presentation functional food group 3
TRANSCRIPT
Scientific Opinion on the substantiation of a health claim related to barley soup “Orzotto” and protection of blood lipids from oxidative damage
Presented by:
Jaturong Amonchaisup
Sophia Amenyah
Rafat KarbakhshPriscille Giner
Introduction / Background • The Applicant: Italsur srl, Italy• Product Name: “Orzotto”• Health Claim :“Orzotto contributes to protection of blood lipids from oxidative damage”. Daily inclusion of a portion of frozen barley soup “Orzotto” in a balanced and varied diet reduces oxidative damage of blood lipids.
• Target group: The general population
• Amount & Duration of consumption: 250 g barley soup “Orzotto” daily for two weeks.
Pearled Barley
Tri-colored Swiss Chard Tuscan cabbage Bi-colored Spinach Savoy Cabbage
Purple carrotPotatoRed/Yellow Celery
Parsley onion
75% Pearled Barley
2.5%
2.5%
2.5%
2.5%
2.5%2.5%
2.5%2.5% 2.5% 2.5%
Pearled Barley
Tuscan black cabbage
Potatoes
Blue savoy cabbage
Purple carrot
Bi-colored Spinach
Tri-colored Swiss Chard
Onion
Yellow Celery
Red Celery
Parsley
Composition of Orzotto250 g Orzotto
• Health Claim: Contributes to the protection of blood lipids from oxidative damage
• Need to know:1) Mechanism of oxidative damage of lipid2) Bioactive compounds and their
mechanisms in preventing lipid oxidation.
• Potential bioactive compound: Antioxidants and radical scavenger.
• Major: Phenolic coumpounds• Minor: Tocopherol, Carotenoid, Ascorbic
acid, Selenium But, present in marginal amount
Phenolic compounds as antioxidant and radical
scavenger
1) Radical scavenging activity - Give H atom to peroxide radical - Peroxide radical Hydro peroxide (non-radical) - Phenolics Phenolic radicals (non-reactive)
Phenolic compounds as antioxidant and radical scavenger
2) Metal Sequestrant
Phenolics chelate with metal so metal become unavailable to initiate lipid oxidation
Total Phenolic compounds CompoundsIngredient
Proportion in Orzotto
(%)
Amount in Orzotto
(g/ 250 g)Major type of
phenolic compounds
Average Total Phenolic
compounds(mg / 100 g FW)
Amount Phenolic compounds in each
ingredient
Total Phenolic compounds in
food(mg/250 g)
Pearled Barley 75% 187.5 Phenolic Acids,
Proanthocyanidins 763.00 1,430.63
1,569
Tuscan black cabbage 2.5% 6.25 Quercetin, Phenolic
Acid 831.00 51.94
Potatoes 2.5% 6.25 Chlorogenic acid,
anthocyanins 33.33 2.08
Blue savoy cabbage 2.5% 6.25 Flavonoids 50.97 3.19
Purple carrot 2.5% 6.25
Anthocyanin, Hydroxycinnamic acid, syringic acids
74.64 4.67
Bi-colored Spinach 2.5% 6.25 Flavonoid: Quertecin,
Anthocyanin 278.35 17.40
Tri-colored Swiss Chard 2.5% 6.25
Flavonoid: Catechins, Kaempferol, Quertecin
192.00 12.00
Onion
2.5% 6.25Flavonoid: Quercetin, Isorhamnetin, Myricetin and Kaempferol
645.00 40.31
Celery 5% 12.5 Furanocoumarins,
