1.

IDENTIFICATION OF THE ARTICLE

Title: Effects of liquid nitrogen quick freezing on polyphenol oxidase and
peroxide activities, cell water states and epidermal microstructure of wolfberry
Authors: Zhiwei Zhu, Wenhuang Luo, Da-Wen Sun
Published in: School of Food Science and Engineering, South China University
of Technology, Guangzhou, China
Publication date: 2018
Produced by: LWT - Food Science and Technology

2. SUMARY
To better preserve the quality and nutritional value of wolfberry, the freezing
effect by spraying liquid nitrogen at 60 °C ± 2 °C (60 °C), 80 °C ± 2 °C (80 °C)
and 100 °C ± 2°C (100°C). on wolfberry, compared to ice formation with air blast
at 40 °C ± 1 °C and an aerodynamic velocity of 0.75 m/s. The results show that
the freezing time to pass through the region of maximum ice crystal formation is
450 s, 150 s, 100 s and 70 s at 60 °C, 80 °C and 100 °C, respectively. at 80 °C
had better shape, lower peroxidase activity, more similar water distribution to
fresh samples and less damage to the internal epidermal cell structure.

3. KEYWORDS
Wolfberry, Liquid nitrogen quick freezing, Water distribution, Epidermal structure
Freezing characteristic

4. PROBLEM OF INVESTIGATION
Lycium chinense, also known as Lycium barbarum, is a berry native to Asia and
grown mainly in arid and semi-arid areas which contains many biologically active
substances, including carotenoids, polysaccharides, polyphenols and nitrogen
compounds with strong antioxidant and anti-aging properties. After harvest the
wolfberry usually have a very short shelf life at room temperature and change
color and flavor after 3 to 5 days. When stored at 4 ° C, the wolfberry loses water
and shrinks in 23 weeks. Therefore, the most common method for storage is
drying. But some during drying functional nutrients such as amino acids,
carotenoids and polysaccharides are inevitably lost, so an efficient way to
maximize nutrient retention of wolfberries during storage is needed. Liquid
nitrogen spray freezing is a method of flash freezing foods using instant liquid
nitrogen. Compared with the conventional freezing method, it has the
advantages of high heat transfer coefficient, fast freezing speed, short freezing
time and fine ice crystal formation. If the freezing rate is high enough, the regions
where ice crystals are maximally formed in the aqueous solution can pass very
quickly without tissue crystallization. A freezing rate of 106 K/s is normally
required to achieve a complete transition to the free state. However, for foods
with high moisture content, it is difficult to achieve such a high freezing rate.
Therefore, the use of partial transition glazing is a more common practice by
damaging the tissue structure by recrystallization during frozen storage, which
will result in further loss of drip, nutrients such as carbohydrates and vitamins,
and even reduce the quality of the food. For foods stored below the glass
transition temperature, much of the nutrient loss can be controlled, and freezing
with liquid nitrogen is one of the most common techniques to achieve a partial
glass transition in the food freezing process. . However, little is known about
freezing wolf by spraying liquid nitrogen.

5. OBJECTIVES
To investigate the effect of liquid nitrogen spray freezing of wolfberry on color,
polyphenoloxidase and peroxide activity, water status distribution and epidermal
microstructure at different temperatures.

6. METHODOLOGICAL APPROACH: METHODOLOGY USED FOR THE

DEVELOPMENT OF THE INVESTIGATION
The methodology used was the application of quantitative research techniques to
determine the chemical changes produced during the heat treatment of milk. All
experiments were repeated at least three times and the results were expressed
as mean ± standard deviation (mean ± std). Experimental data were statistically
analyzed using SPSS software. A one-way analysis of variance (ANOVA) was
performed on the data.

7. TECHNOLOGICAL CHARACTERISTICS

Wolfberry harvesting

Wild wolfberry fruit contains about 80% water, and the main chemical
components include Lycium barbarum polysaccharide, carotenoids, alkaloids,
and vitamin C. Fresh wolfberry berries were harvested in October 2018 from
Zhongwei Farm, Ninh Province, Ha, China. After harvesting, the samples were
transported to the laboratory within two days by airmail at low temperature. Lobo
berries were not clearly crushed and the hard pulp was selected for the
experiment. Before the experiment, all wolfberry samples were sorted, and the
mean length and weight of the mean length and weight of the wild wolfberry were
3 ± 0.2 cm and 1.1, ± 0.1 g, respectively. Before testing, the samples were
washed with water and the surface was dried.

Freezing experiment

Wolfberry were stored under refrigeration at 2 ° C. Samples were frozen by

spraying liquid nitrogen at 60 ± 2 ° C, 80 ± 2 ° C and 100 ± 2 ° C in a nitrogen
freezing cabinet,
respectively. For comparison, samples were also frozen in a pneumatic freezer
with an air freezing temperature of 40 ± 1 °C and an air freezing velocity of 0.75
m/s. Wolfberry temperature during freezing was monitored by a thermocouple
inserted into the geometric center of the sample, connected to a data logger.
Data were recorded at a 2 s interval. Freezing continued until the core
temperature reached the experimentally measured partial glass transition
temperature of the wolfberry. Then, the frozen wolfberry is stored at glass
temperature for three weeks. For index analysis, the frozen wolfberry was
thawed at 2 °C for 12 h in the refrigerator.

