Targeted Metabolomic Profiling of Total Fatty Acids in Human Plasma by Liquid Chromatography-Tandem Mass Spectrometry
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals and Solvents
2.2. Standard Solutions
2.3. Study Samples
2.4. DAABD-AE Derivatization Reaction Optimization
2.5. Sample Preparation
2.6. LC-MS/MS System and Operating Conditions
2.7. Method Validation
3. Results and Discussion
3.1. Derivatization of FAs with DAABD-AE
3.2. Method Development
3.2.1. Derivatization of FAs with DAABD-AE
3.2.2. Chromatographic Separation
3.2.3. Linearity, LOD, and LOQ
3.2.4. Imprecision and Recovery
3.3. Determination of FAs Reference Intervals
3.4. Diagnostic Application on Samples from Patients with Inborn Errors of Metabolism
3.5. Method Comparison
4. Conclusions
Author Contributions
Funding
Conflicts of Interest
Compliance with Ethics Guidelines
References
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Compound | Retention Time (min) | IS * | Precursor Ion (m/z) | Linear Range (μmol/L) | LOD # | LOQ § |
---|---|---|---|---|---|---|
Octanoic acid, C8:0 | 1.6 | (A) | 455.3 | 0.75–75 | 9.9 | 33.0 |
Decenoic acid, C10:1 | 2.0 | (B) | 481.3 | - | ||
Decanoic acid, C10:0 | 2.5 | (B) | 483.3 | 3.75–375 | 14.0 | 46.7 |
Lauroleic acid, C12:1 | 2.8 | (C) | 509.2 | - | ||
Lauric acid, C12:0 | 3.2 | (C) | 511.2 | 7.5–750 | 12.7 | 42.3 |
Tetradecadienoic acid, C14:2 | 3.0 | (D) | 535.4 | - | ||
Myristoleic acid, C14:1 | 3.4 | (D) | 537.4 | - | ||
Myristic acid, C14:0 | 3.8 | (D) | 539.4 | 7.5–750 | 9.0 | 30.0 |
Hexadecenoic acid, C16:1w9 | 3.9 | (E) | 565.4 | 7.5–750 | 8.6 | 28.7 |
Palmitic acid, C16:0 | 4.7 | (E) | 567.3 | 60–6000 | 4.2 | 14.0 |
Stearidonic acid, C18:4 | 3.5 | (F) | 587.4 | - | ||
α-Linolenic acid, C18:3w3 | 3.8 | (F) | 589.4 | 3.75–375 | 5.1 | 17.0 |
Linoleic acid, C18:2w6 | 4.2 | (F) | 591.4 | - | ||
Oleic acid, C18:1w9 | 4.9 | (F) | 593.4 | - | 5.9 | 19.7 |
Stearic acid, C18:0 | 5.5 | (F) | 595.4 | 30–3000 | 6.8 | 22.7 |
EPA, C20:5w3 | 3.8 | (H) | 613.4 | 1.9–188 | 14.0 | 46.7 |
Arachidonic acid, C20:4w6 | 4.2 | (H) | 615.4 | 22.5–2250 | 8.6 | 28.7 |
h-γ-Linolenic acid, C20:3w6 | 4.5 | (H) | 617.4 | - | ||
