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Natural Resistant Starch

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									                              Natural Resistant Starch:
                              Health Benefits and In Vitro Measurements
                              ________________________________
                              7th International Food Data Conference:
                              Food Composition and Biodiversity
                              Sao Paulo, SP, Brazil,
                              October 21-24, 2007




E. T. Finocchiaro, Ph.D.,
Nutrition R&D Director,
National Starch Food Innovation
Bridgewater, NJ USA
             Presentation Overview


• Resistant Starch (RS) Background

• Scientific Evidence of Key Health Benefits
  – Fermentation
  – Glycemic Response
  – Emerging Research

• In Vitro Analytical Measurements
  – McCleary Method for RS
  – Englyst Methof for RS
  – AOAC Method for today dietary fiber
              What is Resistant Starch?

The Gastrointestinal System              Resistant Starch
                                    is the sum of starch and
                                        products of starch
                 Stomach
                                   degradation not absorbed
                                     in the small intestine of
                                       healthy individuals.
                 Small Intestine




                 Large Intestine
                                    ** A Physiological Definition **
             Natural Sources of Resistant Starch
Food               Serving Size         Resistant Starch,
                                            (grams)
Navy beans         1/2 cup cooked             9.8
Raw banana         One medium, peeled         4.7

Cold potato        One 2” diameter            3.2
Lentils            1/2 cup cooked             2.5

Hi-maize® 260      1 tsp (4 grams)            2.2

Cold pasta         One cup                    1.9
Pearl barley       1/2 cup cooked             1.6
Oatmeal            One cup cooked             0.7
Wholegrain bread   Two slices                 0.5




   Average natural RS consumption in US = 5 grams
Types of Resistant Starch

 RS Type 1        RS Type 2

     +


 Physically       Resistant
inaccessible      granules


 RS Type 3        RS Type 4




Retrograded      Chemically
   starch       modified starch
                  Tolerance and Comfort
            4
           3.5
            3
           2.5
Response    2          *                                                       0g/d RS
           1.5                                                                 60g/d RS
            1
           0.5
            0
                 movements    bloating    abdominal     consistency
                  per day                    pain


     Subjects showed high tolerance to resistant starch.


             n=19 healthy subjects; 1wk;
             bread and cookies containing 0, 50, 75 or 100g of Hi-maize
             *p<0.05, mean ± SEM, scale 1-6


                                                                Kendall et al, FENS, 2003
Fermentation Related Health Benefits of
 High Amylose Maize Resistant Starch
    Outcomes of Resistant Starch Fermentation

            Fermentation
Insulin                                  Satiety


                                   Colonic
  Glucose
                                   Function

        Lipid                  Colonic
      Metabolism                Tissue
                                Health
                   Prebiotic
Benefits from HA maize Resistant Starch Fermentation

Glycemic Benefits
Reduced glycemic response of a subsequent meal
Increased insulin sensitivity

Weight Benefits
Reduced abdominal fat (animal models)
Increased lipid oxidation / fat burning (1 human clinical trial)
Increased production of satiety hormones GLP-1 and PYY (animal
    models)



Other Benefits
Increased absorption of minerals
Increased immune biomarkers
GLYCEMIC Benefits
                         Acute Glucose and Insulin Response
                                   Clinical Study
Glucose (mmol/L)                                    Insulin (pmol/L)
                          Control bread
 7                                                  400
                          High RS bread
6.5                                                 350

                                                    300
 6
                                                    250
5.5
                                                    200
 5
                                                    150
4.5
                                                    100
 4                                                   50

3.5                                                   0
   0   20   40      60     80     100    120              0    20      40      60       80     100     120
                 Time (min)                                                 Time (min)

                     n=20 healthy subjects
                     RS from Hi-maize, flour replacement basis
                     1 meal (b.fast): 2 slices of bread (approx 70g)
                     ~36g CHO; 2g vs. 9g fibre
                     mean ± SEM; AUC & peak difference p<0.05

                                                                                    GI Testing Inc, Canada
    Key Consumer Benefits from Low Glycemic
     Response of HA maize Resistant Starch



•   Helps balance your energy following a meal

•   Helps to maintain healthy blood sugar

•   Implications for multiple benefits
      •   Diabetes
      •   Heart disease
      •   Weight management
      •   Performance and energy
 Emerging Research

1.   Lipid Oxidation
2.   Satiety Hormone Expression
3.   Prebiotics
4.   Mineral Absorption
5.   Insulin Sensitivity
                                                            Lipid Oxidation
                                                Clinical Study – Short Term Response




                                                                  Meal fat oxidation (g oxidized/g FFM)
                             14                                                                                                             24 h

