3.  Company Philosophy - Whatever you do, work at it with all your heart, as working for the Lord, not for men… (Colossians 3:23)  Goal is to develop products that improve existing analytical methods and reduce costs.  “Simplicity of design is the result of profound thought” (William Hazzlet)  Two major product lines: Fiber and Fat analysis  High emphasis on analytical and technical support – Total Satisfaction

4. International Distribution  Over 30 Formal Distributors  Over 30 Informal Distributors –Long Term Relationship – Since 2000 –Shares Customer Focus –Invested in Product Knowledge & Expertise

6.  ANKOM TDF Dietary Fiber  Same Chemistry  Consistent Automated Process  Reduces Technician Variability  Enhanced Mixing  Continuous throughout digestion  Reduced Filtration Time  IDF = 30 - 90 seconds  SDF = 1 - 5 minutes Overview

7. Transfer to Filter - rinse and scrape beaker as needed 14 Transfer to Filter - rinse and scrape beaker as needed 21 Prepare for Test Prepare Crucibles1 Weigh Samples3Sample to 400 mL beaker4Put Sample into Bath5 Add 2 ml Amylase at 95° Monitor 30 minutes Add 10 ml H 2 O Add Protease at 60° Monitor 30 minutes Add 5 ml.561N HCl at 60° Add 1-5 ml AMG at 60° Monitor 30 minutes 6 7 8 9 10 11 12 13 Add 40 ml Buffer5 Digest Sample Add 250 ml 95% ETOH at 60° Wait 60 minutes 19 20 Solute Add 2x10 ml H 2 O at 70°15 Add 2x15 ml 78% ETOH16 Add 2x15 ml 95%ETOH 17 Add 2x15 ml Acetone18 IDF Extract IDF Add 2x15 ml 78% ETOH22 Add 2x15 ml 95% ETOH23 Add 2x15 ml Acetone24 SDF Extract SDF Ash Overnight1a Clean with Vacuum 1bSoak 1hr @ Room Temp1c Rinse with 3 solutions1d Add DE, Dry @ 130°C1eCool, 1 hour, weigh1fPrepare Water bath/95°C2 AOAC 991.43 - AACC 32.07.01 Conventional Protein Correction25Ash Correction26 Add all actions including IDF and SDF drying/weighing Total = 35 steps

8. AOAC 991.43/AACC 32.07.01 - Automated Prepare for Test IDF Digestion/Filtration – SDF Precipitation/Filtration If pH check was selected, check and adjust pH as needed 8 Follow Screen Prompts7 IDF SDF Weigh Diatomaceous Earth 3 Weigh Sample4 Insert DE into SDF Bag5 Insert Sample in IDF Bag6 Weigh Filter Bags1 Protein Correction14Ash Correction13Acetone Rinse9 Dry and Weigh10 Acetone Rinse11Dry and Weigh12Insert Filter Bags2

9. Video Demonstration

10. Sample SampleIDF SDF SDFIDF Standard Dev. SDF Cereal7.82.7 7.62.3 7.72.70.100.23 Carrots15.86.4 15.36.4 15.65.6 15.75.8 15.76.5 15.25.90.240.38 Onions11.36.1 11.35.6 11.65.8 11.05.9 11.25.6 11.35.00.190.38 Oat Flakes #14.9 5.20.000.21 Oat Flakes #25.24.9 5.44.60.140.21 Oat Groats8.55.5 8.65.20.070.21 Dietary Fiber - Reproducibility

11. Dietary Fiber – Comparative – TDF SampleReported TDFANKOM TDF Psyllium Husk77.086.4 Cookies5.95.4 Beans6.6 Rice Milk Powder1.91.4 Cereal (Australia)7.58.6 Shrimp0.70.5 Ricies>1.01.4 Rice Milk Powder1.91.4 Wheat Maamoul4.45.2 Blood Sugar Support15.515.7 Memory Support (herbal)17.918.2 Mini Mexican2.11.3 Mini Vegetable2.11.7 Whole wheat pancakes2.9 Veggie Chips7.48.6 Green Seaweed62.561.1 Fiber Supplement (FBIO)85.084.7

