Comprehensive Evaluation of Parameters Affecting One-Step Method for Quantitative Analysis of Fatty Acids in Meat
Abstract
:1. Introduction
2. Results and Discussion
2.1. Influence of Reaction Temperature
2.2. Influence of Acid Concentration
2.3. Influence of Moisture Content
2.4. Influence of Solvent Volume and Sample Weight
2.5. Statistical Significance of the Influence of the Different Parameters Investigated
2.6. Influence of the Salt Type
2.7. Repeatability of the Optimised Direct Method for Quantitative Determination of Fatty Acid in Meat from Lamb, Beef and Venison
2.8. Comparative Evaluation Between the Optimised One-Step and Direct Bimethylation Procedures
3. Conclusions
4. Materials and Methods
4.1. Reagents
4.2. Samples
4.3. Experimental Design
4.4. FA Derivatisation using Bimethylation Procedure
4.5. FA Derivatisation using Optimised One-Step Procedure
4.6. FAME Analysis
4.7. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Fatty Acids | P-values | R2-adj Values | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|
- | Individual Effect | Interaction | |||||||||
- | Temperature | Time | Acid Concentration | Volume | Temp * Time | Temp * Acid Concentration | Temp * Vol | Time * Acid Concentration | Time * Vol | Acid Concentration * Vol | |
Total Fatty Acids | <0.001 | <0.001 | <0.001 | 0.005 | <0.001 | 0.052 | 0.002 | 0.237 | 0.052 | 0.212 | 45.4% |
C10:0 | <0.001 | <0.001 | 0.002 | <0.001 | <0.001 | 0.525 | 0.015 | 0.417 | 0.105 | 0.299 | 33.5% |
C12:0 | <0.001 | <0.001 | 0.005 | 0.061 | <0.001 | 0.679 | 0.086 | 0.488 | 0.184 | 0.546 | 26.5% |
C14:0 | <0.001 | <0.001 | <0.001 | 0.022 | <0.001 | 0.067 | <0.001 | 0.284 | 0.042 | 0.199 | 45.4% |
Iso C15 | <0.001 | <0.001 | 0.004 | <0.001 | <0.001 | 0.071 | 0.239 | 0.503 | 0.348 | 0.400 | 42.6% |
Anteiso C15 | <0.001 | <0.001 | <0.001 | <0.001 | <0.001 | 0.066 | 0.036 | 0.346 | 0.031 | 0.558 | 43.5% |
C15:0 | <0.001 | <0.001 | <0.001 | <0.001 | <0.001 | 0.130 | 0.003 | 0.222 | 0.026 | 0.275 | 46.3% |
Iso C16 | <0.001 | 0.129 | 0.008 | 0.366 | 0.049 | 0.408 | 0.125 | 0.754 | 0.101 | 0.914 | 36.5% |
C16:0 | <0.001 | <0.001 | <0.001 | 0.005 | <0.001 | 0.062 | 0.001 | 0.334 | 0.047 | 0.235 | 46.0% |
Iso c17 | 0.041 | <0.001 | <0.001 | <0.001 | <0.001 | 0.019 | <0.001 | 0.328 | 0.059 | 0.223 | 41.2% |
C16:1 | <0.001 | <0.001 | <0.001 | 0.002 | <0.001 | 0.037 | 0.002 | 0.269 | 0.056 | 0.238 | 43.3% |
