2nd Applied Science Research International Conference 2022 (2ndASRIC 2022)

Antioxidant Activity of Spray Dried Edible Bird’s Nest Hydrolysate

Nur Azra Zulkepli, Rosnani Hasham, Zanariah Hashim*

Food and Biomaterial Engineering Research Group, School of Chemical & Energy Engineering, Faculty of Engineering,
Universiti Teknologi Malaysia, 81310 Johor, Malaysia
*Corresponding authorE-mail: zanariahhashim@utm.my

Abstract: Edible Bird’s Nest (EBN) is made up of glycoprotein and can be hydrolysed into EBN hydrolysates
using enzymatic hydrolysis to produce glycopeptides. Drying is one of the essential steps in EBN processing to
increase the shelf-life of the hydrolysed product, hence spray dryer is selected due to the simple, fast and more
cost-effective process. However, drying process also inhibited the physicochemical changes that may significantly
affect the quality features, notably the nutritional content, which is relatively heat-sensitive. In this study, the
antioxidant activity of non-spray dried (EBNLIQUID) and spray dried (EBNPOWDER) EBN hydrolysates were
investigated using DPPH assay at sub-optimal temperature (16 ± 1 °C) and duration of storage (0, 7, 14, 21 and
28 days). As a result, the initial antioxidant activity of EBNPOWDER (55.41%) is significantly higher than EBNLIQUID
(35.01%), while the reduction of antioxidant activity was lower in EBNPOWDER (10.72%) compared to EBNLIQUID
(16.62%) after 28 days of storage. This study demonstrates that drying can improve the stability of EBN
hydrolysate and prolong its shelf life and bioactivities.

Keywords: Edible bird’s nest, antioxidant, enzymatic hydrolysis, spray dry, sialic acid

1. INTRODUCTION
Nowadays, Edible Bird's Nest (EBN) is becoming more popular due to its health benefits, and Chinese
communities have regarded it as one of their most valuable foods for thousands of years [1]. Proteins
(61-67%) and carbohydrates (25-31%), in the form of glycoproteins, make up the majority of EBN.
Proteases may hydrolyze these glycoproteins into glycopeptides, creating enzyme-mediated EBN
hydrolysates with high bioactivities [2]. Moreover, researchers have discovered that two key functional
elements in EBN are sialic acid (SA) and antioxidants where it can contribute to brain development [3],
learning ability [4] and inhibit influenza virus infection [5].
According to ORAC, FRAP, ABTS and DPPH tests, it was previously discovered that EBN
hydrolysate had more antioxidant activity than unhydrolyzed EBN extract [3]. In this study, due to its
ability to detect a compound's capacity to operate as a free radical scavenger or hydrogen donor, as well
as to assess the antioxidant activity of foods, the DPPH test was chosen.
However, hydrolyzed EBN in liquid form is highly unstable and must be stored at lower
temperatures. Thus, drying process is one of the essential steps in EBN processing. Unfortunately, this
process also inhibited physicochemical changes that can significantly impair the qualitative aspects
[6][7]. Thus, it suggests that an essential factor to take into account is the stability of SA and
antioxidants during the drying of EBN. Therefore, the aim of this study is to investigate the stability of
antioxidant activity for both non-spray dried EBNLIQUID and spray dried EBNPOWDER by using DPPH
assay at sub-optimal temperature (16 ± 1 °C) and duration of storage (0, 7, 14, 21 and 28 days).

2. METHODOLOGY
2.1 Preparation of EBNLIQUID
EBN was hydrolysed according to [2]. Prior to spray drying, 100 mL of the liquid hydrolysate was
mixed with 2.985 g of maltodextrin and 0.015 g of xanthan gum (dissolved in 100 mL distilled water)
which act as carrier agents to assist the spray drying process. The mixture (EBNLIQUID) was constantly
and thoroughly stirred using a homogenizer to ensure homogeneity.

2.2 Drying of EBN Hydrolysates

The mixture of EBNLIQUID was spray-dried using a spray dryer (Lab-Plant Spray Drier SD-04) and the

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flow rate was maintained at 100 mL/h using a peristaltic pump (Masterflex) with an inlet temperature
of 180 ± 2 °C and an output temperature of 90 ± 2 °C to become EBNPOWDER.