12. Sample ID Description Conventional Method Reported Results ANKOM Automated Method Results 8126 Coarse BranCorn Bran76.176.4 8127 Coarse BranCorn Bran78.178.3 8010 Fine BranCorn Bran52.853.0 8012 Fine BranCorn Bran70.370.4 8013 Fine BranCorn Bran54.0 1 61.8 Dietary Fiber – Comparative – IDF/SDF 1 laboratory performing conventional method indicated they had a problem with this sample and were not confident with the reported value

13. AACC Check Samples Check Sample Reported Values IDF ANKOM IDF Reported Values SDF ANKOM SDF AACC DF2 (2012)10.4610.601.751.40 AACC DF3 (2012)13.7913.802.662.60 AACC DF4 (2012)10.009.101.751.00 AACC DF5 (2012)6.515.603.623.10 AACC DF6 (2012)13.5213.302.362.40 AACC DF1 (2013)10.4610.61.751.4 AACC DF2 (2013)6.255.53.894.5 NOTE: The AACC “reported values” are the mean of all reported values, with outliers removed. Twenty-four laboratories participate

14. Cost Comparison Manual Method (per sample)ANKOM (per sample) $115.00$55.00 NOTES: Manual method labor increases with additional samples Labor cost based on $15 (12 €) per hour Did not include labor time to perform Manual method set-up, which could add significant time Lab in Australia indicated one technician was tied up for two days to perform the same amount of samples she could do in one day on the ANKOM system (with free time available).

15.  AOAC 991.43/AACC 32.07.01  Integrated TDF/Resistant Starch &NDO  Upgraded program supports AOAC 2009.01 and 2011.25  Still requires end user HPLC  Maintains AOAC 991.43 option Supported Methods

16. DIETARY FIBER ANALYSIS AUTOMATION & INNOVATION "Dietary fiber is the edible parts of plants or analogous carbohydrates that are resistant to digestion and absorption in the human small intestine with complete or partial fermentation in the large intestine. Dietary fiber includes polysaccharides, oligosaccharides, lignin, and associated plants substances. Dietary fibers promote beneficial physiological effects including laxation, and/or blood cholesterol attenuation, and/or blood glucose attenuation.“ (AACC)

17. Dietary fiber means carbohydrate polymers with ten or more monomeric units, “ which are not hydrolyzed by the endogenous enzymes in the small intestine of humans and belong to the following categories: · Edible carbohydrate polymers naturally occurring in the food as consumed · Carbohydrate polymers, which have been obtained from food raw material by physical, enzymatic or chemical means and which have been shown to have a physiological effect of benefit to health as demonstrated by generally accepted scientific evidence to competent authorities · Synthetic carbohydrate polymers which have been shown to have a physiological effect of benefit to health as demonstrated by generally accepted scientific evidence to competent authorities (CODEX ALIMENTARIUS)

18. Comparison of Dietary Fiber Methods

19. AOAC 991.43 does not capture 100% of the non- digestible oligosaccharides (NDO). Since these behave physiologically as dietary fiber methods were developed for fructan [and fructo-oligosaccharides (FOS)] (AOAC Methods 997.08 and 999.03), galacto-oligosaccharides (GOS) (AOAC Method 2001.03)9, resistant maltodextrins (RMD) (AOAC Method 2001.03)10 and Polydextrose® (AOAC Method 2000.11)11. Why Integrated Dietary Fiber?

20. Demonstration

21. Sample Preparation  Homogeneity  Pre-Extractions –Fat –Sugar –Dry Samples  Liquid Samples

22. Filter Bags  Filter Bag Design –Beaker & Filter  Proper Weighing –Use of the Faraday Cage  Sample Size  Filter Aid (Diatomaceous Earth)

23. Instrument Set-up/Operation  Power Source/Nitrogen/Drain  Preparing Solutions/Enzymes  Getting Started –Computer Controls –Filter Bag Handling

24. Operation – How it works  Digestion Phases (IDF) –Filter/Rinses  Precipitation (SDF/TDF) –Filter/Rinses  Final Rinses (Acetone) –Seal Bags –Rinse rack

25. Protein/Ash Corrections Sample preparation Kjeldahl Vs. DUMAS Kjeldahl Vs. DUMAS Ash Ash