Optimization of Ultrasonic-assisted Extraction of Fatty Acids in Seeds of Brucea Javanica (L.) Merr. from Different Sources and Simultaneous Analysis Using High-Performance Liquid Chromatography with Charged Aerosol Detection
Abstract
:1. Introduction
2. Results and Discussion
2.1. Validation of HPLC-CAD Method
2.1.1. Calibration Curves, Limits of Detection, and Quantification
2.1.2. Precision
2.1.3. Accuracy
2.2. Effect of Independent Variables on the Content of FAs and the Oil Yield
2.3. Model Fitting
2.4. Analysis of Response Surface
2.5. Optimization of Extraction Parameters and Validation of the Model
2.6. Effect of Extraction Times on Content of FAs and Oil Yield
2.7. Comparison of Content of FAs
2.8. HPLC Fingerprint, Cluster, and Principal Component Analysis
3. Materials and Methods
3.1. Materials and Standards
3.2. Extraction of Oil
3.3. Preparation of Sample Solution
3.4. HPLC-CAD Analysis
3.5. Validation of HPLC-CAD Method
3.5.1. Calibration Curves, Limits of Detection, and Quantification
3.5.2. Precision
3.5.3. Accuracy
3.6. Content of FAs and Oil Yield Determination
3.7. Optimization of UAE
3.8. Determination of Extraction Times
3.9. Data Analysis
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of interest
Abbreviations
ANOVA | Analysis of variance |
BBD | Box-Behnken design |
BJO | Brucea javanica (L.) Merr. seed oil |
CAD | Charged aerosol detector |
FAs | Fatty acids |
RSM | Response surface methodology |
TCM | Traditional Chinese Medicine |
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Sample Availability: Not available. |
Compound | Retention Time (min) | Linear Range (mg/mL) | Equation | R2 | LOD (μg/mL) | LOQ (μg/mL) |
---|---|---|---|---|---|---|
Linolenic acid | 6.95 | 0.0012–0.0349 | y = 63.533x + 0.0164 | 0.9999 | 0.504 | 1.080 |
Linoleic acid | 9.95 | 0.0109–0.5452 | y = 44.152x + 0.8404 | 0.9983 | 0.368 | 2.624 |
Palmitic acid | 14.45 | 0.0084–0.4204 | y = 53.760x + 0.2189 | 0.9996 | 1.400 | 4.032 |
Oleic acid | 15.56 | 0.0161–0.8032 | y = 46.574x + 1.3419 | 0.9983 | 1.376 | 3.856 |
Stearic acid | 26.65 | 0.0068–0.2030 | y = 81.835x + 0.0361 | 0.9996 | 1.958 | 6.800 |
Compound | Concentration (mg/mL) | Intra-day (n = 6) | Inter-day (n = 3) |
---|---|---|---|
RSD (%) | RSD (%) | ||
Linolenic acid | 0.0012 | 1.80 | 1.19 |
0.0116 | 1.07 | 1.36 | |
0.0349 | 1.71 | 1.99 | |
Linoleic acid | 0.0112 | 1.82 | 2.17 |
0.1118 | 0.37 | 1.42 | |
0.3353 | 1.04 | 2.29 | |
Palmitic acid | 0.0088 | 1.33 | 1.21 |
0.0877 | 0.48 | 1.94 | |
0.2630 | 1.36 | 2.02 | |
Oleic acid | 0.0231 | 1.97 | 2.06 |
0.2310 | 0.26 | 1.44 | |
0.6931 | 1.00 | 2.05 | |
Stearic acid | 0.0066 | 1.48 | 2.71 |
