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64 HALAL INGREDIENTS


the absorbance was measured at 415ml wavelength. The Halal carrot extracts exhibited elastase inhibitory activity starting from the 50% treatment group, and the bromelain- treated group exhibited superior inhibitory activity compared to the papain-treated group (refer to Fig 4).


Total polyphenol content assay The total polyphenol content was measured according to the Folin-Denis method. Distilled water was added to the extract sample and 0.5ml of this solution was placed in a test tube. A phenol reagent was then added and thoroughly mixed, and the mixture was left to sit for five minutes. Afterwards, a saturated Na2


CO3 solution was added and


reacted for one hour. 1N HCl (0.1ml) was then added and the absorbance was measured at 640m wavelength. The total phenol content was converted to a standard curve that uses tannic acid. The total polyphenol content of


the Halal carrot extracts increased in a dependent manner according to the enzyme concentration. The total polyphenol content of the carrot-papain-treated extract increased by 16% from 1.64% to 1.91% as the enzyme concentration increased from 2.5% to 5.0%, and the carrot-bromelain-treated extract showed similar increases (refer to Table 1).


Cell viability Human Dermal Fibroblast (HDF) cells were seeded with a density of 0.8 x 105


irradiation. The cell viability effects of the carrot


enzyme-treated extract were evaluated by observing its effects on the viability of UVB- irradiated HDF cell lines. Compared to the control group, which was not irradiated with UVB, the UVB-irradiated group exhibited a decrease in the number of cells as well as apoptosis. However, the UVB-irradiated + new papain-extracted ingredient and the UVB-irradiated +new bromelain-extracted ingredient groups both inhibited apoptosis and resulted in the number of cells decreasing by a lesser degree when treated to a concentration of 50µl/ml (refer to Fig 5).


100 80 60 40 20 0


*** *** *** *** 56.56 50.06 30.98 31.16 31.18 31.10 43.25 30.88 38.16 31.7 cells/well


and treated with the Halal carrot extracts. Cell viability was observed upon UV-B (15mJ/cm2


)


TABLE 1: TOTAL POLYPHENOL CONTENTS OF THE NEW CARROT HALAL INGREDIENTS Groups (%)


1st


Carrot papain 2.5 Carrot papain 5.0 Carrot bromelain 2.5 Carrot bromelain 5.0


1.58 1.87 1.61


1.86


* Carrot papain 5.0 – Carrot extracts treated with 5.0% papain Carrot bromelain 5.0 – Carrot extracts treated with 5.0% bromelain


Moisturising effects of creams with animal- based collagen and the new carrot-derived enzyme-extracted Halal ingredients A human trial test was conducted with a group of 20 women with an average age of 31 years to observe the lasting 24-hour skin moisturising effects. The test was conducted by drawing five 3 × 3cm squares at 1.5 cm intervals on the flexor surfaces of the left and right forearms 5 cm from the wrist, and the test samples were applied on these sites. After the test subjects waited for 30 minutes in a waiting room with controlled temperature and humidity conditions (20∼24°C, 50∼60% RH), each test site was measured three times. The hydration level was measured before any product was applied (0 hours), and a Corneometer (CM825, Courage & Khazaka, Germany) was used with the same method to measure the hydration level 1, 3, 6, and 24 hours after applying the products. Animal collagen and the two new carrot-


derived enzyme-extracted Halal ingredients were implemented into creams to evaluate the skin moisturising capabilities of the substances over time. The creams with the new carrot- derived enzyme-extracted Halal ingredients exhibited superior moisturising capabilities compared to the cream containing animal- based collagen for all evaluation time intervals. The two creams with the new carrot-derived enzyme-extracted Halal ingredients increased skin moisture by 4% or more after one hour of application on the skin and by 7.5% or more after six hours of application compared to the cream containing animal-based collagen. As a result, this study provides evidence


that the creams containing the new carrot- derived enzyme-extracted Halal ingredients possess superior skin moisturising capabilities


0hr 1hr 3hr Hours Not treated ■ Collagen cream ■ 6hr 24hr


100 80 60 40 20 0


*** *** *** *** 58.99 53.52 30.82 31.16 31.18 31.10 46.65 30.88 40.16 31.7


compared creams containing animal-based collagen (refer to Fig 6).


Conclusions To develop new ingredients for Halal cosmetics that can replace animal-based collagen, Chemland used the plant-based enzymes papain and bromelain to extract ingredients from agricultural products such as carrots, oyster mushrooms, mung beans, and red ginseng. These ingredients have received LPPOM MUI new Halal ingredient certification from Majelis Ulama Indonesia. Among the certified ingredients, this paper analyzed the ABTS radical scavenging activity, superoxide dismutase activity, collagenase and elastase inhibition assays, and total polyphenol content of Halal carrot-derived enzyme extracts. Furthermore, Halal creams were produced and the moisturising effects were studied through clinical trials. As a result of the research, the new carrot-


derived enzyme-extracted Halal ingredients showed superior antioxidant and anti-wrinkle activities regardless of whether the ingredient was treated with papain or bromelain. According to an analysis of the effects of UVB irradiation (15 mJ/cm2


) on the growth of cell


lines, both ingredients inhibited apoptosis and resulted in the number of cells decreasing by a lesser degree regardless of the treatment enzyme. The clinical trial results also showed that creams with the new carrot-derived enzyme-extract Halal ingredients possessed 5% or higher moisturising capabilities. The aforementioned results highlight the potential of the new carrot-derived enzyme-extracted Halal ingredients as superior cosmetic ingredients for the Halal cosmetics market.


0hr 1hr 3hr Hours Not treated ■ Carrot-papain cream ■ 6hr 24hr


100 80 60 40 20 0


*** *** *** *** 59.37 31.34 31.16 31.18 31.10 30.88 53.69 46.93 2nd


1.67 1.93 1.65 1.94


3rd


1.66 1.92 1.66 1.92


mean ± SD


1.64 ± 0.05 1.91 ± 0.03 1.64 ± 0.03 1.91 ± 0.04


PC


40.52 31.7


0hr


1hr


3hr Hours Not treated ■ Corrot-bromelain cream ■


Figure 6: Moisturizing effects of a cream containing animal-based collagen and two creams containing the new carrot Halal ingredients. Student t-test *** P<0.05


PERSONAL CARE July 2021 www.personalcaremagazine.com


6hr


24hr


Moisturizing value (A.U.)


Moisturizing value (A.U.)


Moisturizing value (A.U.)


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