                             12
                                                         *                                                0.06                 *             6h
   Total fat oxidation (g)




                             10                                                                           0.05

                             8                                                                            0.04

                             6                                                                            0.03

                             4                                                                            0.02                *

                             2                                                                            0.01

                             0                                                                            0.00
                                  0        2.7     5.4                                                            0        5.4
                                      RS in meal (%)                                                             RS in meal (%)

          ↑ total fat oxidation                                                                           ↑ dietary fat oxidation
          (by indirect calorimetry)                                                                       (by labelled tracer)
mean ± SE; * p≤0.003 vs. 0%; RS from Hi-maize                           mean ± SE ; * p≤0.0006; RS from Hi-maize


                                                                                                                   Higgins et al, NM 2004
                                                  Satiety
               High-amylose maize RS2 triggered satiety hormones. Cellulose did not.
•   n=15 rats per group, 4wk, control vs RS from Hi-maize vs cellulose diet
•   Genes different: 3192 p<0.05; 1769 p<0.01 (RS vs control)
•   PYY gene: ↑ expression, x1.3 (RS vs control)
•   Proglucagon (GLP-1) gene: ↑ expression, x 3.2 (RS vs control)
                12                                            Gene Expression –
                     pg/100ul
                                           b                  Clustering Analysis
Plasma
PYY




                                                      up




                                                                                          down
                6


                                a   a
                                        mean
                                        ± SE
                0
                          Ctrl          RS
               300
                     pM




                                                      down




                                                                                          up
                                           b
Plasma
               150
GLP-1
                                        mean
                            a       a   ± SE

                0
                                                             Control              RS
                          Ctrl          RS                   In conjunction with
                                                             Louisiana State University
                                                             NAASO Oct 2006
                                     Prebiotics
        Excellent support of Bididobacterium and “good” bacteria


               10
                9
                      *             *
                8                                             *
                7
                6
log 10 / g wet                                                                  Control Diet
                5
caecal content                                                                  RS Diet
                4
                3
                2
                1
                0
                    Bifido-       Lacto-      E.coli    Total
                    bacteria      bacilli               Coliforms


                    n=12 rats, 4wk, 10% Hi-maize in diet, mean+/-SEM, *p<0.05
                    AOM injection, killed 6h later


                                                                     Le Leu et al, JN 2005
                                        Mineral Absorption
                          70
                                          *             Control Diet
                          60                            RS Diet
                                  *
                          50                              *
            Apparent
           Absorption     40
              (%)                                 *                *
                          30

                          20

                          10

                           0
                                Ca      Mg      Zn      Fe       Cu


RS increased mineral absorption.
Proposed mechanism: ↑ solubility at lower pH, ↑ absorptive area, ↑ blood flow


                  n=8 rats; 20% Hi-maize in diet; p<0.05; 3 wk
                  apparent absorption = intake less faecal excretion.

                                                                   Lopez et al, J Nutr, 2001
                                 New 2007 Research

“Effects of resistant starch on insulin sensitivity in diabetic patients”
                                       `
Zhang et al, Chinese Journal of Preventive Medicine, March 2007, 41(2): 101-104

  40 diabetics consumed 30 grams of Hi-maize resistant starch/day for 4 weeks

  FINDINGS
  – Significantly reduced their Body Mass Index (weight)
  – Significantly reduced their fasting insulin levels, glucose response, fructosamine, total
  cholesterol and triglyceride levels
  – Significantly increased insulin sensitivity
Summary of Health Benefits of
  HA maize Resistant Starch
                       Overview: Nutritional Benefits

•   well tolerated within the body
•   easily fermented, with a possible role for digestive health
•   synergistic action with other fibres
•   used to replace digestible carbohydrates like flour, to lower glucose and
    insulin response
•   have a broader metabolic role for the body, via enhanced insulin
    sensitivity and increased lipid oxidation
•   animal studies indicate RS can impact food intake regulation (satiety),
    mineral absorption, and microbial composition
In Vitro Analytical Methods
  to Quantify RS Content
      Key In vitro Analytical Methods


Method           Measures      Key Advantages


McCleary         RS            •Good research tool
Method           (resistant    •AOAC approved
(AOAC 2002.02)   starch)
Englyst          RS            •Good research tool
Method                         •Measures digestion profiles

AOAC TDF         Total fiber   •Most precise & accurate,
(#991.43)                      particularly in food products
                               •AOAC approved
                           The McCleary Method
                   In Vitro Measurement of Resistant Starch
                                AOAC 2002.02

                        Enzymatic Digestion

                                                     Pancreatic
           Incubation at   37oC       16 hr          α-amylase
                                                 Amyloglucosidase