0.0664 | 0.44 | 2.86 | |
0.1992 | 0.98 | 2.64 |
Compound | Initial(mg) | Amount(mg) | Add(mg) | Found ± SD(mg) | Recovery (%) | RSD (%, n = 3) |
---|---|---|---|---|---|---|
Linolenic acid | 8.87 | 0.0177 | 0.0089 | 0.0265 ± 0.0006 | 98.30 | 2.79 |
8.96 | 0.0179 | 0.0090 | 0.0272 ± 0.0002 | 103.68 | ||
8.71 | 0.0174 | 0.0087 | 0.0264 ± 0.0003 | 102.55 | ||
8.48 | 0.0170 | 0.0170 | 0.0331 ± 0.0005 | 95.19 | 2.17 | |
8.71 | 0.0174 | 0.0174 | 0.0346 ± 0.0005 | 98.65 | ||
8.63 | 0.0173 | 0.0173 | 0.0336 ± 0.0006 | 94.91 | ||
8.11 | 0.0162 | 0.0243 | 0.0398 ± 0.0006 | 96.71 | 2.20 | |
8.04 | 0.0161 | 0.0241 | 0.0404 ± 0.0004 | 101.03 | ||
8.23 | 0.0165 | 0.0247 | 0.0408 ± 0.0004 | 98.46 | ||
Linoleic acid | 8.87 | 1.2306 | 0.6153 | 1.8451 ± 0.0116 | 99.87 | 2.66 |
8.96 | 1.2431 | 0.6216 | 1.8414 ± 0.0037 | 96.25 | ||
8.71 | 1.2084 | 0.6042 | 1.7817 ± 0.0133 | 94.88 | ||
8.48 | 1.1765 | 1.1765 | 2.3642 ± 0.0061 | 100.95 | 2.21 | |
8.71 | 1.2084 | 1.2084 | 2.4752 ± 0.0070 | 104.83 | ||
8.63 | 1.1973 | 1.1973 | 2.4545 ± 0.0041 | 105.00 | ||
8.11 | 11252 | 1.6878 | 2.8308 ± 0.0307 | 101.06 | 1.84 | |
8.04 | 1.1155 | 1.6732 | 2.8386 ± 0.0064 | 102.98 | ||
8.23 | 1.1418 | 1.7127 | 2.9375 ± 0.1431 | 104.84 | ||
Palmitic acid | 8.87 | 0.6905 | 0.3453 | 1.0414 ± 0.0146 | 101.63 | 1.49 |
8.96 | 0.6975 | 0.3478 | 1.0604 ± 0.0014 | 104.03 | ||
8.71 | 0.6781 | 0.3390 | 1.0212 ± 0.0005 | 101.21 | ||
8.48 | 0.6602 | 0.6602 | 1.2946 ± 0.0044 | 96.10 | 1.82 | |
8.71 | 0.6781 | 0.6781 | 1.3456 ± 0.0027 | 98.44 | ||
8.63 | 0.6718 | 0.6718 | 1.3411 ± 0.0080 | 99.62 | ||
8.11 | 0.6314 | 0.9470 | 1.6031 ± 0.0235 | 102.60 | 0.94 | |
8.04 | 0.6259 | 0.9389 | 1.6065 ± 0.0033 | 104.44 | ||
8.23 | 0.6407 | 0.9611 | 1.6301 ±0.0197 | 102.94 | ||
Oleic acid | 8.87 | 2.9146 | 1.4573 | 4.3238 ± 0.0055 | 96.70 | 2.71 |
8.96 | 2,9442 | 1.4721 | 4.4206 ± 0.0068 | 100.29 | ||
8.71 | 2.8620 | 1.4310 | 4.3215 ± 0.0102 | 101.99 | ||
8.48 | 2.7864 | 2.7864 | 5.4940 ± 0.0036 | 97.17 | 1.15 | |
8.71 | 2.8620 | 2.8620 | 5.6369 ± 0.0138 | 96.96 | ||
8.63 | 2.8357 | 2.8357 | 5.5337 ± 0.0057 | 95.14 | ||
8.11 | 2.6649 | 3.9973 | 6.6010 ± 0.0695 | 98.47 | 1.29 | |
8.04 | 2.6419 | 3.9628 | 6.6026 ± 0.0303 | 99.95 | ||
8.23 | 2.7043 | 4.0564 | 6.8029 ± 0.3343 | 101.04 | ||
Stearic acid | 8.87 | 0.3869 | 0.1930 | 0.5709 ± 0.0018 | 95.80 | 2.94 |
8.96 | 0.3899 | 0.1950 | 0.5799 ± 0.0020 | 97.42 | ||
8.71 | 0.3791 | 0.1896 | 0.5713 ± 0.0036 | 101.41 | ||
8.48 | 0.3690 | 0.3690 | 0.7205 ± 0.0037 | 95.24 | 2.33 | |
8.71 | 0.3791 | 0.3791 | 0.7566 ± 0.0038 | 99.60 | ||
8.63 | 0.3756 | 0.3756 | 0.7376 ± 0.0109 | 96.38 | ||