bergapten, Apigenin 4.49 0.56
Parsley 2.5% 6.25 Flavonoids, Apigenin 103.70 6.48
Effect of Processing on Total Phenolic Content
Total phenolic compounds in 250 g Orzotto
(mg)
Loss during Process(Freezing + Thermal
process)
Remaining amount of phenolic compounds in 250 g Orzotto
(mg)
1,569 20 – 30 % 1,100
• Freezing– Causes minimal destruction – Sometimes, increases in the phenolic content
• Heating prior consumption– Few minutes minimal destruction – Sometimes, increases in the phenolic content
• Leaching– Major cause of loss– But, leached nutrients are still present in soup matrix
Total Ascorbic AcidIngredient
Proportion in Orzotto
(%)
Amount in Orzotto
(g/ 250 g)
Average Ascorbic acid after cooking process
(mg / 100 g FW)
Amount of Ascorbic acid in each ingredient
Total Ascorbic acid
in food(mg/250 g)
Pearled Barley 75% 187.5 0 0.00
16.79
Tuscan black cabbage 2.5% 6.25 41 2.56
Potatoes 2.5% 6.25 13 0.81Blue savoy cabbage 2.5% 6.25 17 1.06
Purple carrot 2.5% 6.25 3.6 0.23Bi-colored Spinach 2.5% 6.25 9.8 0.61
Tri-colored Swiss Chard 2.5% 6.25 18 1.13
Onion 2.5% 6.25 5.2 0.33Celery 5% 12.5 6.1 0.76Parsley 2.5% 6.25 149 9.31
Total TocopherolIngredient
Proportion in Orzotto
(%)
Amount in Orzotto
(g/ 250 g)
Average Ascorbic acid after cooking process
(mg / 100 g FW)
Amount of Ascorbic acid in each ingredient
Total Ascorbic acid
in food(mg/250 g)
Pearled Barley 75% 187.5 0.01 0.02
1.19
Tuscan black cabbage 2.5% 6.25 0.85 0.05
Potatoes 2.5% 6.25 0.01 0.00Blue savoy cabbage 2.5% 6.25 0 0.00
Purple carrot 2.5% 6.25 1.03 0.06Bi-colored Spinach 2.5% 6.25 2.08 0.13
Tri-colored Swiss Chard 2.5% 6.25 1.89 0.12
Onion 2.5% 6.25 0 0.00Celery 5% 12.5 6.1 0.76Parsley 2.5% 6.25 0.7 0.04
Total SeleniumIngredient
Proportion in Orzotto
(%)
Amount in Orzotto
(g/ 250 g)
Average Ascorbic acid after cooking process
(µg / 100 g FW)
Amount of Ascorbic acid in each ingredient
Total Ascorbic acid
in food(µg/250 g)
Pearled Barley 75% 187.5 8.6 16.13
18.9
Tuscan black cabbage 2.5% 6.25 0.9 0.06
Potatoes 2.5% 6.25 0.3 0.02Blue savoy cabbage 2.5% 6.25 0.7 0.04
Purple carrot 2.5% 6.25 0.7 0.04Bi-colored Spinach 2.5% 6.25 6 0.38
Tri-colored Swiss Chard 2.5% 6.25 0.9 0.06
Onion 2.5% 6.25 0.6 0.04Celery 5% 12.5 1.0 0.13Parsley 2.5% 6.25 32.3 2.02
Total Amount of Bioactive Compound
250 g Orzotto
Bioactive Compound Amount
Phenolic compounds 1,100 mg
Ascorbic acid 16.79 mg
Tocopherol 1.19 mg
Selenium 18.9 µg
Human Clinical Studies:- Study proposed by applicant is unpublished Need a literature search on the effect of phenolic compounds in protecting blood lipid against oxidative damage.
- Literature search on PubMed, Cochrane Library, Web of knowledge and Google scholar
- Several RCT and clinical trials with significantly positive results were found.