Anteiso C17 | <0.001 | <0.001 | 0.001 | 0.010 | <0.001 | 0.149 | 0.017 | 0.416 | 0.048 | 0.301 | 38.5% |
C17:0 | <0.001 | <0.001 | <0.001 | <0.001 | <0.001 | 0.063 | 0.001 | 0.310 | 0.050 | 0.219 | 45.9% |
C17:1 | <0.001 | 0.001 | <0.001 | 0.001 | <0.001 | 0.046 | 0.006 | 0.250 | 0.056 | 0.232 | 39.3% |
C18:0 | <0.001 | <0.001 | <0.001 | 0.011 | <0.001 | 0.063 | 0.001 | 0.322 | 0.039 | 0.216 | 47.1% |
C18:1 9t | <0.001 | <0.001 | <0.001 | <0.001 | <0.001 | 0.847 | 0.004 | 0.871 | 0.091 | 0.558 | 65.1% |
C18:1 11t | <0.001 | <0.001 | <0.001 | 0.003 | <0.001 | 0.093 | <0.001 | 0.258 | 0.038 | 0.232 | 45.1% |
C18:1 9c | <0.001 | 0.01 | 0.001 | <0.001 | <0.001 | 0.156 | 0.005 | 0.443 | 0.176 | 0.681 | 37.7% |
C18:1 11c | <0.001 | 0.010 | 0.001 | <0.001 | <0.001 | 0.156 | 0.005 | 0.443 | 0.176 | 0.681 | 33.6% |
C18:2 n6 | <0.001 | 0.077 | <0.001 | 0.111 | <0.001 | 0.005 | 0.003 | 0.260 | 0.053 | 0.177 | 29.2% |
C20:0 | 0.407 | 0.061 | 0.001 | 0.308 | <0.001 | 0.254 | 0.055 | 0.541 | 0.735 | 0.636 | 4.7% |
C18:3 n3 | <0.001 | 0.014 | <0.001 | 0.013 | <0.001 | 0.013 | 0.002 | 0.269 | 0.041 | 0.198 | 33.5% |
CLA 9c, 11t | <0.001 | 0.784 | 0.301 | 0.274 | <0.001 | <0.001 | <0.001 | 0.282 | 0.066 | 0.333 | 38.3% |
C20:4 n6 | 0.695 | 0.821 | 0.001 | 0.031 | <0.001 | 0.003 | 0.039 | 0.123 | 0.271 | 0.084 | 17.5% |
C20:5 n3 | <0.001 | 0.554 | 0.856 | 0.934 | 0.906 | 0.819 | 0.443 | 0.660 | 0.861 | 0.729 | 24.7% |
C22:5 | 0.543 | 0.739 | <0.001 | 0.002 | <0.001 | 0.001 | 0.006 | 0.385 | 0.011 | 0.112 | 23.0% |
C22:6 n3 | 0.053 | 0.143 | <0.001 | 0.067 | <0.001 | 0.028 | 0.056 | 0.412 | 0.196 | 0.093 | 21.8% |
FAs | Na2CO3 | NaCl | ||
---|---|---|---|---|
Mean ± STD | RSD | Mean ± STD | RSD | |
C10:0 | 0.21 ± 0.00 | 1.5 | 0.20 ± 0.00 | 2.4 |
C12:0 | 0.44 ± 0.01 | 1.6 | 0.44 ± 0.01 | 1.9 |
C14:0 | 3.81 ± 0.07 | 1.9 | 3.76 ± 0.07 | 1.9 |
iso C15 | 0.13 ± 0.00 | 2.8 | 0.13 ± 0.00 | 2.7 |
anteiso C15 | 0.19 ± 0.00 | 2.3 | 0.19 ± 0.01 | 3.6 |
C14:1 | 0.10 ± 0.00 | 1.7 | 0.10 ± 0.00 | 2.0 |
C15:0 | 0.45 ± 0.01 | 1.2 | 0.43 ± 0.01 | 2.2 |
iso C16 | 0.16 ± 0.01 | 3.8 | 0.16 ± 0.01 | 4.0 |
C16:0 | 34.26 ± 0.37 | 1.1 | 34.14 ± 0.38 | 1.1 |
iso C17 | 0.54 ± 0.01 | 1.5 | 0.53 ± 0.01 | 1.7 |
C16:1 | 1.60 ± 0.02 | 1.1 | 1.57 ± 0.02 | 1.5 |
anteiso C17 | 0.61 ± 0.01 | 2.2 | 0.59 ± 0.01 | 1.8 |
C17:0 | 1.44 ± 0.01 | 0.8 | 1.40 ± 0.02 | 1.5 |
C17:1 | 0.75 ± 0.00 | 0.6 | 0.73 ± 0.01 | 0.9 |
