Properties of spray-dried food flavours microencapsulated with twolayered membranes: Roles of interfacial interactions and water

Adem Gharsallaoui, Galle Roudaut, Laurent Beney, Odile Chambin, Andre Voilley, Rmi Saurel.

GUIDE:- Prof. R. S. Singhal

Presented by Mahesh Kharat M.Tech FET (I)

Introduction
Microencapsulation is defined as a process in which tiny particles or droplets are surrounded by a coating, or embedded in a homogeneous or heterogeneous matrix, to give small capsules with many useful properties

The material inside the microcapsule is referred to as the core, internal phase or fill, whereas the wall is sometimes called shell, coating, wall material, or membrane. Wall material should posses various properties so as to ensure long term protection of core ingredient.

Introduction
Coating material properties: Stabilization of core material Inert toward active ingredients Controlled release under specific conditions Film-forming, pliable, tasteless, and stable Non-hygroscopic, less viscosity, economical The coating can be flexible, brittle, hard, thin etc. Coating materials: Gums: Gum arabic, sodium alginate, carageenan. Carbohydrates: Starch, dextran, sucrose Celluloses: Carboxymethylcellulose, methycellulose. Lipids: Bees wax, stearic acid, phospholipids. Proteins: Gelatin, albumin.

Introduction
Shahidi and Han (1993) proposed six reasons for applying microencapsulation in food industry:
1. To reduce the core reactivity with environmental factors. 2. To decrease the transfer rate of the core material to the outside environment.

3. To promote easier handling.

4. To control the release of the core material.

5. To mask the core taste.

Introduction
Depending upon number of cores present, and wall materials used to protect the same there exist different types of morphologies of microcapsules e.g. simple, multi-core, multi-wall etc. (Gibbs et al. 1999).

Introduction
Amongst all physical and chemical ways, spray drying provides a promising way for microencapsulation, particularly in food industry, basically due to its low cost and god product quality.
Ingredients flavours Components
L-Menthol D-Limonene

reference Soottitantawat et al., 2005b Soottitantawat et al., 2005a, 2004 Krishnan et al., 2005 Kanakdande et al., 2007 Ribeiro et al., 2003 Santipanichwong & Suphantharika, 2007 Ribeiro et al., 2005

lipids and oleoresins

cardamom cumin

Other functional components

lycopene -carotein

astaxanthin

Introduction
Multilayer emulsions and multilayered microcapsules thus obtained after spray drying are thought of as an effective technique these days. Some of their advantages are (Guzey & McClements, 2006): 1. Emulsion stability improvement; 2. Increased chemical stability of encapsulated compounds; 3. Controlled release rate of functional molecules 4. Ability to set off the encapsulated functional agent release by changing the environmental conditions such as dilution, pH, ionic strength, or temperature.

Introduction

Summary of the paper

Aim:

To determine whether the multilayer coatings can be used to improve flavor protective properties of pea protein-coated oil droplets.

Summary of the paper

Methodology : Three flavour compounds viz. ethyl acetate, ethyl butyrate, & ethyl hexanoate belonging to strawberry flavour note, were chosen. Mono & multi-layer emulsions were prepared using following:
1. 2. 3. 4. Preparation of PPI solution (aqueous). Preparation of three different miglyol (10%) solutions containing flavor components (non-aqueous). Triglyceride solution blended with 90% wt. aqueous solution (17000 rpm, for 90 sec). Imidazol/acetate buffer (5mM, pH 2.4)or pectin solution (prepared in the same buffer) were added to obtain PRIMARY or SECONDARY emulsions respectively.

Summary of the paper

Methodology : Spray drying of emulsions was carried out to get encapsulated flavor powders, under following conditions: Feed rate :1 L/h 180 0C 90 0C 0.5mm

Inlet temperature : outlet temperature : nozzle atomizer :-

 Properties of encapsulated flavors studied and the methods used are described below:1. Study of flavor retension in dried powder - reconstitution of powders with buffer. - hexane extraction of flavors. - analysis by gas chromatography - 5-nonanol was used as standard 2. Fourier-transform infrared spectroscopy (FTIR) of proteinpectin complexes (800 4000 cm-1)

Summary of the paper

3. Release of flavors from spray dried powders - placing both freshly prepared emulsions and reconstituted in various constant humidity environments. - quantification of residual flavors by GC, after hexane extraction.
4. Differential scanning calorimetry (DSC) analysis done between -100 0C to +160 0C with empty aluminium pan as reference.

