Papain Enzyme Assisted Extraction of Virgin Coconut Oil as Candidate In-House Reference Material
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Extraction of VCO
2.3. Physicochemical Analyses of VCO
2.4. Antibacterial Activity of VCO
2.5. Homogeneity Test
3. Results and Discussion
3.1. VCO Extraction
3.2. Determination of Microbial Contamination
3.3. Homogeneity and Stability Tests
3.4. Antibacterial Activity Test
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Factor | Value | Water Content (%) | Free Fatty Acid (%) | Peroxide Number (meq/kg) | Iodine Number (g iodine/100 g) |
---|---|---|---|---|---|
Enzyme to Oil ratio (g/L) | 0 | 0.99 ± 0.12 | 0.24 ± 0.01 | 0.20 ± 0.01 | 7.36 ± 0.04 |
0.2 | 1.01 ± 0.12 | 0.26 ± 0.03 | 0.20 ± 0.01 | 8.13 ± 0.04 | |
0.4 | 1.01 ± 0.12 | 0.30 ± 0.03 | 0.20 ± 0.01 | 8.36 ± 0.06 | |
0.6 | 1.01 ± 0.12 | 0.30 ± 0.04 | 0.20 ± 0.01 | 8.17 ± 0.01 | |
0.8 | 1.01 ± 0.12 | 0.24 ± 0.01 | 0.20 ± 0.01 | 8.45 ± 0.02 | |
1.0 | 1.02 ± 0.12 | 0.28 ± 0.01 | 0.20 ± 0.01 | 8.22 ± 0.09 | |
Volume of water (L/g) | 100 | 1.01 ± 0.22 | 0.23 ± 0.01 | 0.36 ± 0.16 | 7.55 ± 0.01 |
200 | 1.01 ± 0.24 | 0.31 ± 0.09 | 0.18 ± 0.01 | 7.73 ± 0.01 | |
300 | 1.01 ± 0.21 | 0.31 ± 0.18 | 0.18 ± 0.01 | 7.91 ± 0.05 | |
400 | 1.01 ± 0.23 | 0.35 ± 0.06 | 0.18 ± 0.01 | 7.72 ± 0.03 | |
500 | 1.01 ± 0.18 | 0.38 ± 0.12 | 0.18 ± 0.01 | 7.49 ± 0.02 | |
600 | 1.01 ± 0.20 | 0.42 ± 0.08 | 0.18 ± 0.01 | 7.51 ± 0.01 | |
Tempeature of water (°C) | 30 | 1.00 ± 0.12 | 0.31 ± 0.02 | 0.18 ± 0.01 | 8.26 ± 1.02 |
40 | 1.00 ± 0.13 | 0.27 ± 0.05 | 0.18 ± 0.01 | 8.21 ± 1.66 | |
50 | 1.00 ± 0.12 | 0.35 ± 0.06 | 0.18 ± 0.01 | 8.07 ± 1.78 | |
60 | 1.00 ± 0.12 | 0.27 ± 0.05 | 0.18 ± 0.01 | 8.33 ± 0.30 | |
70 | 0.99 ± 0.13 | 0.19 ± 0.04 | 0.18 ± 0.01 | 8.23 ± 0.97 | |
Extraction stage | 1 | 0.99 ± 0.13 | 0.35 ± 0.06 | 0.18 ± 0.01 | 8.23 ± 0.01 |
2 | 1.00 ± 0.12 | 0.38 ± 0.12 | 0.18 ± 0.01 | 7.65 ± 0.04 | |
3 | 0.96 ± 0.14 | 0.27 ± 0.05 | 0.18 ± 0.01 | 7.76 ± 0.03 | |
4 | 0.99 ± 0.13 | 0.27 ± 0.0.5 | 0.18 ± 0.01 | 7.81 ± 0.01 | |
5 | 0.98 ± 0.12 | 0.35 ± 0.06 | 0.18 ± 0.01 | 8.15 ± 0.05 |
Methods | Yield (%) | Water Content (%) | Free Fatty Acid (%) | Peroxide Number (meq/kg) | Iodine Number (g iodine/100 g) | Reference |
---|---|---|---|---|---|---|
Papain | ||||||
Papain at optimum conditions | 24.3 | 0.98 | 0.35 | 0.18 | 8.15 | [11] |
Papain with freezing and thawing | 23 | 0.014 | 0.30 | 0.69 | [12] | |
Papain | 0.11 ± 0.01 | 0.35 ± 0.01 | 4.26 ± 0.05 | [16] | ||
29.8 | 0.18 | 0.22 | [11] | |||
Peel of papaya | 12.7 ± 1.1 | 0.14 ± 0.00 | 0.17 ± 0.00 | 0.57 ± 0.01 | 8.72 ± 0.12 | [10] |
Seed of papaya | 12.5 ± 0.5 | 0.14 ± 0.01 | 0.15 ± 0.01 | 0.60 ± 0.00 | 8.23 ± 0.04 | |
Enzyme | ||||||
Protease | 0.12 ± 0.01 | 0.53 ± 0.14 | 7.62 ± 0.14 | [22] | ||
Bromelain | 24.26 ± 0.06 | 0.34 ± 0.01 | 0.29 ± 0.03 | |||
Fermentation | ||||||
Natural fermentation | 0.13 ± 0.06 | 0.53 ± 0.15 | 2.8 ± 0.03 | 0.91 ± 0.03 | [4] | |
Saccharomyces cerevisiae | 12.40 | 0.04 ± 0.01 | 0.31 ± 0.11 | 0.01 ± 0.00 | 2.44 ± 0.20 | [16] |
