SPEAKER NOTES

Slide 1: Title Page

Slide 2: Screenshot of reports front page

Slide 3: Introduction

Notes: Long chain ω-3 PUFAs are an important nutrient with health benefits such as

antithrombotic, antiatherosclerotic, anti-inflammatory, and antiarrhythmic effects. Emulsion

substance, form, and shelf life are thought to play a major role in determining emulsion stability.

Natural antioxidants have become increasingly popular due to client preferences.

Slide 4: Aim and Objective

Notes: In this study, cod skin collagen peptides (CSCP) and γ-carrageenan (γ-car) were

conjugated using the Maillard process to create antioxidant emulsifiers. The γ-car and CSCP

mixes were first cooked under regulated Maillard reactions in a dry condition for this purpose.

Finally, by measuring the peroxide levels in the model system, The conjugate-stabilized

emulsions' oxidative stability was assessed.

Slide 5: Materials and Methods

Notes: Following a two-hour period of stirring the mixture while suspending CSCP and ι-

car (1:4, m/m) in phosphate buffer (pH 7.0, 0.05 mmol/L)., freeze-drying was performed to

create a homogeneous dispersion. The mixtures were dried, then they were grown in a

desiccator with a supersaturated KBr solution (79% relative humidity) at 60 °C. Samples
were taken from each combination after 0-2-4-6-8-10-12-14 days, and they were kept at

−20 °C until being used.

Slide 6: Molecular Weight Distribution

Notes: Using gel-permeation chromatography and an Xtimate SEC-120 column (300 × 7.8 mm,

5 μm) based on an Elite P230 high-performance liquid chromatography (HPLC) system, the

molecular weight (MW) distribution profiles of peroxides were determined. Samples were

filtered via a 0.22 μm filter to create 1 mg/mL solutions, which were then injected onto the

HPLC column in 40 µL increments.

Slide 7: Determination of the Degree of the Graft

Notes: A few little changes were performed in order to analyze free amino groups and indirectly

measure the degree of graft (DG). The solution was diluted with water until a volume of 50

milliliters was obtained. In summary, 4 mL of OPA and 200 μL of sample solution were

combined in the tube, and the mixture was let to react for 2 minutes at 35 °C in a water bath.

Slide 8: Determination of Emulsifying Capacity

Notes: Following the addition of homogenisation at room temperature for one minute at 20,000

rpm using a T25 Easy clean disperser (IKA, Staufen, Germany) after the diluted sample was

added to the AKO at a volume ratio of 5:95.

The emulsifying activity index (EAI) and emulsifying stability index (ESI) were calculated using

the equations on the slide.

Slide 9: Antioxidant Activity

Notes: To evaluate the DPPH radical scavenging activity (RSA), this methodology was

modified. Following that, the samples were incubated in the dark for 30 minutes, during which

time the absorbance at 517 nm was measured. Every sample was evaluated three times. The

equation on the slide was used to determine the samples' DPPH RSA.

Slide 10: Statistical Analysis

Notes: To get mean results, each study was conducted in triplicate. The data are then

summarized as mean ± standard deviation (SD). One-way analysis of variance (ANOVA) was

used to analyze the significance between groups, and p < 0.05 was used to indicate the

significant difference analysis of the data.

Slide 11: Results and Discussions

Notes: Molecular Composition of Peptides:

First Figure in the slide displays the findings of the peptide

distribution. According to the results, the peptides with a molecular

weight of less than 1000 Da (74.5%) were more prevalent in the CSCP

sample than those with a molecular weight of more than 1000 Da

(25.5%). It was discovered that the mean CSCP MW was 2146 Da. The FTIR

spectra of the CSCP are shown in the Figure. The amide I at 1652 cm−1

and amide II at 1538 cm−1 were the CSCP peaks that indicated the

stretching of the Cdouble bondO and Nsingle bondH, respectively.

