Hydrotherm Fat Determination Study 1 HYDROTHERM (WEIBULL – STOLDT METHOD) VS. TRADITIONAL AOAC
922.06
• Hydrotherm meets/exceeds the requirements of AOAC method 922.06 • Hydrotherm advantages:
Improved operational efficiency -‐ time savings of nearly 7 minutes per sample tested Cost Reduction of 22% per sample tested Safer work environment – reduced risk of lost time incident
2 UNDERSTANDING FAT • What is Fat?
Triglycerides – organic compounds free of nitrogen Metabolic products found in plants and animals Like proteins and carbohydrates, it is a nutrient used as an energy source by the body Has a caloric value…1g of fat = 9 calories or approximately 38 kilojoules Often referred to as a lipid, it is insoluble in water, but soluble in organic solvents
• Fat is a key component of the foods we eat Nutrition Appearance Texture Flavor
• Fat extraction process Extractable fat is called, “free fat” Fat that is bound or enclosed by other organic components cannot be directly extracted To determine total fat content the sample must be digested prior to extraction
The US Nutrition labeling and Education Act of 1990 (NLEA) has defined total fat as the sum of all fatty acids obtained from a total lipid extract expressed at triglycerides
• Need for fat extraction FDA mandate for food nutritional value label Mandatory label information
Total Fat, Saturated Fat, Trans Fat, Cholesterol Sodium, Calcium, Iron Vitamin A, Vitamin C Total Carbohydrates Dietary Fiber Sugars Protein Total Calories, Calories from Fats
Determination of “Good” and “Bad” fat
• Calculating the correct total fat content Sum parameter defined by the selected test method Determined using the sample matrix Can be further characterized based on fatty acid spectrum
3 STUDY BACKGROUND • Cooperative evaluation between Lab Synergy and Tyson Foods
Evaluation of acid hydrolysis Test Methods used
Traditional AOAC 922.06 method Hydrotherm Weibull-‐Stoldt method
Food and Feed products with various mixed matrices tested Over 1000 data points collected
• Standard AOAC 922.06 Acid Hydrolysis Test Method 1.) 2 g. samples to 50 mL beaker…add 10 mL HCI…mix well, place in hot water bath for 35
minutes…add 10 mL alcohol and cool. 2.) Transfer aforementioned mixture to Mojonnier fat-‐extraction apparatus. Add in 3 portions
to flask with stopper, shake vigorously...add 25 mL redistilled petroleum ether and again shake vigorously…let stand until upper liquid is practically clear, or centrifuge 20 minutes at ca 600 ppm.
3.) Draw off ether-‐fat solution through filter into 125 mL beaker-‐flask containing porcelain chips or broken glass
4.) Re-‐extract liquid remaining in tube twice, each time with only 15 mL of each ether...shake well on addition of each ether…draw off clear ether solutions through filter into same flask as before…evaporate either slowly on steam bath…dry fat in oven at 100°C to constant weight…remove flask and counterpoise from oven, let stand in air to contain weight, and weigh…correct this weight by blank determination on reagents used…report as % fat by acid hydrolysis.
• Weibull-‐Stoldt Method
1.) Hydrolysis of sample with HCI using the Hydrotherm closed loop automated system
Filtration Drying of filter
2.) Extraction of fats from filtration residue by use of a single organic solvent – performed using the Soxtherm closed loop automated extraction system
Evaporation of the solvent Drying of the fat
3.) Weighing the fat 4.) Summary/Benefits
Repeatable Precise results Universal use High through-‐put Improved operating efficiency Lower cost No fume hood required
• Hydrotherm (Weibull-‐Stold) vs. AOAC 922.06 Comparative Data
Stats Hydrotherm Observations
Overall Testing Cost Savings Time Savings Safety Advantages
Conclusion