Eicosadienoic acid, C20:2 | 5.1 | (H) | 619.4 | - | ||
Gondoic acid, C20:1 | 5.6 | (H) | 621.4 | - | ||
Arachidic acid, C20:0 | 6.1 | (H) | 623.4 | 0.75–75 | 9.0 | 30.0 |
Pristanic acid, C19:0 branched | 5.3 | (G) | 609.2 | 0.75–75 | 11.4 | 38.0 |
Phytanic acid, C20:0 branched | 5.7 | (H) | 623.2 | 0.75–75 | 9.6 | 32.0 |
DHA, C22:6w3 | 4.1 | (I) | 639.2 | - | ||
DPA, C22:5w3 | 4.6 | (I) | 641.2 | 0.75–75 | 14.3 | 47.7 |
DTA, C22:4w6 | 4.9 | (I) | 643.2 | - | ||
Docosatrienoic acid, C22:3 | 5.5 | (I) | 645.2 | - | ||
Docosadienoic acid, C22:2 | 5.8 | (I) | 647.2 | - | ||
Docosenoic acid, C22:1 | 6.2 | (I) | 649.2 | - | ||
Docosanoic acid, C22:0 | 6.6 | (I) | 651.2 | 3.75–375 | 10.3 | 34.3 |
Nervonic acid, C24:1 | 6.6 | (J) | 677.3 | - | ||
Tetracosanoic acid, C24:0 | 7.1 | (J) | 679.3 | 3.75–375 | 9.8 | 32.7 |
Hexacosenoic acid, C26:1 | 7.1 | (K) | 705.3 | - | ||
Hexacosanoic acid, C26:0 | 7.9 | (K) | 707.3 | 0.15–15 | 15.1 | 50.3 |
Octacosenoic acid, C28:1 | 7.9 | (K) | 733.3 | - | ||
Montanic acid, C28:0 | 9.1 | (K) | 735.3 | - |
Compound | Sample | Concentration (μ mol/L) | Intra-Day (n = 15) | Inter-Day (n = 20) | Recovery (%) | ||||
---|---|---|---|---|---|---|---|---|---|
Mean | SD | CV (%) | Mean | SD | CV (%) | ||||
C8:0 | QC 1 | 38.0 | 36.6 | 2.4 | 6.6 | 36.3 | 0.6 | 1.8 | 98.7 |
QC 2 | 72.4 | 73.4 | 2.5 | 3.4 | 73.4 | 2.5 | 3.4 | ||
C10:0 | QC 1 | 86.2 | 88.2 | 3.9 | 4.4 | 87.3 | 2.0 | 2.3 | 101.0 |
QC 2 | 282.4 | 281.6 | 9.1 | 3.2 | 284.3 | 3.5 | 1.2 | ||
C12:0 | QC 1 | 209.7 | 201.7 | 7.9 | 3.9 | 200.9 | 3.7 | 1.8 | 98.5 |
QC 2 | 580.5 | 588.7 | 11.7 | 2.0 | 584.9 | 5.3 | 0.9 | ||
C14:0 | QC 1 | 315.2 | 308.2 | 21.6 | 7.0 | 314.6 | 3.7 | 1.2 | 99.2 |
QC 2 | 693.1 | 685.6 | 8.0 | 1.2 | 695.7 | 7.1 | 1.0 | ||
C16:1 | QC 1 | 406.6 | 397.8 | 25.0 | 6.3 | 398.5 | 3.9 | 1.0 | 98.8 |
QC 2 | 675.7 | 671.9 | 28.8 | 4.3 | 675.7 | 8.8 | 1.3 | ||
C16:0 | QC 1 | 3663.1 | 3610.3 | 367.6 | 10.2 | 3697.7 | 46.0 | 1.2 | 99.5 |
QC 2 | 5538.9 | 5620.3 | 213.7 | 3.8 | 5367.8 | 534.3 | 10.0 | ||
C18:3w3 | QC 1 | 124.8 | 129.9 | 7.5 | 5.8 | 127.6 | 8.5 | 6.7 | 100.8 |
QC 2 | 330.8 | 325.6 | 21.9 | 6.7 | 324.8 | 3.8 | 1.2 | ||
C18:0 | QC 1 | 1655.4 | 1639.7 | 113.3 | 6.9 | 1640.6 | 35.1 | 2.1 | 99.4 |