8.11 | 0.3529 | 0.5294 | 0.8560 ± 0.0113 | 95.03 | 1.19 | |
8.04 | 0.3499 | 0.5249 | 0.8498 ± 0.0070 | 95.24 | ||
8.23 | 0.3582 | 0.5373 | 0.8799 ± 0.0062 | 97.10 |
Source | Content of FAs (mg/g) | Oil Yield (%) | ||||||||
---|---|---|---|---|---|---|---|---|---|---|
SS | DF | MS | F | P | SS | DF | MS | F | P | |
Model | 32.63 | 9 | 3.63 | 4.98 | 0.0229 | 178.73 | 9 | 19.86 | 13.52 | 0.0012 |
X1 | 1.06 | 1 | 1.06 | 1.46 | 0.2669 | 3.24 | 1 | 3.24 | 2.20 | 0.1812 |
X2 | 5.53 | 1 | 5.53 | 7.60 | 0.0282 | 18.79 | 1 | 18.79 | 12.79 | 0.009 |
X3 | 10.76 | 1 | 10.76 | 14.80 | 0.0063 | 147.32 | 1 | 147.32 | 100.27 | <0.0001 |
X1X2 | 0.89 | 1 | 0.89 | 1.23 | 0.3045 | 3.06 | 1 | 3.06 | 2.08 | 0.192 |
X1X3 | 0.05 | 1 | 0.05 | 0.07 | 0.8039 | 0.39 | 1 | 0.39 | 0.27 | 0.622 |
X2X3 | 0.69 | 1 | 0.69 | 0.95 | 0.3629 | 2.13 | 1 | 2.13 | 1.45 | 0.2675 |
X12 | 4.57 | 1 | 4.57 | 6.29 | 0.0405 | 2.92 | 1 | 2.92 | 1.99 | 0.2012 |
X22 | 0.44 | 1 | 0.44 | 0.60 | 0.4635 | 0.18 | 1 | 0.18 | 0.12 | 0.7378 |
X32 | 9.35 | 1 | 9.35 | 12.86 | 0.0089 | 0.80 | 1 | 0.80 | 0.55 | 0.4837 |
Residual | 5.09 | 7 | 0.73 | 10.28 | 7 | 1.47 | ||||
Lack of Fit | 1.65 | 3 | 0.55 | 0.64 | 0.6278 | 8.06 | 3 | 2.69 | 4.84 | 0.0808 |
Pure Error | 3.44 | 4 | 0.86 | 2.22 | 4 | 0.56 | ||||
Cor Total | 37.72 | 16 | 189.01 | 16 | ||||||
R2 | 0.8650 | 0.9456 |
Number | Origin | Linolenic Acid (mg/g) | Linoleic Acid (mg/g) | Palmitic Acid (mg/g) | Oleic Acid (mg/g) | Stearic Acid (mg/g) | Total Contents of FAs (mg/g) |
---|---|---|---|---|---|---|---|
S1 | Guangdong-1 | 0.005 ± 0.001 a | 4.588 ± 0.132 | 3.242 ± 0.138 | 16.105 ± 0.489 | 1.469 ± 0.066 | 25.409 |
S2 | Guangdong-2 | 0.092 ± 0.004 | 13.254 ± 0.101 | 6.805 ± 0.117 | 34.278 ± 1.213 | 3.817 ± 0.048 | 58.246 |
S3 | Guangdong-3 | 0.178 ± 0.009 | 14.064 ± 0.082 | 6.977 ± 0.209 | 30.492 ± 0.717 | 3.950 ± 0.078 | 55.661 |
S4 | Guangdong-4 | 0.012 ± 0.001 | 21.593 ± 0.156 | 11.584 ± 0.124 | 65.924 ± 2.023 | 7.497 ± 0.129 | 106.610 |
S5 | Guangdong-5 | 0.064 ± 0.003 | 17.259 ± 0.984 | 8.735 ± 0.274 | 49.433 ± 2.123 | 4.169 ± 0.140 | 79.660 |
S6 | Guangdong-6 | 0.058 ± 0.003 | 20.321 ± 0.331 | 9.755 ± 0.227 | 44.498 ± 0.564 | 6.095 ± 0.100 | 80.727 |
S7 | Guangxi-1 | 0.054 ± 0.003 | 25.486 ± 0.902 | 12.803 ± 0.473 | 77.477 ± 3.706 | 6.876 ± 0.164 | 122.696 |
S8 | Guangxi-2 | 0.056 ± 0.001 | 27.270 ± 0.693 | 12.936 ± 0.219 | 69.677 ± 0.772 | 6.885 ± 3.840 | 116.824 |
S9 | Guangxi-3 | 0.280 ± 0.004 | 22.283 ± 0.568 | 11.176 ± 0.209 | 50.379 ± 2.704 | 6.060 ± 0.204 | 90.178 |
S10 | Fujian-1 | 0.070 ± 0.002 | 8.284 ± 0.052 | 4.660 ± 0.028 | 22.655 ± 0.054 | 1.646 ± 0.033 | 37.315 |
