Predicting the release and diffusion of nisin from a polyvinyl alcohol matrix coated film. Michele Perna Kay Cooksey Duncan Darby Jeff Rhodehamel

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1 Predicting the release and diffusion of nisin from a polyvinyl alcohol matrix coated film Michele Perna Kay Cooksey Duncan Darby Jeff Rhodehamel

2 Introduction Ph.D. Sudent: Michele Perna (now at Bemis) Idea: Dissolvable antimicrobial (am) coating on package wall Selected coating basis: PVOH Selected am: Nisin

3 Ph.D. Process We like to challenge our students to perform one aspect of their dissertation that is theoretical For Michele: Since her systems depnds upon dissolution and diffusion We challenged her to consider some mass transfer implications of her work

4 Summary 1. Coating formulation 1. PVOH coating formulation containing Nisaplin 2. Effective against M. luteus 3. Potential for implementation on large scale processes 2. Large scale trials 1. Coating was able to be implemented on large scale equipment while maintaining antimicrobial efficacy against M. luteus 3. Review of diffusion and controlled release 1. Complex, multivariable system 2. Assumptions should be stated 3. Proposed methodology could be utilized for further research

5 Ph.D. Process Michele is not a chemical engineer Or a mass transfer specialist So we recommended that she consider the implications as best she could And describe a method to measure the mass transfer

6 Basic diffusion concepts Outline The antimicrobial packaging system Factors that can affect diffusion and controlled release Factors that can affect nisin efficacy Proposed methodology for future research

7 Diffusion: Basic concepts Fick s First Law (Steady State) Diffusion coefficient not dependent upon location Rate of diffusion is constant with time JJ = DD dddd dddd D = diffusion coefficient (m 2 /sec) J = mass flux (kg/m 2 *sec) C = mass per volume (kg/m 3 ) x = displacement (m)

8 Diffusion: Basic concepts Fick s Second Law (Non-steady state) Diffusion varies with time. = DD 2 CC 2 D = diffusion coefficient (m 2 /sec) C = mass per volume (kg/m 3 ) x = displacement (m) T = time (sec)

9 Diffusion: AM Packaging System y x

10 Scenarios Transfer through a solid Transfer through a gel Transfer through a liquid

11 Diffusion: Nisin Through Solid

12 Diffusion: Nisin Through Solid Pretty straight forward As long as you accept all the usual assumptions related to permeation through solids Temperature dependence does apply as this is a refreigerated application

13 Diffusion: Nisin Through Solid Low temperature application Temperature dependent Arrhenius equation DD = DD 0 eeeeee EE aa RRRR D 0 = constant (m 2 /sec) E a = Activation energy for diffusion (J/mol) R = universal gas constant (J/mol*K) T = temperature (K)

14 Diffusion: Nisin Through Gel PVOH is a highly swellable, water soluble polymer Gels upon solvent penetration Plasticization / Polymer relaxation

15 Diffusion: Nisin Through Gel

16 Diffusion: Nisin Through Liquid Assumption: Nisin would be held in the polymer matrix until release via dissolution processes.

17 Diffusion: Nisin Through Liquid

18 Diffusion: Nisin Through Liquid Release based upon coating dissolution If convection is significant: dddddddddddddddddddddd rrrrrrrr δδδδ δδδδ = h(cc LL.tt CC eeee ) D = diffusion coefficient (m 2 /sec) C = mass per volume (kg/m 3 ) x = displacement (m) h = coefficient of transfer by convection C L,t = concentration of the diffusion substance on the surface of the solid C eq = concentration of the diffusing substance on the surface required to maintain equilibrium

19 Diffusion: Nisin Through Liquid Release based upon coating dissolution If convection is not significant driving force is diffusion: dddddddddddddddddddddd rrrrrrrr δδδδ δδδδ = DD dddd dddd D = diffusion coefficient (m 2 /sec) C = mass per volume (kg/m 3 ) x = displacement (m)

20 Antimicrobial Packaging system In the non-modeling reality the nisin release or diffusion is likely to be a combination or the solid, gel and liquid scenarios.

21 Diffusion: Factors that can affect diffusion and controlled release Intrinsic factors (ph, fat content, food structure and polymer matrix,composition) Polymer swellability and structure Temperature Permeant size and distribution Food product AM concentration in the pkg/pkg structure

22 Diffusion: Factors that can affect diffusion and controlled release Rate of consumption by microorganisms Direction of flux AM solubility in the pkg system Dissolution Liquid volume Area and thickness of pkg material Convection

23 Nisin Concentration: Factors that can affect nisin efficacy ph Target microorganisms Food matrix Package production Fat content Jung, Bodyfelt and Daeschal half and half (12.9% fat) Bhatti, Veeramachaneni and Shelef 2004 milk (2-3.5% fat)

24 Diffusion: Proposed Methodology Assumptions The direction of flux for the antimicrobial is mono-directional in the direction of the food product or away from the packaging substrate Driving force = rate of consumption by microorganisms

25 Diffusion: Proposed Methodology Assumptions Packaging system release of nisin occurs via diffusion and/or convection (dependent on further testing) Mathematical modeling may require separate models for these two different modes

26 Diffusion: Proposed Methodology Assumptions If coating dissolves assumption of a moving boundary condition. As coating dissolves, nisin concentration may remain constant but its location in the system will change. If the coating gels there is no moving boundary as in the item above.

27 Diffusion: Proposed Methodology Assumptions The nisin is mixed homogeneously throughout the coating. No nisin remains trapped within the coating matrix once the matrix is completely dissolved. The concentration of nisin at the coating-liquid interface is equal to the initial concentration as the release occurs. (C t,0 = C 0 )

28 Diffusion: Proposed Methodology Film on lawn with standard curve Test different microbes spoilage RTE Lactobacillus spp. Lueconostoc spp. Serratia spp. Brochothrix thermosphacta Enterococcus casseliflavus Zone of Inhibition Treatment Film Control Film

29 Diffusion: Proposed Methodology Dissolution Large # of samples Remove at certain periods, blot to dry, coat weight Plot coat weight over time Food simulant water or hotdog brine

30 Diffusion: Proposed Methodology Nisin Quantification BCA Bicinchoninic Acid protein assay Spectrophotometric Method Measure absorbance at 562 nm Purple color Cu(II) to Cu(I); alkaline conditions

31 Diffusion: Conclusions Highly swellable and water soluble polymers Can be used for controlled release systems in active packaging applications Various factors Affect both diffusion and nisin efficacy

32 Diffusion: Conclusions There is much work to be done to better understand controlled release systems in addition to the packaging system proposed.

33 Future research recommendations 1. Determine the sensitivity of other spoilage microorganisms against film produced. 2. Additional material properties such as heat seal curves and thermoforming capabilities. 3. Conducting a challenge study on an actual food product 4. Diffusion and release studies are recommended to better understand the packaging system

34 Thank you!!!

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