VITAPRO. Detoxification properties of Apple pectin formulation, PROPECTIN. CENTRE D ETUDE ET DE VALORISATION DES ALGUES FINAL REPORT.

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1 CENTRE D ETUDE & DE VALORISATION DES ALGUES 2400 Detxificatin prperties f Apple pectin frmulatin, PROPECTIN. VITAPRO FINAL REPORT PrPectin July 2015 CENTRE D ETUDE ET DE VALORISATION DES ALGUES PRESQU ILE DE PEN-LAN - B.P.3 F PLEUBIAN TEL : +33 (0) FAX : +33 (0) ALGUE@CEVA.FR --

2 1. Recall VitaPr is a cmpany selling a patented frmulatin f pharmaceutical grade apple pectin as a nutritinal supplement fr detxificatin, chlesterl management and bld glucse management applicatins. Centre d Etude et de Valrisatin des Algues (CEVA) was established in It prvides t its custmers expertise, infrmatin centre, labratries and pilt plant facilities fr studies, and develpment f innvating prducts. 2. Wrk prgramme 2.1. Biavailability definitin and expertize at CEVA Biavailability is defined as the amunt f nutrient digested, absrbed and metablized by the rganism. It can als be defined as the amunt f nutrient, after absrptin, used in bilgical prcesses. There are tw majr cmpnents f biavailability : the absrptin, which is the nutrient transfer thrugh intestine cell wall and the use f the nutrient in metablism. Then a nutrient having a fast diffusin, but being quickly released by the rganism, will get a lw biavailability. There is a significant market trend tward healthy fd and fd supplements. Infrmatin abut chemical cmpsitin f fd is mre and mre available t cnsumers, even fr lw cncentratin nutrients (minerals, vitamins, ) hwever there are nly seldm infrmatin n biavailability. Fr instance, fd rich in irn culd bring little irn t rganism if it cntains chelating agents fr irn, preventing it t be absrbed during digestin. On the ther hand, sme cmpunds like ascrbic acid can stimulate irn uptake. The cnsumer cannt manage t knw all this infrmatin. The supplier, based n data generated by apprpriate analysis and cmpared t ther prducts, shuld bring it t the cnsumer. CEVA participated in a prject t develp biavailability understanding fr Calcium and Idine in varius fd and ingredient. This allwed t install new analytical testing and t build a database f biavailability data. The testing is based n digestin mimicking, with several steps f acid and basic digestin including use f digestin enzymes. UF membranes are used t mimic intestine cell wall. VITAPRO - PROPECTIN PAGE 2/11

3 2.2 Prpsed study, extract Detxificatin Prperties f apple pectin The aim f this part is t quantify the chelating effect f PROPECTIN n selected heavy metals, knwn as txic fr human health. A knw n amunt f apple pectin extract will be set in chemical cnditins mimicking human digestin (see belw). A mixture f heavy metals (Cd, Sn, Hg, Pb) will be added t the slutin at cncentratins abve the average security limit defined fr daily fd intake. Samples will be analysed fr individual metal cncentratins using ICP-OES techniques. Sampling will be perfrmed alng the digestin prcess, as described abve. The experiment will run at the same time as Part 1, using same samples. In case f high detx prperties, metals will be chelated by pectin and will nt pass thrugh the UF membrane. In case f weak detx prperties, metals will pass the membrane. VITAPRO - PROPECTIN PAGE 3/11

4 3. Results 3.1. Sampling Strategy Initial raw prduct tested in ur experiments : Our References 2015-NV-003 Custmer References VITAPRO PROPECTIN (pwder) Step 1 : Slutins befre digestin experiments Slutins f pectin samples disslved in water = T 0 Step 2 : Gastric digestin (ph 2, 37 C, with pepsin) After 1 H at 37 C : sampling f the slutin = T 1 Step 3 : Partial intestinal digestin (ph 7.5) 2 H at 37 C, then 1 H at 4 C t stp prcess Centrifugatin f the slutin, sampling f supernatant = T 2 Sampling f slid phase (if any) = cult Step 4 : Simulatin f intestinal absrptin Filtratin trugh Ultra-Filtratin membrane set at Da (3 kda). Retentat = T 3 Permeat = T 4 VITAPRO - PROPECTIN PAGE 4/11