0.06 ± 0.00 | 0.29 ± 0.02 | 4.30 ± 0.07 | [23] | |||
Bacillus licheniformis | 0.15 ± 0.01 | 0.78 ± 0.08 | 7.26 ± 0.13 | [22] |
Sample | Dilution | Medium | Incubation | Observation | |||
---|---|---|---|---|---|---|---|
Temperature (°C) | Time (Hours) | I | II | Total | |||
VCO | 10−1 | Plate count agar | 35–37 | 24–48 | 0 | 0 | 0 |
VCO | 10−2 | Plate count agar | 35–38 | 24–49 | 0 | 0 | 0 |
Control | 10−1 | Plate count agar | 35–39 | 24–50 | 0 | 0 | 0 |
Control | 10−2 | Plate count agar | 35–40 | 24–51 | 0 | 0 | 0 |
Blank | 35–41 | 24–52 | 0 | 0 | 0 | ||
Medium | Plate count agar | 35–42 | 24–53 | 0 | 0 | 0 | |
Medium + Diluent solution | Plate count agar-peptone dilution fluid | 35–43 | 24–54 | 0 | 0 | 0 | |
Total plate count | Plate count agar-lactose broth | 35–44 | 24–55 | 0 | 0 | 0 |
Parameter | Mean Square between (MSB) | Mean Square within (MSW) | Ftest |
---|---|---|---|
Free fatty acid (%) | 0.0004 | 0.0001 | 3.0146 |
Peroxide number (meq/kg) | 0.4824 | 0.0614 | 7.8614 |
Iodine number (mg iodine/100 g) | 0.4417 | 0.0551 | 8.0126 |
Parameter | Sample 1 | Sample 2 | |
---|---|---|---|
Free fatty acid (%) | |||
Q1 | 0.1532 | 0.1533 | 0.1533 |
Q3 | 0.2013 | 0.1917 | 0.2003 |
IQR | 0.0482 | 0.0383 | 0.0470 |
nIQR (0.7413 × IQR) | 0.0357 | 0.0284 | 0.0349 |
0.3 × nIQR | 0.1071 | 0.0852 | 0.1046 |
0.0544 | 0.0504 | 0.0509 | |
0.0544 < 0.1071 | 0.0504 < 0.0852 | 0.0509 < 0.1046 | |
Peroxide number (meq/kg) | |||
Q1 | 1.9956 | 2.7906 | 2.5447 |
Q3 | 2.7972 | 3.5888 | 2.7958 |
IQR | 0.8016 | 0.7982 | 0.2511 |
nIQR (0.7413 × IQR) | 0.5942 | 0.5917 | 0.1862 |
0.3 × nIQR | 1.7827 | 1.7750 | 0.5585 |
−0.0392 | 0.9574 | 0.4591 | |
0.0392 < 1.7827 | 0.9574 < 1.7750 | 0.4591 < 0.5585 | |
Iodine number (mg iodine/100g) | |||
Q1 | 7.8283 | 7.9354 | 7.9173 |
Q3 | 8.0926 | 8.0612 | 8.0822 |
IQR | 0.2643 | 0.1258 | 0.1649 |
nIQR (0.7413 × IQR) | 0.1959 | 0.0932 | 0.1222 |
0.3 × nIQR | 0.5878 | 0.2797 | 0.3667 |
1.9507 | 2.0053 | 1.9780 | |
1.9507 > 0.5878 | 2.0053 > 0.2797 | 1.9780 > 0.3667 |
Tested Bacteria | Inhibition Zone for Sample (mm) | |||
---|---|---|---|---|
VCO | VCO-2X | VCO-4X | VCO-8X | |
E. coli | 18.5 | 17 | 11 | 10.5 |
S. aureus | 10 | 12.5 | 10 | 9 |
P. acne | 12 | 10.5 | 9 | 9 |
P. aeruginosa | 10 | 10 | 10 | 10 |
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Yulirohyami; Hidayat, H.; Wijaya, A.R.; Fatimah, I. Papain Enzyme Assisted Extraction of Virgin Coconut Oil as Candidate In-House Reference Material. Processes 2022, 10, 315. https://doi.org/10.3390/pr10020315
Yulirohyami, Hidayat H, Wijaya AR, Fatimah I. Papain Enzyme Assisted Extraction of Virgin Coconut Oil as Candidate In-House Reference Material. Processes. 2022; 10(2):315. https://doi.org/10.3390/pr10020315
Chicago/Turabian StyleYulirohyami, Habibi Hidayat, Aprisilia Rizky Wijaya, and Is Fatimah. 2022. "Papain Enzyme Assisted Extraction of Virgin Coconut Oil as Candidate In-House Reference Material" Processes 10, no. 2: 315. https://doi.org/10.3390/pr10020315