Analysis of Total Fat

Total Fat
Refers to the sum of triglycerides, phospholipids, wax ester, sterols and minor amount of non-fatty materials

Methods
Sum of individual triglyceride X Gravimetric methods Common techniques Soxhlet extraction (without hydrolysis)
Acid / alkaline hydrolysis

Soxhlet extraction Vs Acid hydrolysis

GB/T 5009.6-2003 Determination of fat in foods Soxhlet extraction
Crude fat including free fat () Acid hydrolysis Total fat including free and bounded fat ()
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AOAC official methods

Acid hydrolysis
Method 963.15 925.12 945.44 922.06 935.38* 925.32* 948.15* 948.16 950.54 Cacao products Macaroni Product Fig bars & raisin-filled crackers Flour bread eggs Seafood Fish meal Food dressings Matrix Solvent Petroleum ether Ether / petroleum ether Ether / petroleum ether Ether / petroleum ether Ether / petroleum ether Ether / petroleum ether Ether / petroleum ether Acetone Ether / petroleum ether

* Procedure similar to AOAC 922.06

AOAC official methods

Alkaline hydrolysis
Method 989.05 920.111* 995.19 945.48* 920.115* 922.09* 932.06* 986.25* 933.05* 974.09* 952.06* Milk Cream Cream Evaporated and condensed milk Sweet condensed milk Malted milk Milk powder Milk-based infant formula Cheese Whey cheese Ice cream and frozen desserts Matrix Solvent Ether / petroleum ether Ether / petroleum ether Ether / petroleum ether Ether / petroleum ether Ether / petroleum ether Ether / petroleum ether Ether / petroleum ether Ether / petroleum ether Ether / petroleum ether Ether / petroleum ether Ether / petroleum ether 6

* Procedure similar to AOAC 989.05

AOAC official methods

Without hydrolysis (Soxhlet extraction)

Method 960.39 991.36 948.22 Meat

Matrix

Solvent Anhydrous ether or petroleum ether Petroleum ether Ether (16h)

Meat and meat products Nuts and nut products

AOAC official methods

Other techniques
Method 938.06 995.18 989.04 2000.18 960.26 969.16 973.22 972.16 969.24 964.12 976.21 985.15 Butter Cream Raw milk Raw, whole milk Raw milk milk milk milk Fish meal Seafood Meat Meat and poultry products Matrix Techniques Wash sample with ether or petroleum ether, fat = 100 water -residue Babcock method Babcock method Gerber method Rapid detergent method Automated turbidimetric method Automated turbidimetric method Mid-infrared spectroscopic method Semimicro method, extraction apparatus Rapid modified babcock method Rapid specific gravity method Rapid microwave solvent extraction C2Cl4 CH2Cl2 8 CHCl3 Solvent

Points to note
Soxhlet extraction dried samples is required results may vary with the reflux rate and extraction time Acid hydrolysis can produce higher results for cereal products

Flow chart for acid hydrolysis

Homogenous Homogenous Sample Sample Acid Acidhydrolysis hydrolysis Solvent Solventextraction extraction

Total Totalfat fat (weight (weightof ofresidue) residue)

Evaporate Evaporatesolvent solvent

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Sample preparation
edible and non-edible portions water content of sample representative and homogenous sample sample size => Definition of 0 0.5 g/100g e.g. if sample contains 0.5% fat, 1 g sample contains (1 x 0.5%) = 0.005 g of fat
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Blending

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Drying
Freeze-dryer

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Acid hydrolysis
Reagent: ~ 8.3 M HCl Temp: 70 - 80 C Time: 30 - 40 min

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Extraction
Solvents: diethyl ether and petroleum ether

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Weighing
Determine the weight of residue
Drying to constant weight Prolong heating may increase weight of fat, due to oxidation

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Available proficiency test

FAPAS AOAC LGC AOCS

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CRM
NIST BCR LGC/ERM

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Thank You

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