Slide 12: Results and Discussions

Notes: Characteristics of Maillard reaction products:

With FT-IR, the secondary structure of proteins or polypeptides is regularly analyzed in

spectroscopy. It mainly examines the peptide chain structural alterations by means of the

usual distinct peaks that appear in the mid-infrared range. The absorption peak of the γ-car-

CSCP MRPs sugar conjugate was larger in the native CSCP than in the 3000–3500 cm−1

range because of the hydrogen bond stretching vibration or the single bond's bending

vibration.

Slide 13: Results and Discussions

Notes: Maillard reaction's impact on emulsion preparation:

Glycosylated proteins' capacity to emulsify was assessed using ESI and EAI. The slide's figure

displays the γ-car-CSCP MRPs' EAI and ESI at different reaction times. The conjugates' EAI and

ESI were, as predicted, significantly higher than those of γ-car-CSCP because ι-car was

covalently bound to a protein to create a macromolecular stabilizing layer surrounding oil

droplets and stabilize them against creaming, coalescence, and flocculation.

Slide 14: Results and Discussions

Notes: The stabilized emulsions' FFA release and quercetin's bioaccessibility:

FFAs from the MRPs-AKO emulsion were released at all stages of the digestion process,

suggesting that AST was released into the system. FFAs produced by the emulsions during small

intestine digestion are shown in the first figure.

After digestion, the amount of AST that is soluble in the mixed-micelle phase represents the total

bioaccessibility. The bioaccessibility of AST in the MRP-AKO emulsion was evaluated

following the simulation of GIT digestion.

Slide 15: Results and Discussions

Notes: At varying reaction periods, The AKO emulsions stabilised by γ-car-CSCP MRPs had

their peroxide and TBAR values measured.

Secondary along with primary oxidation byproducts that formed after 30 days of light-protected

preservation at 25 °C were used to assess the oxidation resistance of the eight emulsions. The

picture illustrates how the hydroperoxide concentration in the control and four emulsions grew

with increasing oxidation time, suggesting a progressive decline in the oil's oxidative stability.

One of the markers used to identify lipid secondary oxidation is TBARS findings.

Slide 16: Conclusion

Notes: The maximum following a 10-day incubating period with DG, MRPs released the FFA

from the AKO emulsion at 25.58% as well as 28.48% of the maximum bioavailability of AST in

the MRPs-AKO emulsion from the gel that MRPs formed was shown by in vitro digestibility.

The creation of emulsified foods containing health-promoting polyunsaturated fats may be

impacted by this research.

SLIDE 17: References

Cui, F., Wang, Q., Han, L., Wang, D., Jiànróng Lǐ, Li, T., & Li, X. (2023). Effect of Maillard conjugates of

peptides and polydextrose on Antarctic krill oil emulsion stability and digestibility. LWT, 179, 114648–

114648. https://doi.org/10.1016/j.lwt.2023.114648
 Fu, J., Song, L., Guan, J., Sun, C., Zhou, D., & Zhu, B. (2021). Encapsulation of Antarctic krill oil in yeast

cell microcarriers: Evaluation of oxidative stability and in vitro release. Food Chemistry, 338, 128089.

https://doi.org/10.1016/j.foodchem.2020.128089

Han, L., Zhai, R., Shi, R., Hu, B., Yang, J., Xu, Z., Ma, K., Li, Y., & Li, T. (2024). Impact of cod skin

peptide-ι-carrageenan conjugates prepared via the Maillard reaction on the physical and oxidative stability

of Antarctic krill oil emulsions. Food Chemistry: X, 21, 101130.

https://doi.org/10.1016/j.fochx.2024.101130

Li, Y., Peng, Z., Tan, L., Zhu, Y., Zhao, C., Zeng, Q.-H., Liu, G., Wang, J. J., & Zhao, Y. (2022).

Structural and functional properties of soluble Antarctic krill proteins covalently modified by rutin. Food

Chemistry, 379, 132159. https://doi.org/10.1016/j.foodchem.2022.132159

Slide 18: Thanking Note