Observations and Results

1. Flavor retention in liquid emulsions a. Flavor retention increases as chain length increases.
b. No significant difference between retention in primary & secondary emulsions. The reason, according to author, for this is that the pectin is added in last stage of preparation.

c.

Observations and Results

2. Effect of pectin on flavor retention a. Increased retention with increase in chain length. b. All secondary powders showed same retention ( 65%) better than all primary powders. c. Claims :1.semipermeable matrix formation as according to Selective Diffusion Theory (Thijssen & Rulkens, 1968), during initial stages of drying. 2.Protective effect of pectin against heat induced loss of the -sheet structure in pea proteins.

Observations and Results

3. Effect of pectin on pea protein thermal sensitivity
a. Protein-pectin complexation does not prevent general molecular reorganization of pea globulins upon heating.

b.

However, it has a strong protective effect against heat induced destability of -sheet structure.

Observations and Results

4. Release properties of encapsulated flavors from spray dried powders:
a. At low relative humidity, release of flavor from spray dried powder was greater than that of emulsions. Lowest flavor release rate was observed in the range 33-52% RH.

b.

Conclusion
Inference : The interfacial membrane properties seemed to play an important role in increasing volatile compounds retention during spray-drying. Flavor diffusion could be linked to the thermal denaturation of adsorbed pea protein leading to molecular changes that can lead to modification of the permeability of interfacial layers to flavor compounds. Conclusion: Two-layered emulsions use for microencapsulation, increases flavor retention in final dry powders. Pectin inhibits the loss of -sheet protein structure at increased temperatures. This effect in addition to thicker interfacial two-layered membrane explains the above claim made.

Significance of area of research

The addition of maltodextrin to the pea protein wall material increases the particle size, (Pierucci et al., 2006). However, addition of pectin (0.2%) results in thicker walls, hence less steric repulsion Protection of -sheet protein structure. Polysaccharide/protein interactions give new functions to pea proteins without chemical or enzymatic modification, particularly solubility, foaming and surfactant properties (Liu et al., 2010). These interactions gives, better particle size distribution Stable emulsions (creaming stability) Improved efficiency of the microencapsulation.

Literature Survey
McClements et al., (2005) carried out studies in which tuna oil-inwater emulsions containing droplets stabilized by lecithinchitosan membranes. Layer-by-layer electrostatic deposition method was used that involved adsorption of cationic chitosan onto the surface of anionic lecithinstabilized droplets.
The secondary emulsions showed better stability to droplet aggregation than primary emulsions exposed to
thermal processing (3090 0C for 30 min) freeze-thaw cycling (-18 0C for 22 h/30 0C for 2 h) high sodium chloride contents (200 mM NaCl) and freeze-drying.

Literature Survey

Studies mentioned above were further continued by McClements et al., (2006) in which Corn syrup solids were added to the emulsions.
Spray-dried powders ,

had relatively low moisture contents (<3%), high oil retention levels (>85%) and rapid water dispersibility (<1 min) The structure of the microcapsules was unaffected by drying temperature from 165 to 195 0C.

Literature Survey
Pierucci et al. (2007) -tocopherol were produced by spray drying technique using pea protein (PP), carboxymethylcellulose(CMC) and mixtures of these materials with maltodextrin (PP-M and CMC-M) as wall materials.

The results showed that the pea protein use for -tocopherol microencapsulation is a promising system for further application in food.

The retention of -tocopherol was above 77%, mean particles size remained below 7 m.

Literature Survey
Gharsallaoui et al., (2010a) studied the effect of high methoxyl pectin on pea protein isolates in aqueous solution and at oil/water interface.
Addition of pectin improved emulsion stability to pH changes and depletion flocculation induced by maltodextrin addition. This could be mainly assigned to steric repulsion and oilwater interfacial membrane rigidity improvement after pectin adsorption.