S11 | Fujian-2 | 0.165 ± 0.006 | 16.404 ± 0.170 | 8.522 ± 0.136 | 36.787 ± 0.378 | 4.580 ± 0.069 | 66.458 |
S12 | Hebei | 0.056 ± 0.002 | 8.623 ± 0.086 | 4.020 ± 0.011 | 24.894 ± 0.227 | 1.907 ± 0.034 | 39.500 |
S13 | Yunnan | 0.255 ± 0.008 | 9.007 ± 0.078 | 5.297 ± 0.153 | 20.355 ± 0.578 | 2.441 ± 0.081 | 37.355 |
S14 | Hainan | 0.150 ± 0.004 | 11.178 ± 0.010 | 6.398 ± 0.088 | 30.067 ± 0.537 | 2.646 ± 0.076 | 50.439 |
Independent Variable | Units | Symbol | Coded Levels | ||
---|---|---|---|---|---|
−1 | 0 | 1 | |||
Sonication time | min | X1 | 20 | 30 | 40 |
Liquid-solid ratio | mL/g | X2 | 16:1 | 20:1 | 24:1 |
Ethanol concentration | % | X3 | 90 | 95 | 100 |
Run | Factor 1 (X1) | Factor 2 (X2) | Factor 3 (X3) | Response 1 (Y1) | Response 2 (Y2) |
---|---|---|---|---|---|
Sonication Time (min) | Liquid-solid Ratio (mL/g) | Ethanol Concentration (%) | Content of FAs (mg/g) | Oil Yield (%) | |
1 | −1 | 0 | −1 | 8.71 | 5.00 |
2 | 0 | 0 | 0 | 9.81 | 8.33 |
3 | 0 | 1 | 1 | 11.40 | 14.25 |
4 | 1 | 1 | 0 | 14.16 | 14.25 |
5 | 0 | 0 | 0 | 11.73 | 9.17 |
6 | −1 | 1 | 0 | 12.08 | 9.75 |
7 | 0 | −1 | 1 | 9.32 | 10.83 |
8 | 1 | 0 | 1 | 12.05 | 14.17 |
9 | 0 | −1 | −1 | 8.53 | 4.50 |
10 | 0 | 0 | 0 | 11.70 | 10.00 |
11 | 1 | 0 | −1 | 8.81 | 4.17 |
12 | 0 | 1 | −1 | 8.95 | 5.00 |
13 | −1 | 0 | 1 | 11.51 | 13.75 |
14 | 0 | 0 | 0 | 10.31 | 8.13 |
15 | 0 | 0 | 0 | 10.04 | 8.75 |
16 | 1 | −1 | 0 | 11.14 | 8.33 |
17 | −1 | −1 | 0 | 10.95 | 7.33 |
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Wu, Z.; Li, L.; Li, N.; Zhang, T.; Pu, Y.; Zhang, X.; Zhang, Y.; Wang, B. Optimization of Ultrasonic-assisted Extraction of Fatty Acids in Seeds of Brucea Javanica (L.) Merr. from Different Sources and Simultaneous Analysis Using High-Performance Liquid Chromatography with Charged Aerosol Detection. Molecules 2017, 22, 931. https://doi.org/10.3390/molecules22060931
Wu Z, Li L, Li N, Zhang T, Pu Y, Zhang X, Zhang Y, Wang B. Optimization of Ultrasonic-assisted Extraction of Fatty Acids in Seeds of Brucea Javanica (L.) Merr. from Different Sources and Simultaneous Analysis Using High-Performance Liquid Chromatography with Charged Aerosol Detection. Molecules. 2017; 22(6):931. https://doi.org/10.3390/molecules22060931
Chicago/Turabian StyleWu, Zhuona, Ling Li, Ning Li, Tong Zhang, Yiqiong Pu, Xitong Zhang, Yue Zhang, and Bing Wang. 2017. "Optimization of Ultrasonic-assisted Extraction of Fatty Acids in Seeds of Brucea Javanica (L.) Merr. from Different Sources and Simultaneous Analysis Using High-Performance Liquid Chromatography with Charged Aerosol Detection" Molecules 22, no. 6: 931. https://doi.org/10.3390/molecules22060931