Analysis of Dietary Fibre
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1 Analysis of Dietary Fibre 1
2 HK Regulation Dietary fibre means any fibre analyzed by means of any official methods adopted by AOAC International. 2
3 AOAC Official Methods AOAC Total Dietary Fibre in Foods Enzymatic-Gravimetric Method AOAC AOAC AOAC AOAC etc Soluble Dietary Fibre in Food and Food Products Insoluble Dietary Fibre in Foods and Food Products Total, Soluble, and Insoluble Deitary Fibre in Foods Total Dietary Fibre (Applicable to determination of total fibre in cereals, beans, vegetables and fruits) Enzymatic-Gravimetric Method Enzymatic-Gravimetric Method Enzymatic-Gravimetric Method Enzymatic-Gravimetric Method 3
4 AOAC Duplicate test test portions of of sample Enzymatic digestion i) i) α-amylase ii) ii) protease iii) iii) amyloglucosidase Precipitation and and filtration (residue) One One residue for for protein analysis One One residue for for ash ash analysis Total dietary fibre = weight (residue) weight (ash + protein) 4
5 AOAC Homogenize and dry sample (freeze-dry is recommended). Defat with petroleum ether if >10% fat content, otherwise false high results. Weigh duplicate test portions (difference in weight should not >20 mg). 5
6 AOAC Preparing for digestion 6
7 AOAC Preparing for digestion 7
8 AOAC α-amylase: gelatinize Add phosphate buffer (ph 6.0, 50 ml) to sample. Adjust to ph 6.0 ± 0.2. Add enzyme, incubate at C. 30 minutes in water bath. 8
9 AOAC Protease: remove protein Cool to room temperature. Adjust to ph 7.5 ± 0.2. Add enzyme, incubate at 60 C for 30 minutes. 9
10 AOAC Amyloglucosidase: remove starch Cool. Adjust to ph Add enzyme, incubate at 60 C for 30 minutes. Note: final solution volume is about 70 ml. 10
11 AOAC Summary ph 6.0 ± 0.2, α-amylase, C, min. ph 7.5 ± 0.2, protease, 60 C, 30 min. ph , amyloglucosidase, 60 C, 30 min. 11
12 AOAC Add 280 ml 95 % ethyl alcohol (60 o C). Let precipitate form at room temperature for 60 minutes. Collect the residues (soluble fibre + insoluble fibre) in pre-weight crucibles. 12
13 AOAC Precipitation 13
14 AOAC Filtration 14
15 AOAC One test portion for protein, using N x 6.25 as conversion factor. Incinerate second test portion at 525 o C for 5 hours. Total dietary fibre = [weight residue protein ash blank] / weight test portion Weight residue = average of duplicate Weight test portion = average of duplicate 15
16 AOAC Test for enzyme purity every half yearly Test sample Activity tested Test portion (g) Expected recovery (%) Citrus pectin Pectinase Stractan (larch gum) Hemicellulase Wheat Starch Amylase Corn Starch Amylase Casein Protease β-glucan (barley gum) β-glucanase
17 AOAC Method for functional fibre Functional fibre Beta-glucan Oligofructose Fructooligosaccharides Polydextrose Galactooligosaccharides Resistant maltodextrin Resistant starch Commercial name Imprime PGG Raftilose, OliggoFiber TM Neosugar, Actilght Litesse Yacult, Borculo Whey Products Fibersol-2 C*Actistar Test Method AOAC AOAC or AOAC or AOAC AOAC AOAC AOAC
18 AOAC Dietary Fiber Containing Supplemented Resistant Maltodextrin (RMD) High MW RMD by Method and Low MW RMD by HPLC 18
19 AOAC Filtrate (by (by ) i) i) Concentrate ii) ii) Desalt Analysed by by LC LC Total dietary fibre = [weight of of residue -- protein ash --blank] + [weight of of low low MW soluble fibre determined by by HPLC] 19
20 AOAC Only a portion of the resistant maltodextrin is precipitated in the aqueous ethanol when Method is applied to foods containing resistant maltodextrin. 20
21 AOAC In Method , resistant maltodextrin that are soluble in aqueous ethanol are desalted, concentrated and measured by liquid chromatography. 21
22 Desalting Ion exchange column (OH-type and H-type) 22
23 AOAC Total dietary fibre = fibre as measured by Method plus low molecular weight resistant maltodextrin Note: low molecular weight resistant maltodextrin refers to resistant maltodextrin that are soluble in the aqueous ethanol in Method
24 Calculation of available carbohydrate when method is used Available carbohydrate = 100 [moisture + ash + protein + fat + alcohol + (soluble dietary fibre + insoluble dietary fibre) + low molecular weight resistant maltodextrin] Note: low molecular weight resistant maltodextrin refers to resistant maltodextrin that is measured by AOAC
25 Recoveries for other functional fibres when tested by AOAC Functional fibre Recovery (%) β-glucan Fructooligosaccharides Polydextrose Galactooligosaccharides Glucooligosaccharides Resistant maltodextrin 101* * Recovery of insoluble dietary fibre and soluble dietary fibre is particular high. 25
26 Points to note Definition of 0 : 1 g/100 g. Results are method dependent. Functional fibre should be included in the calculation of available carbohydrate. AOAC may give higher TDF results but with higher testing cost. 26
27 Available proficiency test FAPAS AOAC LGC 27
28 Thank You 28
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Type 2 Diabetes What is Type 2 Diabetes? Diabetes is a condition where there is too much glucose in the blood. Our pancreas produces a hormone called insulin. Insulin works to regulate our blood glucose