Literature Survey
Gharsallaoui et al., (2010b) extended above studies to spray dried emulsion.
Emulsions showed better stability to droplet aggregation after drying as compared to those containing only pea protein as wall material. Pectin, an anionic polysaccharide, forms a less charged protective layer around the protein interfacial film surrounding the oil droplets that improved their stability to spray-drying mainly by increasing steric effects.

Critical Analysis
Important features of paper Pea protein pectin : A technically correct combination

o Protein gets completely dissociated at acidic pH s. o Anionic pectin regions perfect to be electro-statically deposited over protein (containing NH2 groups). o Pectin-a functional ingredient in food . o Protein-highly nutritional values.

Significant correlation is found between use of pectin as wall material and retention (and stability) of volatile ester flavors.
All above mentioned studies were done using high precision methods with standard protocol.

Originality
Layer-by-layer (LBL) deposition system in formation and studying of multilayered emulsions have already been used by various authors.

Authors
Faldt et al. (1993) Magdassi et al. (1997) Guzey et al. (2004) Ogawa .S, Decker E. A., McClements D. J. (2004)

Multilayer wall system

phosphatidyl cholinechitosan chitosanlecithin

lecithinchitosan pectin

-lactoglobulinpectin

Originality
Some studies on multi-layered microencapsulations have already been done and successfully reported. For example, 1. McClements et al. (2006) studied about lipid oxidation, free oil & encapsulation efficiency, powder color & morphology, etc. of spray dried tuna oil microencapsulated with lecithin-chitosan membranes. Also, pea proteins are many a times used along with polysaccharides in several microencapsulation studies.

Wall material
Protein Pea globulins Polysaccharide Gum Arabic, CMC, sodium alginate

Core material
Triglyceride

Reference

Ducel et al. (2004a,b)

Pea proteins
Pea proteins Pea proteins

Maltodextrin
Maltodextrin Maltodextrin

Ascorbic acid
-Tocopherol Ascorbic acid

Pierucci et al. (2006)

Pierucci et al. (2007) Pereira et al. (2009)

Originality
The idea is not novel. However, this combination of wall materials were not studied before. Very similar studies have already been carried out by the same author (Gharsallaoui et al., 2010a,b) where the aim was to study the stability parameters; this manuscript rather emphasizes on understanding of interaction chemistry. The latter is well thought, and explained with support of observations as well as logical assumptions. Length of the paper is acceptable, though introduction part was found to be too lengthy.

Technical correctness
Technically correct
o Standard methods used. o Analysis done in triplicates. o Results were expressed as mean standard deviation(SD).

Assumptions were reasonable Reasoning was logical

Technical correctness
However, following point must have been considered: 1.introduction, para-3, lines:1 o Argument made about past studies by McClements et al. (2005) is wrong, where tuna oil studies are mentioned as -3 fatty acid studies .

2.2 Preparation of mono and multilayered emulsions containing flavors o 0.555 wt.% PPI solution was used to prepare final 10 & 20 emulsions containing 0.25 wt.% PPI. o Filtration loss of protein is not taken under consideration. o If, in case considered, it has not been mentioned.

Technical correctness
3.1. Flavor retention in liquid emulsions
o No significant diffrence in ethyl ester retention between primary & secondary emulsion, as pectin was added in last stages of preparation.

o This does not appears logical as stability is determined after keeping the emulsion without disturbance for a long time.

Technical correctness
3.4 effect of heating on pea protein amide 1 band o Author says that There was general decrease in the intensity of -turns band (1669 cm-1) . o This is not presented in the respective graph.

Clarity
Paper is complete with the work as it should be according to topic stated. Paper always stuck to its subject. Language easy to follow and understand.