Measuring color parameters

The color of the samples was determined using a colorimeter. The color
parameters recorded were brightness values (L*), red or green values (a*), and
yellow or blue values (b).

Determination of polyphenol oxidase and peroxidase activities

Polyphenol oxidase and peroxidase activities in fresh and thawed form were
measured using an ultraviolet spectrophotometer. absorbance was measured,
The amount of enzyme causing an absorbance change of 0.01 per minute was
used as the unit of polyphenol oxidase activity. water status distribution Storage
nuclear magnetic resonance can be used to determine the water status, changes
and distribution of fruits from a microscopic point of view. All tests were
performed at room temperature 25 °C.

Observation of the microstructure of the epidermis

The microstructure of Lobo berry epidermis was observed microscopically. A

slicer was used to cut fresh and thawed samples to a thickness of less than 20
μm. The epidermis was then cut into 5 x 5 mm square pieces and sealed with
coverslips. Images were taken at 40x magnification. All tests were performed at
room temperature 25 °C.

8. PRESENTATION OF RESULTS
The glass transition temperature of Lobo berry and the freezing curve in the
glassy state, the molecular thermal kinetic energy is very low, the number of
kinetic units is small and the molecular chain is frozen, which causes little
physical and chemical reaction within the food. Minimal degradation of food
quality was evident. The enthalpy value of the wolfberries increased gradually
with increasing temperature. At temperatures below 42.1 °C, the enthalpy value
changed little and above 42.1 °C, the enthalpy value increased significantly,
there was a tipping point at 39.2 °C, which was determined as the transition
temperature compares the changes in wolfberry core temperature during various
freezing processes. It was observed that the time to cross the zone of maximum
ice crystal formation was approximately 450 seconds, 150 seconds, 100 seconds
and 70 seconds at, Nitrogen 60°C, Nitrogen 80°C and Nitrogen 100°C,
respectively.
Color change
Fresh wolfberry fruit had a bright color, but when frozen it deteriorated and
darkened. At temperatures of 100°C, slight creases were found in the epidermis
of wolfberry. Large ice crystals can form due to the slow freezing rate, and
although fine ice crystals were expected to form, the thermal stress on the
epidermis should be high due to the large difference. Of the freezing
environment and epidermis. In addition, the wax layer on the wolfberry surface is
irreversibly damaged during freezing, which reduces the water-holding capacity
of the wolfberry and causes the epidermis to shrink. They found that the faster
the freezing rate, the more severe the decrease in the activity of
Polyphenoloxidase and peroxidase enzymes.

Changes in the distribution of water conditions.

The moisture contained in wolfberry is mainly distributed in the cell wall, stromal
cells, cytoplasm and vacuole, and moves when frozen.

It could be shown, compared with the relaxation peak at various temperatures of

fresh wolfberry, the peak height and peak width of the water peak of each berry
component changed significantly after freezing. Water peak height decreased
slightly and peak width increased significantly, but there was no significant
change in peak position. Therefore, the ice crystals formed during the water
crystallization process damaged the cellular structures.

In wolfberry, the proportion of free water at 24°C decreased significantly, while

the proportion of bound and bound water increased slightly. The greater the
decrease in free water, the greater the damage to the vacuole.

During freezing, most of the water moved from the vacuole to the cytoplasm and
cell space, and only a part of the water migrated to the cell wall.

9. CONCLUSIONS
Although this study showed that freezing temperature affects the freezing
performance of wolfberries, there is no evidence that the lower the temperature,
the better the freezing performance. Color change, polyphenoloxidase and
peroxidase activity, water transport distribution, and internal microstructure of the
epidermis were compared under various freezing processes. At 100°C with
nitrogen there are the advantages of short freezing times and low
polyphenoloxidase activity. However, compared to 100 °C, 80 °C with nitrogen
produced better organoleptic quality and lower polyphenoloxidase activity, water
distribution more similar to fresh samples, and less damage to cellular structures
within the epidermis. Slow freezing obtained wolfberries with a color close to that
of the fresh sample, but changes in other quality indicators were not desirable.
Therefore, it has been suggested that 80°C with nitrogen is the most suitable for
the wolfberry freezing process.
10. ARTICLE REVIEWER'S COMMENTS (STUDENT)
In the review of the article it is possible to highlight the between freezing
treatments with different mechanisms, since freezing was used by immersing the
wolfberries in nitrogen and in a pneumatic freezing system, the speed of nitrogen
to freeze the food is highlighted.
 REFERENCIAS BIBLIOGRAFICAS

Zhiwei Zhu, Wenhuang Luo, Da-Wen Sun; Effects of liquid nitrogen quick freezing
on polyphenol oxidase and peroxide activities, cell water states and epidermal
microstructure of wolfberry; 2020; Published in:
https://www.sciencedirect.com/science/article/abs/pii/S0023643819312654