QC 2 | 2902.7 | 2889.7 | 39.8 | 1.4 | 2895.6 | 63.8 | 2.2 | ||
C20:5w3 | QC 1 | 94.9 | 94.6 | 2.5 | 2.6 | 95.8 | 2.5 | 2.6 | 101.1 |
QC 2 | 177.2 | 181.7 | 6.5 | 3.6 | 179.7 | 2.4 | 1.3 | ||
C20:4 | QC 1 | 1340.2 | 1546.2 | 67.3 | 4.4 | 1621.3 | 145.4 | 9.0 | 106.4 |
QC 2 | 2214.1 | 1965.4 | 41.8 | 2.1 | 2224.7 | 36.7 | 1.7 | ||
C20:0 | QC 1 | 32.5 | 33.5 | 0.4 | 1.3 | 31.2 | 1.3 | 4.3 | 94.5 |
QC 2 | 75.4 | 67.8 | 2.0 | 2.9 | 67.0 | 2.5 | 3.7 | ||
Pristanic | QC 1 | 16.6 | 16.9 | 0.6 | 3.4 | 16.4 | 1.0 | 6.2 | 98.5 |
QC 2 | 55.9 | 55.1 | 2.3 | 4.2 | 53.4 | 2.0 | 3.8 | ||
Phytanic | QC 1 | 16.8 | 17.0 | 1.0 | 5.7 | 16.3 | 0.4 | 2.3 | 98.0 |
QC 2 | 56.2 | 55.0 | 2.0 | 3.6 | 54.1 | 1.5 | 2.8 | ||
C22:5w6 | QC 1 | 42.6 | 42.9 | 3.9 | 9.1 | 43.3 | 1.2 | 2.7 | 101.5 |
QC 2 | 70.2 | 72.0 | 3.2 | 4.4 | 71.2 | 1.9 | 2.7 | ||
C22:0 | QC 1 | 124.9 | 124.0 | 2.0 | 1.6 | 123.3 | 1.4 | 1.2 | 99.2 |
QC 2 | 317.7 | 317.5 | 12.6 | 4.0 | 314.4 | 5.1 | 1.6 | ||
C24.0 | QC 1 | 126.0 | 127.6 | 2.6 | 2.1 | 124.2 | 1.6 | 1.3 | 100.8 |
QC 2 | 309.1 | 315.8 | 7.2 | 2.3 | 313.4 | 2.3 | 0.7 | ||
C26.0 | QC 1 | 3.8 | 3.7 | 0.1 | 3.0 | 3.7 | 0.1 | 1.5 | 97.8 |
QC 2 | 11.8 | 11.4 | 0.2 | 1.9 | 11.7 | 0.2 | 1.5 |
Compound | <1 Month | 1–12 Month | 1–18 Year | >18 Year | ||||
---|---|---|---|---|---|---|---|---|
Low | High | Low | High | Low | High | Low | High | |
Octanoic acid, C8:0 | 22 | 53 | 21 | 60 | 22 | 62 | 18 | 41 |
Decenoic acid, C10:1 | 0.2 | 1.7 | 0.2 | 2.3 | 0.1 | 1.3 | 0.1 | 1.1 |
Decanoic acid, C10:0 | 12 | 46 | 9 | 60 | 10 | 57 | 9 | 41 |
Lauroleic acid, C12:1 | 0.1 | 4.0 | 0.2 | 2.8 | 0.2 | 1.6 | 0.2 | 2.4 |
Lauric acid, C12:0 | 21 | 165 | 19 | 211 | 25 | 202 | 35 | 152 |
Tetradecadienoic acid, C14:2 | 0.1 | 4.1 | 0.2 | 9.4 | 0.2 | 4.3 | 0.1 | 3.6 |
Myristoleic acid, C14:1 | 0.7 | 6.8 | 0.8 | 10.1 | 1.0 | 20.5 | 1.2 | 14.2 |
Myristic acid, C14:0 | 35 | 367 | 43 | 327 | 37 | 293 | 40 | 337 |
Hexadecenoic acid, C16:1w9 | 163 | 654 | 74 | 517 | 49 | 590 | 72 | 514 |
Palmitic acid, C16:0 | 1304 | 3654 | 1289 | 3595 | 554 | 3411 | 1238 | 3999 |
Stearidonic acid, C18:4 | 0.0 | 21.1 | 0.1 | 17.5 | 0.2 | 27.2 | 0 | 29 |