Biomarkers used in the assessment of lipid oxidation
Direct measurement (based on EFSA documents)
• Decreased plasma oxidized LDL
• Decreased F2-isoprostanes (product from the peroxidation of essential fatty acid primarily arachidonic acid )
Indirect measurement (based on EFSA documents)
• Increased lag time before initiation of LDL oxidation
• Decreased plasma TBARS (Thiobarbituric acid reactive substances = by product of lipid peroxidation)
• Decreased plasma malondialdehyde (product of lipid peroxidation)
Phenolic compounds (>500 mg/day)Food Intake Quantity Duration Subjects Results
Pomegranate juice
570 – 1000 mg/day total
phenolic compounds
(mainly Flavonoids)
Average:2 weeks
Average: 13 healthy
subjects
Significant decrease in• Plasma oxidized LDL• F2-isoprostanes• Plasma TBARS• Lipid peroxide
Significant increase in• lag time before the initiation of LDL oxidation
Sweet potato leaves
Perilla
Tart cherry juice
Phenolic compounds (400-500mg/day)Food Intake Quantity Duration Subjects Results
Black tea
Avg. 460 mg/day total phenolic compounds
(mainly Flavonoids esp.
catechin)
2 – 8 weeks Average: 30 healthy
adults
Significant decrease in• Plasma oxidized LDL• Plasma MDA• Lipid peroxide
Significant increase in• lag time before the initiation of LDL oxidation
cranberry juice
Green tea polyphenol
extract
Catechin
Phenolic compounds (<400 mg/day)Food Intake Quantity Duration Subjects Results
Soy germ pasta
< 400 mg/day total phenolic compounds
(mainly Flavonoids)
Average:8 weeks
Average: 20 healthy
subjects
Significant decrease in• Plasma oxidized LDL• F2-isoprostanes• Lipid peroxide
Pycnogenol (Extract)
Cocoa powder
Grape-seedextract
According to these studies,
• Significant results in those clinical studies + Consistency of the results with similar studies carried out on the same food.
• The type of phenolic compounds that are available in our product are similar to those from studies (flavonoids such as quercetin, Proanthocyanidins and anthocyanin)
• And also they are all plant based product, so we expect to see more or less the same bioavailability.
• Our product provides 1,100 mg phenolic compound in each time intake, which is higher than those present in the most studies. So, we expect a good effect after 2 week of daily consumption.
• Also our product also provide 16.79 mg Vitamin C, 1.19 Vitamin E and 18.9 µg Selenium, which would have helping role in addition to the phenolic compounds.
Literatures which show that each vegetable ingredient can protect blood lipid against oxidative damage.
Vegetables Results and conclusion of the study Type of study
Pearled Barley Pearled Barley shows inhibitory effects toward human LDL oxidation
In vitro
Potatoes Metanolic extract of potato is able to inhibit linoleic acid oxidation
In vitro
Tri-colored swiss chard The aorta lipid peroxidation was found to be decreased significantly
Animal study (Rat) - In vivo
Onion Onion is able to inhibit LDL oxidation and increase the resistance of LDL against oxidation in vitro
In vitro
Celery Decrease of liposomal peroxidation intensity in liver and blood of mice
Animal study (Mice) - In vivo
Parsley Reduction of the intensity of lipid peroxidation in mice liver and blood
Animal study (Mice) - In vivo
Vegetable Results and conclusion of the study Type of study
Bi-colored spinach
Spinach is able to influence lipid peroxidation in a beneficial manner
intervention study in healthy human
Blue savoy cabbage
Phenolic extracts from Blue savoy cabbage inhibit a lipid peroxidation in a linoleic acid
emulsion
In vitro
Tuscan Black cabbage
Tuscan Black cabbage shows a protective effect on the oxidation of lipoproteins
In vitro
Purpule Carrot
Drinking carrot juice decreases lipid peroxidation
Human study – In vivo
Soup ingredients Type of study
Quantity of soup intake
Duration of intervention
Results
Barley soup (Orzotto) Human study (single-arm)
250 g/d 2 weeks
Serum levels of oxidised LDL
decreased after consumption
2 kinds of soup with carrot, tomato, broccoli and olive oil
Human study - clinical control trial
300 mL/d 4 weeks
Gazpacho (80% crude vegetables : tomato, cucumber, pepper ; olive oil and other minor components)
Human study - clinical control trial
500 mL/d 14 days
Literatures which show that the similar vegetable soups can protect blood lipid against oxidative damage
Evaluation based on Criteria for scientific substantiationCriteria
1) The food or food component should be characterized
2) Substantiation of a claim should be based on human studies
2a) Study groups are representative of the target group
2b) Appropriate controls
2c) An adequate duration of exposure and follow up to demonstrate the intended effect
2d) Characterization of the study groups’ background diet and other relevant aspects of lifestyle
2e) An amount of the food consistent with its intended pattern of consumption
2f) The influence of the food matrix on the functional effect of food
2g) Monitoring of subjects’ compliance concerning intake of food
3) Markers are used when the true endpoint cannot be measured
4) Markers should be biologically and methodologically valid
5) A Change in target variable is statistically different and biologically meaningful
6) Taking into account the totality of the available data and by weighing of the evidence.
