Citation
Bahari, Aqilah Noor
(2021)
Development of halal hydrolysates-based nanocosmeceutical formulation from Actinopyga lecanora.
Doctoral thesis, Universiti Putra Malaysia.
Abstract
Actinopyga lecanora is classified among the edible species of sea cucumber. Its
hydrolysates, which obtained from enzymatic hydrolysis was reported to contain
relatively high antioxidant activities, mostly applied in food industries. Actinopyga
lecanora hydrolysates (ALH) is halal depending on its source (marine animal),
extraction process and the safety measurement. Even though ALH have high
potential of antioxidant activities, which is essential in skin care products, no work
has been reported specifically on ALH as an actives ingredient. Likewise, no
related antityrosinase activity of ALH has been recorded. Therefore, to diversify
the usage of ALH in the cosmeceutical application, nanoemulsion containing
ALH was developed, particularly to alleviate skin aging and to improve skin
pigmentation.
This research consists of two parts; Part A is the optimization of hydrolysis
conditions of ALH, while Part B covers the formulation of nanoemulsion
containing ALH obtained from Part A. In Part A, ALH was hydrolysed from raw
sample of Actinopyga lecanora using papain enzyme. The best conditions
selected from the optimization were at pH 5.00, 70°C of reaction temperature, 9
h of hydrolysis time and 1.00% E/S ratio with the ALH having 55.00% of DH,
46.30% of DPPH radical scavenging activity and 108.20 Fe2+μg/mL of FRAP
activity.
The amount of hydrophobic amino acid (286.40 mg/g sample) was higher than
hydrophilic amino acid (253.60 mg/g sample), indicated that the hydrophobic
amino acid was responsible for the antioxidant and antityrosinase activities. SEM
image of ALH pictured a smooth structures with pores as the protein was
degraded into smaller peptides.
In Part B, nanoemulsion formulation containing ALH was optimized using Doptimal
mixture design. The optimum compositions were at 10.00% of safflower
oil: argan oil (ratio 9:1), 1.00% of ALH, 13.75% of Tween 80: Span 80 (ratio 8:2)
surfactants, 0.87% of xanthan gum, 73.69 % of water and 0.70% of phenonip.
The actual particle size obtained was 139.80 nm, with the acceptable
polydispersity index (0.302) and zeta potential (-27.50 mV). High conductivity of
the nanoemulsion was observed (1311 μS/cm) with pH 5.38 and displayed a
pseudoplastic behaviour. Morphology analysis disclosed a spherical shape of
the nanoemulsion particles, monodispersed without any aggregation. Moreover,
the nanoemulsion showed good stability within 28 days when stored separately
at different temperatures (4 and 25°C). However, for long term storage, the
suggested storage condition was at 4°C due to the unchanged of physical
appearances (colour and odour) after 28 days.
In vitro permeation study revealed that the permeability of ALH was improved to
66.48% of ALH released after 8 h of application. The kinetic released of ALH
followed first-order model, in which the released was based on the concentrationgradient
pattern. In vitro cytotoxicity test of all samples against normal 3T3
fibroblast cell recorded IC50 values more than 500 μg/mL, proved that the
optimized nanoemulsion is safe, halal and suitable for topical application.
Apart from that, an improved in antioxidant activity was shown after incorporating
ALH to the nanoemulsion. The DPPH radical scavenging activity of
nanoemulsion containing ALH had a significant difference (p<0.05) at
concentration of more than 5000 μg/ml compared to the blank nanoemulsion.
The FRAP activity of both samples also showed a significant difference (p<0.05)
at every sample concentration. Similar pattern was found for antityrosinase
activity whereby the optimized nanoemulsion containing ALH displayed a
significant difference (p<0.05) in tyrosinase inhibition activities for both
substrates, L-DOPA (43.20%) and L-tyrosine (46.60%). Overall, the
nanoemulsion containing halal hydrolysates from A. lecanora was successfully
formulated and have a full potential to be applied in cosmeceutical industries.
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