Two • Develop and validate methods of analysis including the evaluation of secondary methods • Library of reference samples to be used for calibrating non-destructive technologies (NIR currently)

19. New methods on the horizon • Amino Acids • Phytate • Isoflavones

20. Phase Three • Identify users and their requirements • Seed companies • Referee and private laboratories • End-user laboratories • Elevator and crop handling facilities • Establish a core group of expert laboratories

21. Phase Four • Develop Soybean Quality Traits Laboratory Program, including use of proficiency testing and standards • 4 samples • 6 times/year • Initiated November 2003

22. Standards, Methods, and Proficiency Testing • Members-only secure access to required methodologies • Secure members-only access to • Online protocols • Secure electronic data reporting forms (can enter moisture, protein, oil and FA profile, report tables) • Web- and email-based status, results, proficiency, and rankings reports

23. Phase Five • Implement laboratory quality assurance • Standard methods • Certification • Proficiency participation • Results monitoring

24. Phase Six • Incorporate SQT methods of analysis into ISO 17025 certification and quality audits

25. For those who still need to label…

26. Evaluation of current testing methods for Linolenic Acid content in soybeans • GIPSA (KCMO) evaluated AOCS Official Method Ce 1-62 to determine it was appropriate for determining low-lin oil • Internal and commercial laboratories participated in a collaborative study • Preliminary results indicate that Ce 1-62 is reasonably accurate, precise and rugged

27. Collaborative Study Report Analysis of Fatty Acid Composition PROJECT: Ce-1h-05 The Analysis of Fatty Acid Methyl Esters

28. New Method Study: AOCS Official Method Ce 1h-05 Standardization of the current method(AOCS Ce 1f-96): • Internal Standard (C21:0) • Column type (SP-2560 or CP-Sil 88) • Column length (100m) • Carrier gas (Hydrogen or Helium) following slides borrowed from Dawn McCluckie

29. PARTICIPATION - Samples to 35 labs worldwide - 21 responded, 11 were used TEST MATERIALS - 5 reference samples with labeled chromatograms • 10 randomly coded samples REPORTING - Run in duplicate - Results  Area, Area%, TG, and FA - Labeled chromatograms

30. Examples of why some participants were eliminated: • 1 lab  wrong column • 1 lab  wrong carrier gas • 1 lab  wrong column length • 1 lab  only Area % values

31. ODD OUTLIERS Non-reproducible data i.e. not enough responses for statistical analysis, so all reported values considered outliers REGULAR OUTLIERS Analyzable data Cochran Single Grubbs Double Grubbs Results- Outliers

32. Odd Outlier Example • Sample 2 – margarine oil • C21:0 internal standard • C22:0 • C23:0 peak present • C24:0

34. Conclusions from this study • The peaks are present on the chromatogram  The method is good and allows for peak separation  Training may be necessary for proper peak identification

35. AOCS Official Method Ce 1h-05 • Approved by Uniform Methods Committee at the AOCS Annual Meeting • Will be available in June with: • Labeled chromatograms and reference samples • Interactive Excel spreadsheet for calculations • Study samples will be released as consensus reference material

36. Training • AOCS will offer hands-on training workshop for all participants in the collaborative study and methodology meeting • AOCS will offer training workshops for the public late summer and into fall to assist industry’s efforts to comply with FDA labeling regulations

37. Thank you for your attention! Contact AOCS Technical Services at: technical@aocs.org www.aocs.org/tech