2.3 DPPH Assay

Experimental procedures were performed according to the methods in [2]. 0.05 mmol L-1 of methanolic
DPPH solution and 0.5 mL of EBNLIQUID were mixed. A microplate spectrophotometer was used to
detect the absorbance at 517 nm after 30 minutes in the dark. All experiment were repeated with the
storage time of 7, 14, 21 and 28 days at temperature of 16 ± 1 °C for both samples (EBNLIQUID and
EBNPOWDER). For analysis of EBNPOWDER, 1 mg of sample were dissolved in 100 mL of distilled water.
Using equation (1), the antioxidant activities were estimated as a percentage inhibition of DPPH.

Abs of Control − Abs of Sample

DPPH Inhibition (%) = × 100
Abs of Control
(1)
3. RESULTS AND DISCUSSION
As DPPH assay was performed to evaluate antioxidant's scavenging capacity towards DPPH, sample
antioxidant's degree of discolouration shows scavenging ability [8]. The colouring of antioxidants is
caused by the interaction of a stable free radical, α,α-diphenyl-β- picrylhydrazyl (dark violet hue), with
an antioxidant, resulting in a discoloured α-diphenyl-β- picrylhydrazine [9]. In this study, the
antioxidant activities of EBNLIQUID and EBNPOWDER were investigated following storage at 16 ± 1 °C
and duration of 0, 7, 14, 21 and 28 days (Figure 1). As a result, the initial antioxidant activity (Day 0)
of EBNPOWDER (55.41%) is significantly higher (p<0.05) than that of EBNLIQUID (35.01%). Although
there was a slight increase (p>0.05) in the DPPH % inhibition of EBNPOWDER at Day 7, the antioxidant
activities of both samples showed a decreasing trend with storage time. After 28 days of storage, it was
observed that the antioxidant activities of EBNPOWDER and EBNLIQUID reduced to 49.47% and 29.19%,
respectively, with EBNPOWDER (10.72%) exhibited a lower reduction compared to EBNLIQUID (16.62%).

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DPPH (% INHIBITION)

55
50
45
40
35
30
25
0 7 14 21 28
TIME (DAYS)

EBNPOWDER EBNLIQUID

Figure 1. DPPH (% inhibition) for EBNLIQUID and spray dried EBN (EBNPOWDER) stored at 16 ± 2 °C
for 0, 7, 14, 21 and 28 days.

As described by Tonon [10], as water activity increased, antioxidant activity decreased, which is
associated to the greater degradation of antioxidative compounds that exist in these conditions. Gan and
Hui [9] found that the release of bound phenolic compounds, the partial degradation of lignin, which
may liberate phenolic acids, and the heat destruction of sialic acids are the three most likely reasons for
the loss in total antioxidant capability. As found in several antioxidative peptide sequences, the presence
of sialic acid appears to play a crucial function regardless of its location, which has led to its enhanced
radical scavenging capacity. However, it is important to note that in this study, the storage temperature
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was relatively low to avoid the EBNLIQUID from going stale, and the effect of other temperatures has not
been investigated. It is expected that at higher temperatures, the EBNLIQUID will show even higher % of
degradation of antioxidants.

4. CONCLUSION
In conclusion, this study shows that both EBN extracts (EBNLIQUID and EBNPOWDER) might be regarded
as a source of natural antioxidants and can be beneficial for human health. From the results obtained, it
can be demonstrated that EBNPOWDER has a higher antioxidant activity and lower reduction with time
compared to EBNLIQUID, and thus is more suitable for prolonged storage, which indicates that drying
has a great impact on the stability of EBN hyrolysate. Future studies include the evaluation of other
bioactive components in EBN hydrolysates including sialic acid and determining the effect of drying
on other functional properties of EBN.

ACKNOWLEDGMENTS
The research has been carried out under Research Excellence Consortium 2020 (JPT(BKPI)1000/016/018/25(51),
Novel Food Resources Consortium) grant provided by Ministry of Higher Education of Malaysia.

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