5 3.2. Part 1 Analysis f PrPectin Our References Custmer References 2015-NV-003 VITAPRO PROPECTIN Precisins f the analysis Entire Prduct Dry matter % Mineral cntent % Disslved Plysaccharides Galactse % Glucse % Xylse < % Rhamnse % Galacturnic acid % Glucurnic acid < % Mlecular weight % Table 1 : Analytical results PrPectin. All cncentratins are expressed in %/raw = % in riginal prduct as it is delivered. Results discussin Natural Apple pectins are made f a mixture f plysaccharides rich in Galacturnic acid (Gal acid). Cmpsitin may vary accrding t apple species, seasn and extractin prcedures, hwever a pure natural apple pectin is always rich in Galacturnic acid. Cnclusins The VITAPRO PROPECTIN is an ptimized frmulatin cntaining 27 % f pure pharmaceutical grade apple pectin and natural fructse. The prfile f ses, galactse, galacturnic acid, glucse, xylse, rhamnse and glucurnic acid gives the cmpsitin f the plymer fractin f prducts cntaining the pectin and emphasizes the natural and high level f purity f pectin frmulated in VITAPRO PROPECTIN prduct. VITAPRO - PROPECTIN PAGE 5/11

6 VITAPRO PROPECTIN serving is equivalent t 300 g f whle apple cntaining 1 % f pectin r 3 kg f whle apple cntaining 0.1 % f pectin.. VITAPRO - PROPECTIN PAGE 6/11

7 RESULTS VITAPRO - PROPECTIN PAGE 7/11

8 3.4. Part 2 - Detxificatin Prperties f apple pectin extract NV VITAPRO PROPECTIN Our References Expected NV-043 NV-048 NV-044 NV-049 NV-045 NV-050 NV-046 NV-051 NV-047 NV-052 Sample Descriptin T x T 0 befre digestin T 1 after digestin T 2 supernatant T 3 retentat 1 T 4 permeat Ttal Metals Lead, Pb (μg) Cadmium, Cd (μg) Tin, Sn (μg) Mercury, Hg (μg) Table 3 : Evlutin f metal levels alng the simulated digestin experiment with VITAPRO PROPECTIN sample cntaining 4.8 g f pectin. Figure 2 : Graph view f metal level evlutin alng simulated digestin experiment with VITAPRO PROPECTIN (errr bars fr results precisin). The amunt f pectin in the experiment was 4.8 g (estimated frm 2.2 g f Gal acid in T 0). VITAPRO - PROPECTIN PAGE 8/11

9 Analytical cmments 1 crrected frm 10% mass lss induced by UF membrane fuling There is a very gd match between results f duplicated experiments Results Discussin All metal salts are highly sluble. Withut chelatin they pass thrugh the UF membrane The level f metals were selected t be cnsistent with cmmnly recgnized txic levels in fd 300 ppm (μg/l) fr Pb 50 ppm (μg/l) fr Cd 250 ppm (μg/l) fr Sn 50 ppm (μg/l) fr Hg Cnclusins VITAPRO PROPECTIN chelates Pb, Cd, Sn and Hg and s prevent their diffusin thrugh the 3 kda UF membrane. The retentin rate ranges frm 98 t 100 %. Extraplated t real human digestin, it means that VITAPRO PROPECTIN significantly decrease Pb, Cd, Sn and Hg absrptin during stmach and early intestinal phases f the digestin. S, these txic metals are nt transferred t bld circulatin at this stage f digestin, preventing their ptentially detrimental effects. These chelatin data were established using 4.8 g f pectin (abut 18 g f PROPECTIN raw prduct) in cntact with metal levels frm 50 t 300 ppm during digestin experiment. In rder t cmpare t ther prducts these results need t be nrmalized with the level f pectin in servings (see table belw). Chelatin Prperties μg / serving μg / day Lead, Pb Cadmium, Cd Tin, Sn Mercury, Hg Table 4 : VITAPRO PROPECTIN detxificatin prperties fr metals. Results are expressed in μg f metals chelated per serving and per day, assuming 1 servings /day as recmmended by VITAPRO. Each serving cntains 3 g f pure pectin. VITAPRO - PROPECTIN PAGE 9/11