in Nutraceutical Delivery Rahau Shirazi, Stephen Bloor, Kevin - PowerPoint PPT Presentation

Nano-vehicles in Nutraceutical Delivery Rahau Shirazi, Stephen Bloor, Kevin Mitchell 2016 Lipids and Nutraceuticals Conference Nelson November 2016

- Global Nutraceuticals Market - Lipophilic Vitamins – Encapsulated or Exposed? - Food grade Nano-vehicles - Encapsulated Vitamins – Preparation to Characterization - Conclusion and Future direction

Global Nutraceuticals Market Market Reports World October 28th, 2016 Analysts forecast the global Functional Food and Nutraceuticals market to grow at a CAGR (compound annual growth rate) of 7.39% during the period 2016-2020. The global dietary supplements market expected growth by 2025 - US$252 billion – from US$131 billion by 2016. Vitamin D Market – $2.5 billion by 2020 increases with growing awareness of deficiency links to health problems (e.g. osteoporosis, osteomalacia, and rickets).

Lipophilic Vitamins – Encapsulated or Exposed? Goal: Improve stability during processing and storage of fortified products. Physical and chemical factors: light (UV), heat, moisture, exposure to air, acid or alkaline environments. The biological effects of UV light: - Degradation - Safety, ex. formation of reactive species (singlet oxygen and superoxide radical anion) in case of vitamin A  Damage lipids and DNA

Food grade Nano-vehicles Key Characteristics: Food-grade protein-based and lipid- based materials, biodegradable, able to encapsulate lipophilic molecules (e.g. Retinol), easy to handle Retinol Cholecalciferol • ZSC-Formula (Vitamin A 1 ) (Vitamin D 3 ) • C-Formula + Vitamin D 3 • SSC-Formula • SS-Formula + Vitamin A 1 Encapsulated vitamin in powder form ZSC-Formula SSC-Formula SBS-Formula C-Formula Vitamin D 3 Vitamin D 3 + Vitamin A 1 Vitamin D 3

Encapsulated vitamins – Preparation to Characterization SEM – ZetaSizer Nano Zeta Potential ZSC-Formula : Vitamin D 3 pH ZSC-Formula : Vitamin D 3 500 nm 500 nm 500 nm 500 nm ZSC-Formula : 4°C to 37°C size increases from 180- ZSC-Formula + Vitamin D 3 : 4°C to 37°C size 200 nm and zeta potential changes -24 to -19 mv increases from 180-200 nm with 4.5 microm with increase in temperature particles (3%) and zeta potential: -13 at 4°C, -20 mv at 25°C, -11 mv at 37°C

Encapsulated vitamins – Preparation to Characterization SEM – ZetaSizer Nano C-Formula : Vitamin D 3 500 nm 500 nm 500 nm 500 nm C-Formula : 4°C to 37°C size increases from 160- C-Formula+Vitamin D 3 : 4°C to 37°C size slightly increases from 380-530 nm with microparticles 860 nm and zeta potential from showing(3%) at 25°C, and 80 nm particles (6%) at -10 to -13 mv with increase of temperature. 37°C and zeta potential is -25 mv at lower temperatures and -23 mv at 37°C.

Encapsulated vitamins – Preparation to Characterization SEM SSC : Vitamin D 3 Preparation: Ethanol-water, Sonication, Shear mixer, Freeze Dried 500 nm 500 nm 500 nm 500 nm Encapsulated vitamin D 3 in SSC: 4°C to 37°C size 200-300 nm. Zeta potential: -17 mv at lower temperatures and -22 mv at 37°C.