Bibliography
The author cites all the references in the text and vice versa.
The manuscript accurately represents statements in cited references. All the references are complete and as per guideline except those mentioned in following table:

Bibliography
Reference Dong et al., 1990 Fang Y., Dalgleish D. G.,1997 Mistakes/corrections Page no's are (3303-3308), not (347-354) Reference given: Journal Dairy Science, 50, 1363-67 correct reference: Journal of colloid interface science, 196(2), 292-298 Volume 24 is written, issue 3 is missing Volume 88 is written, issue 1 is missing Volume 38 is written, issue 5 is missing Volume 52 is written, issue 5 is missing Exact reference was not found

Grigoriev et al., 2008 Hopp T. P., 1986 Rosenberg et al., 1990 Soottitantawat et al., 2004 Thijssen, H. A. C., & Rulkens, W. H. (1968).

Title and Abstract

Title is suitable for the paper and it is self explanatory. Title is Properties of spray-dried food flavours microencapsulated with two-layered membranes: Roles of interfacial interactions and water

However, only esters are studied; hence whether these studies of flavor retention in powders, release characteristics could be extrapolated to various flavor components e.g. polyphenols, aldehydes, ketones, terpenoids, etc. is still questionable.
Abstract is not too wordy, & brings out the main points of the paper.

Illustrations and Tables

Illustrations are clear and legible. All the results are well explained with graphical representations wherever required. All observations plotted on graphs were represented with standard error bars. However, in 3.6 Release properties of encapsulated flavours from spray dried powders: graphs are labelled as HR 33%, HR 52 % & so on instead of RH 33%, RH 52% etc. for relative humidity.

Suggestions
Emulsion stability study including parameters such as particle size distribution, zeta-potential could have been done.

The release properties of encapsulated flavors has been studied only with respect to one parameter i.e. in various environments with particular relative humidity. Other parameters such as temperature, pH, ionic strength could have been included.
t1/2 values studies could have been done while studying the stability of the encapsulated powders, which will give the exact time required to degrade 50% of the initial flavor concentration.

Suggestions
Comparative studies with the reconstituted emulsions are also significant, but was not carried out.

Morphological studies, which are not done, would have made this manuscript more complete.

Suggestions
Studies with several flavoring components needed to draw any generalized conclusion.

Transglutaminase treatment of pea proteins to further modify their functional properties, may be thought of (Shand P. J. et al., 2008).
Fairly harsh conditions during emulsion preparation (mixing, blending (17500 rpm) & homogenization @ 500 bar). A lot of flavor loss takes place here only. Repeated membrane emulsification could be one alternative (Ribeiro et al., 2005). This should be included in similar future work.

References
Ducel, V., Richard, J., Popineau, Y., Boury, F., 2004a. Adsorption kinetics and rheological interfacial properties of plant proteins at the oilwater interface. Biomacromolecules, 5, 20882093. Ducel, V., Richard, J., Popineau, Y., Boury, F., 2005. Rheological interfacial properties of plant protein-Arabic gum coacervates at the oilwater interface. Biomacromolecules, 6, 790 796.

Gharsallaoui, A., Saurel, R., Chambin, O., Cases, E., Voilley, A., & Cayot, P. (2010a). Utilisation of pectin coating to enhance spray-dry stability of pea proteinstabilised oil-in-water emulsions. Food Chemistry, 122(2), 447454. Gharsallaoui, A., Yamauchi, K., Chambin, O., Cases, E., Saurel, R., (2010b). Effect of high methoxyl pectin on pea protein in aqueous solution and at oil/water interface. Carbohydrate Polymers, 80, 817-827.

Faldt P, Bergenstahl B, Claesson PM., (1993). Stabilization by chitosan of soybean oilemulsions coated with phospholipid and glycocholic acid. Colloids Surf A Physicochem Eng Asp, 71(2), 18795.

References
Gibbs, B. F., Kermasha, S., Alli, I., & Mulligan, C. N. (1999). Encapsulation in the food industry: A review. Internatioanl Journal of Food Sciences and Nutrition, 50, 213224. Guzey, D., & McClements, D. J. (2006). Formation, stability and properties of multilayer emulsions for application in the food industry. Advances in Colloid and Interface Science, 128130, 227248.

Guzey D, Kim HJ, McClements DJ., 2004. Factors influencing the production of o/w emulsions stabilized by beta-lactoglobulinpectin membranes. Food Hydrocolloids, 18(6),96775. Kanakdande, D., Bhosale, R., & Singhal, R. S. (2007). Stability of cumin oleoresin microencapsulated in different combination of gum arabic, maltodextrin and modified starch. Carbohydrate Polymers, 67, 536541.