C18:0 | 24.17 ± 0.26 | 1.1 | 24.05 ± 0.27 | 1.1 |
C18:1 9t | 0.31 ± 0.01 | 1.8 | 0.30 ± 0.01 | 3.1 |
C18:1 11t | 3.77 ± 0.04 | 1.0 | 3.69 ± 0.06 | 1.8 |
C18:1 9c | 56.32 ± 0.63 | 1.1 | 56.28 ± 0.47 | 0.8 |
C18:1 11c | 1.39 ± 0.01 | 0.7 | 1.35 ± 0.01 | 1.0 |
C18:2 n6 | 4.70 ± 0.03 | 0.7 | 4.58 ± 0.04 | 0.9 |
C20:0 | 0.10 ± 0.00 | 1.5 | 0.10 ± 0.00 | 3.4 |
C18:3 n3 | 3.04 ± 0.02 | 0.5 | 2.98 ± 0.06 | 2.0 |
CLA 9c, 11t | 1.72 ± 0.02 | 0.9 | 1.71 ± 0.08 | 4.6 |
C22 | 0.16 ± 0.00 | 1.4 | 0.15 ± 0.04 | 23.0 |
C20:4 n6 | 1.20 ± 0.02 | 1.7 | 1.17 ± 0.02 | 1.6 |
C22:1 | 0.10 ± 0.04 | 4.2 | 0.08 ± 0.01 | 15.9 |
C20:5 n3 | 1.17 ± 0.01 | 1.2 | 1.14 ± 0.01 | 1.2 |
C24:0 | 0.07 ± 0.01 | 11.5 | 0.07 ± 0.01 | 11.1 |
C24:1 | 0.06 ± 0.00 | 7.4 | 0.09 ± 0.02 | 27.3 |
C22:5 | 1.08 ± 0.01 | 1.3 | 1.06 ± 0.01 | 1.3 |
C22:6 n3 | 0.30 ± 0.01 | 2.5 | 0.29 ± 0.01 | 3.5 |
Unreported FA | 10.48 ± 0.88 | 8.4 | 10.28 ± 0.85 | 8.3 |
Sum FA | 154.85 ± 1.83 | 1.2 | 153.55 ± 1.87 | 1.2 |
FA | Lamb Chumps | Lamb Loin | Lamb Shoulder | Beef | Venison | |||||
---|---|---|---|---|---|---|---|---|---|---|
Mean ± STD | RSD | Mean ± STD | RSD | Mean ± STD | RSD | Mean ± STD | RSD | Mean ± STD | RSD | |
C14:0 | 0.84 ± 0.01 | 1.25 | 0.70 ± 0.01 | 1.47 | 8.36 ± 0.27 | 3.18 | 0.64 ± 0.01 | 1.34 | 0.21 ± 0.01 | 4.38 |
C14:1 | 0.03 ± 0.00 | 4.08 | 0.02 ± 0.00 | 5.09 | 0.22 ± 0.02 | 8.77 | 0.18 ± 0.00 | 2.53 | 0.06 ± 0.00 | 3.74 |
C15:0 | 0.13 ± 0.00 | 1.59 | 0.10 ± 0.00 | 2.72 | 1.29 ± 0.04 | 3.39 | 0.13 ± 0.00 | 2.31 | 0.05 ± 0.00 | 5.06 |
C16:0 | 5.61 ± 0.06 | 1.14 | 5.12 ± 0.04 | 0.69 | 42.21 ± 1.22 | 2.89 | 7.50 ± 0.11 | 1.46 | 1.67 ± 0.04 | 2.69 |
C16:1 | 0.28 ± 0.01 | 2.40 | 0.24 ± 0.01 | 2.87 | 2.01 ± 0.06 | 3.03 | 0.90 ± 0.01 | 1.17 | 0.32 ± 0.01 | 3.16 |
C17:0 | 0.30 ± 0.01 | 1.77 | 0.25 ± 0.00 | 1.41 | 2.91 ± 0.09 | 3.07 | 0.31 ± 0.01 | 1.86 | 0.06 ± 0.00 | 5.52 |
C17:1 | 0.14 ± 0.00 | 1.96 | 0.12 ± 0.00 | 2.28 | 1.01 ± 0.03 | 3.10 | 0.25 ± 0.00 | 1.62 | 0.04 ± 0.00 | 7.28 |
C18:0 | 4.44 ± 0.05 | 1.12 | 3.88 ± 0.02 | 0.63 | 35.39 ± 0.92 | 2.61 | 4.47 ± 0.06 | 1.33 | 1.54 ± 0.03 | 1.86 |
C18:1 11t | 0.80 ± 0.02 | 1.89 | 0.69 ± 0.02 | 3.23 | 7.86 ± 0.30 | 3.79 | 0.37 ± 0.01 | 2.09 | 0.12±0.01 | 5.93 |
C18:1 9c | 7.68 ± 0.09 | 1.21 | 7.34 ± 0.06 | 0.81 | 53.06 ± 1.37 | 2.58 | 12.07 ± 0.15 | 1.26 | 1.61 ± 0.04 | 2.46 |