Encapsulated vitamins – Preparation to Characterization SEM SS : Vitamin A Preparation: Ethanol-water, Ultra-sonication, Freeze Dried Encapsulated 500 nm 500 nm Plant Sterols: European Food Safety Authority (EFSA) authorised claim that allows food manufacturers to state that “plant sterols contribute to the maintenance of normal blood cholesterol levels” with a “daily intake of at least 0.8g of plant sterols ” . Ref: Lecithin and Plant Sterols Nutrigold Update Service

Encapsulated vitamins – Preparation to Characterization Infant Formula Added: SS Formula + Vitamin A 1 1.00 4.00 1.20 0.80 1.15 3.00 D3 μg /mL A 1 μg /mL 0.60 D3 μg /mL 1.10 2.00 0.40 1.05 1.00 0.20 1.00 0.00 0.00 0.95 0.0 0.0 0.8 0.0 0.8 2.0 2.0 2.5 2.0 2.5 3.0 18.0 3.0 18.0 Infant Formula + Infant Formula + Infant Formula UV Exposure UV Exposure UV Exposure Vitamin D 3 Measurements Vitamin A 1 Measurements Vitamin D 3 Measurements Infant Formula Label Vitamin A 65 μ g/100 mL, Vitamin D 3 0.95 μ g/100 mL

Encapsulated vitamins – Preparation to Characterization Stability of Fortified Milk: Sun Exposure - LCMS Measurements QUARTS SHEET Fortified Milk contains vitamin D 3 6.00 5.00 4.00 D3 μg /mL ICED water 3.00 Conditions: Mixing & 2.00 Cooling in Quartz Chamber in a sunny day 1.00 0.00 0.0 4.5

Encapsulated vitamins – Preparation to Characterization Vitamin D 3 in Fortified Milk, UV - LCMS Olive-based: Vitamin D3 In Fortified Milk D3 μ g/mL 2500 2000 1500 UV UV+ 1000 Conditions: varied Volumes 500 no mixing – cooling in UV 0 chamber No UV Olive-D3 UV+ Olive-D3 UV No UV Olive Based Formula D3 UV+ Olive-Based Formula D3 UV (18 hours) Fortified Milk ZSC-Formula : Vitamin D3 D3 μ g/mL D3 μ g/mL In Fortified Milk 0.25 400 350 0.2 300 0.15 250 200 0.1 150 100 0.05 50 0 0 No UV Fortified Milk UV+ Fortified Milk UV No UV No UV ZSC-Formula + D3 UV+ Zein SC D3 UV+ Zein SC D3 UV ZSC-Formula + D3 UV (18 hours) (18 hours)

Encapsulated vitamins – Preparation to Characterization SSC : Vitamin D 3 in Fortified Milk D 3 μg /mL 30.00 UV 25.00 Sample Condition: 20.00 Mixing & Cooling 15.00 in UV Chamber 10.00 5.00 0.00 0.0 3.0 4.0 4.5 22.0 SSC:D3 in Fortified Milk - UV Exposure

Encapsulated vitamins – Preparation to Characterization Encapsulated Vitamin D 3 in Fortified Milk UV exposure – LCMS UV 4000 D 3 μg /mL Conditions: No Mixing - Cooling in UV Chamber 3000 2000 Fortified Milk + ZSC-Formula Vitamin D 3 1000 Fortified Milk + C-Formula Vitamin D 3 Fortified Milk + SSC-Formula Vitamin D 3 0 0 0.7 1.5 2 3.5 5.5 18 24 46 Time (hours)

Conclusion and Future Direction - Stability of bioactive molecules such as vitamins matters at delivery environment, storage, processing conditions. - Next we will assess effect of heat, air, acid or alkaline environments current formulas. - Future: other vitamins and bioactives in current and other encapsulation materials

Thank You The Integrated Bioactive Technologies Group (IBT) Other Presentations by Lipids Team Providing high-value solutions for the industrial biotech sector Andrew McKenzie/Dawn Scott Natural Products and Talk: Natural Sources of Plasmalogens Lipid Chemistry Plant Chemistry 10 th on 11:45am Create high-value nutraceuticals and cosmeceuticals using our lipid Proteins and bioactive Kirill Lagutin expertise peptides Poster: The use of NMR in the characterization of edible fats and oils . Fermentation & Pilot Scale Analytical Services Special Thanks to IBT Group Members Bioprocessing Facilities especially to Rosemary Webby Enzyme Technology Supercritical Fluid Extraction Funded by Strategic Investment Fund – Callaghan Innovation Process Biology and biochemistry expertise Development and application to industry