Liu, S., Elmer, C., Low, N.H., Nickerson, M.T., 2010. Effect of pH on the functional behaviour of pea protein isolate-gum Arabic complexes. Food Research International, 43, 489495.

Krishnan, S., Bhosale, R., & Singhal, R. S. (2005). Microencapsulation of cardamom oleoresin: Evaluation of blends of gum arabic, maltodextrin and a modified starch as wall materials. Carbohydrate Polymers, 61, 95102.

References
Magdassi S, Bach U, Mumcuoglu KY., 1997. Formation of positively charged microcapsules based on chitosanlecithin interactions. Journal of Microencapsulation, 14(2), 18995. Ogawa S, Decker EA, McClements DJ., 2004. Production and characterization of O/W emulsions containing droplets stabilized by lecithinchitosan pectin mutilayered membranes. Journal of Agriculture & Food Chemistry, 52(11), 3595600.

Pereira, H.V.R., Saraiva, K.P., Carvalho, L.M.J., Andrade, L.R., Pedrosa, C., Pierucci, A.P.T.R., 2009. Legumes seeds protein isolates in the production of ascorbic acid microparticles. Food Res. Int., 42, 115121.

Pierucci, A.P.T.R., Andrade, L.R., Baptista, E.B., Volpato, N.M., Rocha-Leao, M.H.M., 2006. New microencapsulation system for ascorbic acid using pea protein concentrate as coat protector. Journal of Microencapsulation, 23, 654662.
Pierucci, A.P.T.R., Andrade, L.R., Farina, M., Pedrosa, C., Rocha-Leao, M.H.M., 2007. Comparison of a-tocopherol microparticles produced with different wall materials: pea protein a new interesting alternative. Journal of Microencapsulation, 24, 201213.

Ribeiro, H. S., Ax, K., & Schubert, H. (2003). Stability of lycopene emulsions in food systems. Journal of Food Science, 68(9), 27302734.

References
Magdassi S, Bach U, Mumcuoglu KY., 1997. Formation of positively charged microcapsules based on chitosanlecithin interactions. Journal of Microencapsulation, 14(2), 18995.

Ogawa S, Decker EA, McClements DJ., 2004. Production and characterization of O/W emulsions containing droplets stabilized by lecithinchitosan pectin mutilayered membranes. Journal of Agriculture & Food Chemistry, 52(11), 3595600.

Pereira, H.V.R., Saraiva, K.P., Carvalho, L.M.J., Andrade, L.R., Pedrosa, C., Pierucci, A.P.T.R., 2009. Legumes seeds protein isolates in the production of ascorbic acid microparticles. Food Res. Int., 42, 115121. Pierucci, A.P.T.R., Andrade, L.R., Baptista, E.B., Volpato, N.M., Rocha-Leao, M.H.M., 2006. New microencapsulation system for ascorbic acid using pea protein concentrate as coat protector. Journal of Microencapsulation, 23, 654662.

Pierucci, A.P.T.R., Andrade, L.R., Farina, M., Pedrosa, C., Rocha-Leao, M.H.M., 2007. Comparison of a-tocopherol microparticles produced with different wall materials: pea protein a new interesting alternative. Journal of Microencapsulation, 24, 201213. Ribeiro, H. S., Ax, K., & Schubert, H. (2003). Stability of lycopene emulsions in food systems. Journal of Food Science, 68(9), 27302734.

References
Soottitantawat, A., Bigeard, F., Yoshii, H., Furuta, T., Ohkawara, M., & Linko, P. (2005a). Influence of emulsion and powder size on the stability of encapsulated D-limonene by spray drying. Innovative Food Science and Emerging Technologies, 6, 107114.

Soottitantawat, A., Yoshi, H., Furuta, T., Ohgawara, M., Forssell, P., & Partanen, R. (2004). Effect of water activity on the release characteristics and oxidative stability of D-limonene encapsulated by spray drying. Journal of Agriculture and Food Chemistry, 52, 12691276.