Synergistic effect
Phenolics Phenolic radical
Apart from phenolic compounds, the food product also contain vitamin C
- Phenolics give H atom to radical - Radicals Non-radicals- Phenolics Phenolic radicals- Ascorbic acid give H atom to Phenolic radicals- Ascorbic acid Dehydroascorbic acid- Phenolic radicals Phenolics
Phenolic compounds (<400 mg/day)
Food Quantity of phenolic compound
intake
Duration of Intervention
Biomarkers Subjects Type of study
soy germ pasta
31–33 mg/day isoflavones 8 weeks Decreased plasma
oxidized LDL
26 adults with T2D (13 men/13 women, aged 62 ± 7 years)
RCT
Pycnogenol (Extract)
150 mg/day proanthocyanidins 8 weeks Decreased F2-
isoprostanes
23 patients with coronary artery disease (CAD)
RCT
Cocoa drink 187 mg /day Flavanols < 1 day Decreased F2-
isoprostanes 20 Healthy subjects RCT
Grape-seedextract
300 mg/day total phenolic < 1 day - Decreased lipid
peroxide 8 Healthy subjects RCT
Phenolic compounds (400-500mg/day)
Food Intake Quantity Duration of Intervention
Biomarkers Subjects Type of study
Black tea
400 mg/day flavonoid(Catechins and theaflavins)
4 weeks
lag time before the initiation of LDL oxidation(Prolonged by 8 min (10-15%))
healthy male volunteers (n = 22) aged 22 ± 1 y(14 treatment, 18 control)
Clinical Control trial
cranberry juice
460 mg/day total phenolic 8 weeks
- Decreased plasma oxidized LDL - Decreased plasma malondialdehyde
Adults (n = 36) with features of metabolic syndrome
RCT
Green tea polyphenol
extract
480 mg/day catechin 2 weeks
lag time before the initiation of LDL oxidation(prolonged by 14 min)
22 male volunteers aged between 22 and 32 years
Clinical control trial
Catechin 500 mg/day catechin 4 weeks Decreased plasma
oxidized LDL
- 40 healthy adult volunteers (10 men, 30 women) - catechin group (n = 29)
Clinical control trial
Phenolic compounds (>500 mg/day)Food Intake Quantity Duration of
InterventionBiomarkers Subjects Type of
study
Pomegranate juice
570 mg/daytotal phenolics
until disease progression
Decreased lipid peroxide - Clinical
trial
Sweet potato leaves
902 mg/day polyphenols 2 weeks Decreased plasma
TBARS15 healthy male volunteers RCT
High purple sweet potato
leaves
902 mg/day polyphenols 2 weeks
- Prolonged lag time before the initiation of LDL oxidation
15 elite basketball players (6 men and 9 women, aged 20-24 y)
RCT
Perilla 1000 mg polyphenol/day 4 hr
- Prolonged lag time before the initiation of LDL oxidation - Decreased lipid peroxide - TBARS
Female healthy volunteers (n = 8)
RCT
Tart cherry juice
1100 mg polyphenol/day 2 weeks Decreased F2-
isoprostanes
12 healthy men and women (6 men, 6 women) aged 61–75 y
RCT
Literatures which show that each vegetable ingredient can protect blood lipid against oxidative damage.