C18:1 11c | 0.34 ± 0.00 | 1.34 | 0.29 ± 0.01 | 4.68 | 1.90 ± 0.09 | 4.61 | 0.55 ± 0.01 | 2.71 | 0.37 ± 0.01 | 1.85 |
C18:2 n6 | 1.59 ± 0.02 | 1.00 | 1.12 ± 0.02 | 1.52 | 4.57 ± 0.18 | 3.96 | 0.68 ± 0.01 | 1.07 | 1.30 ± 0.02 | 1.51 |
C18:3 n3 | 0.88 ± 0.01 | 1.05 | 0.64 ± 0.01 | 1.04 | 3.67 ± 0.11 | 2.95 | 0.42 ± 0.01 | 2.19 | 0.51 ± 0.01 | 2.19 |
CLA 9c, 11t | 0.29 ± 0.01 | 2.09 | 0.28 ± 0.01 | 2.66 | 2.40 ± 0.11 | 4.39 | 0.19 ± 0.01 | 4.20 | 0.04 ± 0.00 | 9.07 |
C20:4 n6 | 0.34 ± 0.00 | 1.00 | 0.27 ± 0.00 | 1.12 | 0.37 ± 0.02 | 4.91 | 0.25 ± 0.00 | 1.56 | 0.49 ± 0.00 | 0.98 |
C20:5 n3 | 0.24 ± 0.00 | 0.76 | 0.22 ± 0.00 | 1.33 | 0.23 ± 0.02 | 7.01 | 0.18 ± 0.00 | 1.81 | 0.35 ± 0.01 | 1.62 |
C22:5 | 0.22 ± 0.00 | 1.73 | 0.20 ± 0.00 | 2.15 | 0.39 ± 0.02 | 5.44 | 0.23 ± 0.00 | 1.92 | 0.32 ± 0.01 | 1.59 |
C22:6 n3 | 0.07 ± 0.00 | 2.60 | 0.07 ± 0.00 | 4.38 | <0.15 | - | 0.04 ± 0.00 | 9.95 | 0.09 ± 0.00 | 5.06 |
Unreported FA | 2.77 ± 0.12 | 4.34 | 2.26 ± 0.08 | 3.35 | 14.44 ± 0.55 | 3.81 | 2.52 ± 0.07 | 2.85 | 1.86 ± 0.05 | 2.45 |
SUM FA | 26.97 ± 0.37 | 1.36 | 23.81 ± 0.20 | 0.82 | 182.37 ± 4.95 | 2.71 | 31.86 ± 0.39 | 1.21 | 11.03 ± 0.20 | 1.77 |
Variables | Levels Used | Optimum Conditions | ||||
---|---|---|---|---|---|---|
Temperature (°C) | 60 | 70 | 80 | 100 | 70 | |
Time (hr) | 1 | 2 | 3 | 2 | ||
Concentration (% H2SO4 in methanol) | 3 | 5 | 7 | 9 | 5 | |
Acid and toluene volume (mL) | 2 | 3 | 4 | 5 | 6 | 4 (each) |
Sample weight (mg) | 100 | 200 | 300 | 400 | 500 | 300 |
Moisture (%) | 0 | 2.5 | 5 | 7.5 | 0–7.5% |
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Agnew, M.P.; Craigie, C.R.; Weralupitiya, G.; Reis, M.M.; Johnson, P.L.; Reis, M.G. Comprehensive Evaluation of Parameters Affecting One-Step Method for Quantitative Analysis of Fatty Acids in Meat. Metabolites 2019, 9, 189. https://doi.org/10.3390/metabo9090189
Agnew MP, Craigie CR, Weralupitiya G, Reis MM, Johnson PL, Reis MG. Comprehensive Evaluation of Parameters Affecting One-Step Method for Quantitative Analysis of Fatty Acids in Meat. Metabolites. 2019; 9(9):189. https://doi.org/10.3390/metabo9090189
Chicago/Turabian StyleAgnew, Michael P., Cameron R. Craigie, Gayani Weralupitiya, Marlon M. Reis, Patricia L. Johnson, and Mariza G. Reis. 2019. "Comprehensive Evaluation of Parameters Affecting One-Step Method for Quantitative Analysis of Fatty Acids in Meat" Metabolites 9, no. 9: 189. https://doi.org/10.3390/metabo9090189