HALAL INGREDIENTS
enzyme-treated extracts were used as new halal cosmetic materials. The production process adheres to the standards of LPPOM MUI (The Assessment Institute for Foods, Drugs, and Cosmetics of Majelis Ulama Indonesia). The carrot-derived ingredients extracted using plant-based enzymes and the Halal cosmetic creams have received Halal certification from LPPOM MUI (Certification No. 00150083580617).
ABTS radical scavenging activity Radicals produced using 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid, ABTS) and potassium persulfate were reacted with the test ingredient for 10 minutes, after which absorbance was measured at 732nm wavelength. The ABTS radical scavenging activity was evaluated based on the reduction in absorbance between the control groups and the sample solutions. The ABTS scavenging activity of the new
carrot-derived enzyme-extracted Halal ingredients, such as papain or bromelain extract, increased according to the concentration. The bromelain-treated extract exhibited superior ABTS radical scavenging activity compared to the papain-treated extract up to a concentration of 10%; however, the two extracts exhibited similar ABTS radical scavenging activity beyond this concentration (refer to Fig 1).
Superoxide dismutase like activity Superoxide dismutase (hereinafter, SOD) is an antioxidant enzyme that produces hydrogen peroxide by reacting with harmful superoxide anion radicals in our body. The produced hydrogen peroxide is subsequently converted into water and oxygen molecules by catalase to protect our body from free radicals. However, the SOD protein is weak to heat and alkali conditions due to its relatively large molecular weight. As such, several investigations on SOD-like activity are underway with the goal of overcoming the aforementioned disadvantages. SOD-like activity measures the amount of produced pyrogallol, which catalyzes the reaction that converts active oxygen species to hydrogen peroxide (H2
O2 );
thus, it can indicate the antioxidant activity of a superoxide. SOD-like activity is measured through the
Marklund-Marklund method. Tris-HCl buffer was added to each sample solution and reacted at 25°C for 10 minutes. Afterwards, the reaction was halted by adding 1N HCl. The absorbance of the oxidised pyrogallol in the reaction solution was measured at 420nm wavelength, and the decreases in absorbance between the control and reacted solutions of the sample were calculated.
Carrot papain 5.0 ■ Carrot bromelain 5.0 ■
100 80 60 40 20 0
63
1
50
75 New carrot halal ingredient concentration (%) Figure 3: Collagenase inhibitory activities of the new carrot Halal ingredients. Carrot papain 5.0 ■ Carrot bromelain 5.0 ■
60 50 40 30 20 10 0
1 50 75 New carrot halal ingredient concentration (%)
Figure 4: Elastase inhibitory activities of the new carrot Halal ingredients. The superoxide dismutase (SOD)-like
activity of the Halal carrot extracts increased in a dependent manner according to the concentration of the extract. The two Halal carrot extracts exhibited similar SOD activities up to the 40% treatment group; beyond this concentration, the papain-treated extract showed superior SOD-like activity compared to the bromelain-treated extract (refer to Fig 2).
Collagenase inhibitory activity Collagenase inhibitory activity was measured by following the method of Wüunsch and Heindrich. In 0.1M Tris-HCl buffer (pH 7.5) containing 4mM CaCl2, the test substance was mixed with a substrate dissolved with 4-phenylazobenzyloxycarbonyl-Pro-Leu- Gly-Pro-D-Arg, after which collagenase was added and reacted at room temperature. Citric
100
20
acid was then added to the mixture to halt the reaction. Afterwards, ethyl acetate was added and mixed to separately obtain the supernatant. Subsequently, the absorbance was measured at 320nm wavelength. The collagenase inhibitory activity of the
Halal carrot extracts increased in a dependent manner according to the concentration of the extract. Both extracts exhibited optimal collagenase inhibitory activity in the 75% treatment group (refer to Fig 3).
Elastase inhibition assay 60µl of distilled water, 10µl of 50mM Tris-HCl buffer (pH 8.0), and 10µl of 10mM substrate (N-succinyl-(Ala)3-4- nitroanilide) was mixed and reacted at 37°C for five minutes. 10µl of elastase (1unit/ml) was then added to the substrate and sample mixture and
Figure 5: Effects of the new carrot Halal ingredients on UVB-irradiated HDF cell line viability.
www.personalcaremagazine.com July 2021 PERSONAL CARE
Elastase inhibition (%)
Collagenase inhibition (%)
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