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

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.

Market Reports World

October 28th, 2016

$2.5 billion by 2020 –

increases with growing

awareness of deficiency links

to health problems (e.g.

osteoporosis, osteomalacia,

and rickets).

Vitamin D Market

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

Lipophilic Vitamins –

Encapsulated or Exposed?

Food grade Nano-vehicles

Cholecalciferol

(Vitamin D3)

Retinol

(Vitamin A1)• ZSC-Formula

• C-Formula + Vitamin D3

• SSC-Formula

• SS-Formula + Vitamin A1

Key Characteristics: Food-grade protein-based and lipid-

based materials, biodegradable, able to encapsulate

lipophilic molecules (e.g. Retinol), easy to handle

SSC-FormulaVitamin D3

C-FormulaVitamin D3

ZSC-FormulaVitamin D3

SBS-Formula + Vitamin A1

Encapsulated vitamin in powder form

ZSC-Formula : Vitamin D3

SEM – ZetaSizer Nano

ZSC-Formula: Vitamin D3

pH

Zeta

Po

ten

tial

Encapsulated vitamins – Preparation to Characterization

500 nm500 nm

ZSC-Formula: 4°C to 37°C size increases from 180-200 nm and zeta potential changes -24 to -19 mv

with increase in temperature

ZSC-Formula + Vitamin D3: 4°C to 37°C size increases from 180-200 nm with 4.5 micromparticles (3%) and zeta potential: -13 at 4°C, -20 mv at 25°C, -11 mv at 37°C

500 nm 500 nm

C-Formula : Vitamin D3

SEM – ZetaSizer Nano

Encapsulated vitamins – Preparation to Characterization

500 nm500 nm

C-Formula : 4°C to 37°C size increases from 160-860 nm and zeta potential from

-10 to -13 mv with increase of temperature.

500 nm500 nm

C-Formula+Vitamin D3 : 4°C to 37°C size slightly increases from 380-530 nm with microparticles

showing(3%) at 25°C, and 80 nm particles (6%) at 37°C and zeta potential is -25 mv at lower

temperatures and -23 mv at 37°C.

SSC : Vitamin D3

SEM

500 nm 500 nm 500 nm500 nm

Preparation: Ethanol-water, Sonication, Shear mixer, Freeze Dried

Encapsulated vitamins – Preparation to Characterization

Encapsulated vitamin D3 in SSC: 4°C to 37°C size 200-300 nm. Zeta potential: -17 mv at lower temperatures and -22 mv at 37°C.

SS : Vitamin A

SEM

500 nm500 nm

Preparation: Ethanol-water, Ultra-sonication, Freeze Dried

Encapsulated vitamins – Preparation to Characterization

Plant Sterols:European Food Safety Authority (EFSA) authorised claimthat allows food manufacturers to state that “plant sterolscontribute to the maintenance of normal bloodcholesterol levels” with a “daily intake of at least 0.8g ofplant sterols”.

Ref: Lecithin and Plant Sterols Nutrigold Update Service

Encapsulated

Infant Formula

0.95

1.00

1.05

1.10

1.15

1.20

0.02.0

3.0

D3

μg

/mL

Infant FormulaUV ExposureVitamin D3 Measurements

0.00

1.00

2.00

3.00

4.00

0.0 0.8 2.0 2.5 18.0

0.00

0.20

0.40

0.60

0.80

1.00

0.0 0.8 2.0 2.5 3.0 18.0

D3

μg

/mL

Infant Formula + UV Exposure Vitamin A1 Measurements

Infant Formula +UV ExposureVitamin D3 Measurements

A1μg

/mL

Infant Formula Label Vitamin A 65 μg/100 mL, Vitamin D3 0.95 μg/100 mL

Added: SS Formula + Vitamin A1

Encapsulated vitamins – Preparation to Characterization

Stability of Fortified Milk:Sun Exposure - LCMS Measurements

0.00

1.00

2.00

3.00

4.00

5.00

6.00

0.04.5

D3

μg

/mL

Conditions: Mixing &Cooling in QuartzChamber in a sunny day

ICED water

QUARTS SHEET

Encapsulated vitamins – Preparation to Characterization

Fortified Milk contains vitamin D3

Vitamin D3 in Fortified Milk, UV - LCMS

0

500

1000

1500

2000

2500

No UV Olive-D3 UV+ Olive-D3 UV

D3 μg/mL Olive-based: Vitamin D3 In Fortified Milk

0

50

100

150

200

250

300

350

400

No UV Zein SC D3 UV+ Zein SC D3 UV

D3 μg/mL

ZSC-Formula + D3 UV+ ZSC-Formula + D3 UV No UV

ZSC-Formula : Vitamin D3In Fortified Milk

0

0.05

0.1

0.15

0.2

0.25

No UV Fortified Milk UV+ Fortified Milk UV

D3 μg/mL Fortified Milk

UV+UV

Conditions: varied Volumesno mixing – cooling in UVchamber

(18 hours)

Encapsulated vitamins – Preparation to Characterization

No UV Olive Based Formula D3 UV+ Olive-Based Formula D3 UV

(18 hours)

(18 hours)

0.00

5.00

10.00

15.00

20.00

25.00

30.00

0.0 3.04.0

4.522.0

SSC:D3 in Fortified Milk - UV Exposure

Sample Condition:Mixing & Cooling in UV Chamber

SSC : Vitamin D3 in Fortified Milk

D3 μg/mL

UV

Encapsulated vitamins – Preparation to Characterization

0

1000

2000

3000

4000

0 0.7 1.5 2 3.5 5.5 18 24 46

Time (hours)

Fortified Milk + ZSC-Formula Vitamin D3

Fortified Milk + C-Formula Vitamin D3

Fortified Milk + SSC-Formula Vitamin D3

Encapsulated Vitamin D3 in Fortified Milk UV exposure – LCMS

D3μg

/mL

UV

Conditions: No Mixing - Cooling in UV Chamber

Encapsulated vitamins – Preparation to Characterization

Conclusion and Future Direction

- Stability of bioactive molecules such as vitaminsmatters at delivery environment, storage,processing conditions.

- Next we will assess effect of heat, air, acid oralkaline environments current formulas.

- Future: other vitamins and bioactives in currentand other encapsulation materials

The Integrated Bioactive Technologies Group (IBT)Providing high-value solutions for the industrial biotech sector

Lipid ChemistryCreate high-value nutraceuticals and

cosmeceuticals using our lipid expertise

Natural Products and Plant Chemistry

Analytical Services Pilot Scale Facilities

Process Development

Supercritical Fluid Extraction

Biology and biochemistry expertise and application to industry

Fermentation & Bioprocessing

Proteins and bioactive peptides

Enzyme Technology

Other Presentations by Lipids Team

Andrew McKenzie/Dawn ScottTalk: Natural Sources of Plasmalogens10th on 11:45am

Kirill LagutinPoster: The use of NMR in the characterization of edible fats and oils.

Special Thanks to IBT Group Members especially to Rosemary Webby

Funded by Strategic Investment Fund –Callaghan Innovation

Thank You