FINAL REPORT

September 2015

CENTRE D’ETUDE ET DE VALORISATION DES ALGUES PRESQU’ILE DE PEN-LAN – B.P.3 – F- 22610 PLEUBIAN TEL : +33 (0) 2 96 229.350 – FAX : +33 (0)2 96 228 438

ALGUE@CEVA.FR   — WWW.CEVA.FR

1.       Recall

VitaPro is a company selling a patented formulation of pharmaceutical grade apple pectin as a nutritional supplement for detoxification, cholesterol management and blood glucose management applications.

Centre d’Etude et de Valorisation des Algues (CEVA) was established in 1982. It provides to its customers expertise, information centre, laboratories and pilot plant facilities for studies, and development of innovating products.

2.       Work programme

2.1.       Proposed study – Detoxification Properties of apple pectin extract

The aim of this study is to quantify the chelating effect of ProPectin on selected heavy metals, known as toxic for human health.

A known amount of apple pectin extract will be set in chemical conditions mimicking human digestion (see below). A mixture of heavy metals (Cd, Sn Hg, Pb) will be added to the solution at concentrations above the average security limit defined for daily food intake.

Samples will be analysed for individual metal concentrations using ICP-OES techniques. Sampling will be performed along the digestion process, as described above. The experiment will run at the same time as Part 1, using same samples.

In case of high detox properties, metals will be chelated by pectin and will not pass through the UF membrane. In case of weak detox properties, metals will pass the membrane. Potential hydrolysis of pectin may impact detox properties.

3.       Results

3.1.       Sampling Strategy

• Initial raw product tested in our experiments :

• Step 1 : Solutions before digestion experiments
  • Solutions of pectin samples dissolved in water = T0
• Step 2 : Gastric digestion (pH 2, 37°C, with pepsin)
  • After 1 H at 37°C : sampling of the solution = T1
• Step 3 : Partial intestinal digestion (pH 7.5)
  • 2 H at 37°C, then 1 H at 4°C to stop process
  • Centrifugation of the solution,
    • sampling of supernatant = T2
    • Sampling of solid phase (if any) = “culot”
• Step 4 : Simulation of intestinal absorption
  • Filtration trough Ultra-Filtration membrane set at 3 000 Da (3 kDa).
  • Retentat = T3
  • Permeat = T4

3.2.       Part O – Analysis of ProPectin

Table 1 : Analytical results ProPectin. All concentrations are expressed in %/raw = % in original product as it is delivered.

Results discussion

• Natural Apple pectins are made of a mixture of polysaccharides rich in Galacturonic acid (Gal acid). Composition may vary according to apple species, season and extraction procedures, however a pure natural apple pectin is always rich in Galacturonic acid.
• The VitaPro ProPectin is an optimized formulation containing 27 % of  pure  pharmaceutical grade apple pectin and natural fructose. The profile of oses, galactose, galacturonic acid, glucose, xylose, rhamnose and glucuronic acid gives the composition of the polymer fraction of products containing the pectin and emphasizes the natural and high level of purity of apple pectin formulated in the VitaPro ProPectin product.

Conclusions

• VitaPro ProPectin serving is equivalent to 300 g of whole apple containing 1 % of pectin or 3 kg of whole apple containing 0.1 % of pectin.

3.3.       Detoxification Properties of VitaPro ProPectin

Table 2 : Evolution of metal levels along the simulated digestion experiment with VitaPro ProPectin

sample containing 4.8 g of apple pectin.

Figure 1 : Graph view of metal level evolution along simulated digestion experiment with VitaPro ProPectin (error bars for results precision). The amount of apple pectin in the experiment was 4.8 g

(estimated from 2.2 g of Gal acid in T0).

Analytical comments

• 1 corrected from 10% mass loss induced by UF membrane fouling
• There is a very good match between results of duplicated experiments
• All metal salts are highly soluble. Without chelation they pass through the UF membrane
• The levels of metals were selected to be consistent with commonly recognized toxic levels in food. Limits for heavy metals daily intake recommended by FDA, EPA and ATSDR1 are (averages, for a body weight of 70 kg) 2:
  • 20 μg/day for Pb
  • 10 μg/day for Cd
  • 20 μg/day for Sn
  • 12 μg/day for Hg
  • VitaPro ProPectin chelates Pb, Cd, Sn and Hg and so prevent their diffusion through

Results Discussion

Conclusions

the 3 kDa UF membrane. The chelating properties range up to 700% of the maximum daily dose for toxic metals.

• Extrapolated to real human digestion, it means that VitaPro ProPectin significantly decrease Pb, Cd, Sn and Hg absorption during stomach and early intestinal phases of the digestion. So, these toxic metals are not transferred to blood circulation at this stage of digestion, preventing their potentially detrimental effects.
• These chelation data were established using 4.8 g of pectin (about 18 g of ProPectin

raw product) in contact with metal levels from 50 to 300 ppb during digestion experiment. In order to compare to other products these results need to be normalized with the level of pectin in servings (see table below).

Table 3 : VitaPro ProPectin detoxification properties for metals. Results are expressed in µg of metals chelated per serving and per day, assuming 1 servings Iday as recommended by VitaPro. Each serving

contains 3 g of pure apple pectin.

1  www.atsdr.cdc.gov

2 American Herbal Products Association. December2009. Heavy Metals: Analysis and Limits in Herba Dietary Supplements. AHP A: Silver Spring, MD

4.       Report Conclusions

4.1.       Product compositions

• In the table below we present a comparison in whole apple equivalent (assuming 0.2 % pectin level in natural apples and 150 g as average weight of apple)

• The VitaPro ProPectin is an optimized formulation containing 27 % of pure

pharmaceutical grade apple pectin and natural fructose. The profile of oses, galactose, galacturonic acid, glucose, xylose, rhamnose and glucuronic acid gives the composition of the polymer fraction of products containing the pectin and emphasizes the natural and high level of purity of pectin formulated in the VitaPro ProPectin product.

4.2.       Detoxification  properties

• The figure below shows detoxification properties of VitaPro ProPectin. One daily dose of VitaPro ProPectin can prevent the absorption of toxic metals equivalent to

several times the maximum daily intake limits recommended by EPA and FDA in food

products.

• One serving per day of VitaPro ProPectin can prevent the absorption of :
  • 7 times the maximum daily dose for lead (Pb)
  • 2 times the maximum daily dose for cadmium (Cd)
  • 6 times the maximum daily dose for tin (Sn)
  • 2 times the maximum daily dose for mercury (Hg)