Vegetable
Method Results and conclusion of the
study
Type of study
Amount of vegetables used
in the experiment
Pearled Barley
Difference between the amount of formed diene in the control sample (antioxidant free) and in the presence of antioxidant barley extracts
Pearled Bareley shows inhibitory effects toward human LDL oxidation
In vitro Extracts prepared by weighing 1,0g wholegrain
Potatoes Antioxidant activities of potato extracts compared with a commercially available antioxidant (α-tocopherol)
Metanolic extract of potato is able to inhibit linoleic acid oxidation
In vitro total phenolic: 16,58 to 36,24 mg GAE/100g dry sample
Tri-colored swiss chard
Chard extract added in rat diet compared to a basic diet without chard
The aorta lipid peroxidation was found to be decreased significantly
Animal study (Rat) - In vivo
Chard extract (2g/kg) in 28 days
Vegetable Method Results and conclusion of the study
Type of study
Amount of vegetables used in the experiment
Onion Inhibitory effect of extracts of onion on LDL oxidation in vitro compared with shallot and garlic extracts
Onion is able to inhibit LDL oxidation and increase the resistance of LDL against oxidation in vitro
In vitro Onion concentration: 5 and 10 mg/mL
Celery Treatment with Celery leaves extracts compared to a control (no treatment)
Decrease of liposomal peroxidation intensity in liver and blood of mice
Animal study (Mice) - In vivo
1.0 mL/kg (i.p.) of celery leaves extracts at the same time.
Parsley Treatment with Parsley leaves extracts compared to a control (no treatment)
Reduction of the intensity of lipid peroxidation in mice liver and blood
Animal study (Mice) - In vivo
1.0 mL/kg (i.p.) of parsley
Purpule Carrot
Effects of drinking carrot juice on anthropometrics, blood Pressure, blood chemistry, lipid panel, hormones, inflammatory markers, and antioxidant status before and after the test
Drinking carrot juice decreases lipid peroxidation
Human study – In vivo
16 fl oz of daily freshly squeezed carrot juice for 3 months
Vegetable
Method Results and conclusion of the
study
Type of study
Amount of vegetables used
in the experiment
Bi-colored spinach
Comparaison between markers of lipid peroxidation
before and after a 10 days intake of spinach
Spinach is able to influence lipid
peroxidation in a beneficial manner
intervention study in healthy
individuals
6,2 g of spinach power contains
5mg/lutein
Blue savoy cabbage
Evaluation of the antioxidant capacity and content of Blue
savoye cabbage
Phenolic extracts from Blue savoye cabbage inhibit a
lipid peroxidation in a linoleic acid emulsion
In vitro2g and 0,5g/L of
linoleic acid emulsion
Tuscan Black
cabbage
Comparaison beteween lipid peroxidation levels of
oxidized lipoproteins without and with vegetable extracts at
different concentrations
Tuscan Black cabbage shows a protective
effect on the oxidation of lipoproteins
In vitro 5/10/25/50 ug/mL
Soup ingredients Type of study
Quantity of soup intake
Duration of intervention
Results
Barley soup (Orzotto) Human study (single-arm)
250 g/d 2 weeks Serum levels of oxidised LDL decreased after consumption
2 kinds of soup with carrot, tomato, broccoli and olive oil
Human study - clinical control trial
300 mL/d 4 weeks Serum levels of oxidised LDL decreased after consumption of both soups
Gazpacho (80% crude vegetables : tomato, cucumber, pepper ; olive oil and other minor components)
Human study - clinical control trial
500 mL/d 14 days Increase of serum vitamin C concentration
Literatures which show that the similar vegetable soups can protect blood lipid against oxidative damage