                 Isolate undigested components



                 Dissolve undigested components



    Quantify RS as glucose from solubilized undigested components

McCleary & Monaghan, J. AOAC Int., 2002, 85 (3): 665-675
                                      The Englyst Method
                    In Vitro Measurement of Resistant Starch
                                 Englyst Method


                         Enzymatic Digestion

                                                              Pepsin

                                                              Pancreatin
        Incubation at 37oC                2 hr
                                                             Invertase
                                                          Amyloglucosidase




          Measure glucose released from digested fraction




Englyst et al., Euro J Clinical Nutr, 1999, 46 Suppl 2, S33-S50 & Englyst et al., Am J Clin Nutr 1999, 69, 448-54
                             The TDF Method

                          Total Dietary Fiber Methods
                            AOAC 985.29 & 991.43

                         Enzymatic Digestion
                                                         Amylase
                                                         Protease
  Incubation at 95 o C
                                                     Amyloglucosidase
                            Filtration



            Filtrate                     Insoluble DF
                                                +
Alcohol Precipitation                     Soluble DF




   Supernatant                                 TDF

           W aste                              AOAC Official Methods of Analysis, 1997, 16 th Ed.
     Key Analytical Issues in Quantifying RS in Food Products

1.    RS is measured as TDF in most TDF assays.

2.    TDF analysis is a very precise and accurate method for measuring the RS content
      of many food products.

3.    Analysis by AOAC RS method 2002.02 shows consistently low and variable RS
      contents in various foods products.

4.    The modified Englyst method (original Englyst method using currently available
      enzyme sources) shows higher results with more consistent RS data (vs AOAC
      2002.02 method)
                  TDF and RS Contents of Hi-maize 260
                         by different methods

            60
            50
            40                                                     RS - AOAC 2002.02
  % as is




            30                                                     RS - modified Englyst
            20                                                     TDF- AOAC 991.43

            10
            0
                              Hi-Maize 260


* Hi-maize 260 is a commercial form of granular Resistant Starch
                   Foods analyzed by Different Methods



                           TDF and RS Contents of Foods
                                with Hi-maize 260


           20

           15
                                                                    RS - AOAC 2002.02
% a s is   10                                                       RS - modified Englyst
                                                                    TDF- AOAC 991.43
           5

           0
                 Breads        Pasta        Tortilla     Biscuits




*Hi-maize 260 is a commercial form of granular Resistant Starch
       Key Take Aways for In Vitro Analytical Methods.



1. RS is measured as TDF in most TDF assays.

2. Claims substantiation studies generally require the analytical
   quantification of fiber (or RS) in the selected clinical food product.

3. TDF assay AOAC 991.43 is the preferred method to quantify
   granular RS in food products.

4. More development work is needed for a comprehensive RS method.
THANK YOU
                   Resistant Starch and Glycaemic Response:
                              Strength of Evidence
                            Significant Decrease          Significant Increase /
                                                                 No Effect

 Glucose response

                                                                                 10/13 +ve


 Insulin response
                                                                                 8/12 +ve



Review of 15 clinical studies (Passclaim level I/II) utilising high amylose maize starch.
Strong evidence supporting effect of RS on short term response.
                                                       Increase / No Effect   Decrease
                  Resistant Starch Fermentation:
                       Strength of Evidence
Review of 12 clinical studies (Passclaim level I/II) utilizing high amylose corn RS2 in
foods. Strong evidence supporting RS fermentation.             Significant Non-significant



                         Decrease                       Increase

Fecal weight                                                                        8/9 +ve


Fecal frequency                                                                     4/4 +ve

Fecal SCFA                                                                          6/6 +ve


Fecal butyrate                                                                      4/5 +ve

Fecal pH                                                                            7/7 -ve
                Recognition of Resistant Starch
Leading nutrition authorities
• Global: World Health Organization, 1997
• USA: Institute of Medicine, National Academies, 2002
• UK: British Nutrition Foundation, 2005
• Australia / NZ: National Health and Medical Research Council, 2006


Research community
• Medline database
  in vitro, animal, clinical studies
                              Insulin Sensitivity
                  Clinical Study – Medium Term Response
12
     mg gluc.kg-1.min-1
       / mU insulin-1
                                    • Increased insulin sensitivity
11
                               *    following a high RS diet.
10
                                    • Associated with lower
9                                   insulin concentration, higher
                                    glucose clearance, lower
8                                   adipose NEFA and higher
                                    adipose SCFA uptake.
7

                                   n=10 healthy subjects; 4wk; 30g RS from Hi-maize
6
                                   *p=0.027; mean ± SEM; measured by H-E clamp
           Control        RS




                                                    Robertson et al, AJCN, 2005

								
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