α-Linolenic acid, C18:3w3 | 0.5 | 46.9 | 1.6 | 66.8 | 5.7 | 62.7 | 3 | 44 |
Linoleic acid, C18:2w6 | 216 | 1750 | 620 | 2544 | 655 | 2193 | 672 | 2961 |
Oleic acid, C18:1w9 | 925 | 3250 | 1237 | 4943 | 857 | 4041 | 816 | 4433 |
Stearic acid, C18:0 | 562 | 1410 | 580 | 1553 | 253 | 1414 | 511 | 1507 |
EPA, C20:5w3 | 10 | 118 | 7 | 73 | 9 | 92 | 6 | 88 |
Arachidonic acid, C20:4w6 | 622 | 1652 | 303 | 1316 | 122 | 1155 | 275 | 1576 |
h-γ-Linolenic acid, C20:3w6 | 30 | 111 | 17 | 113 | 29 | 149 | 23 | 131 |
Eicosadienoic acid, C20:2 | 4 | 33 | 5 | 38 | 5 | 23 | 5 | 22 |
Gondoic acid, C20:1 | 6 | 36 | 8 | 49 | 5 | 38 | 6 | 35 |
Arachidic acid, C20:0 | 11 | 37 | 5 | 40 | 5 | 28 | 5 | 33 |
Pristanic acid, C19:0 branched | 1.1 | 3.0 | 1.1 | 2.8 | 1.2 | 3.0 | 1.3 | 3.0 |
Phytanic acid, C20:0 branched | 1.6 | 3.6 | 1.8 | 4.9 | 1.7 | 10.3 | 1.8 | 8.0 |
DHA, C22:6w3 | 9 | 60 | 15 | 64 | 5 | 45 | 4 | 39 |
DPA, C22:5w3 | 13 | 88 | 7 | 73 | 8 | 52 | 4 | 43 |
DTA, C22:4w6 | 19 | 68 | 14 | 64 | 14 | 53 | 9 | 61 |
Docosatrienoic acid, C22:3 | 1 | 7 | 0 | 5 | 1 | 6 | 0 | 4 |
Docosadienoic acid, C22:2 | 3 | 15 | 3 | 14 | 2 | 11 | 2 | 10 |
Docosenoic acid, C22:1 | 15 | 45 | 14 | 35 | 12 | 38 | 11 | 39 |
Docosanoic acid, C22:0 | 27 | 60 | 21 | 100 | 28 | 70 | 28 | 77 |
Nervonic acid, C24:1 | 77 | 257 | 82 | 220 | 54 | 210 | 68 | 267 |
Tetracosanoic acid, C24:0 | 18 | 59 | 17 | 70 | 21 | 65 | 17 | 76 |
Hexacosenoic acid, C26:1 | 0.9 | 3.2 | 0.3 | 2.0 | 0.2 | 1.9 | 0.2 | 1.9 |
Hexacosanoic acid, C26:0 | 0.3 | 1.0 | 0.2 | 1.0 | 0.2 | 1.2 | 0.3 | 1.1 |
Octacosenoic acid, C28:1 | 0.01 | 0.10 | 0.00 | 0.16 | 0.00 | 0.10 | 0.01 | 0.09 |
Montanic acid, C28:0 | 0.07 | 0.19 | 0.04 | 0.18 | 0.03 | 0.20 | 0.03 | 0.15 |
Total fatty acids (mmol/L) | 4.1 | 14.1 | 4.4 | 16.2 | 2.8 | 14.4 | 3.9 | 16.6 |
Total saturated fatty acids (mmol/L) | 2.0 | 5.9 | 2.0 | 6.0 | 1.0 | 5.6 | 1.9 | 6.3 |
Total MUFA (mmol/L) | 1.2 | 4.3 | 1.4 | 5.8 | 1.0 | 4.9 | 1.0 | 5.3 |
Total PUFA (mmol/L) | 0.9 | 4.0 | 1.0 | 4.4 | 0.9 | 3.9 | 1.0 | 5.0 |
Patient | 1 | 2 | 3 | 4 | 5 | Reference Interval |
---|---|---|---|---|---|---|
Age | 10 day | 10.8 year | 2.6 year | 22.2 year | 2 year | |
Sex | F | M | F | M | F | |
Diagnosis | Severe PBD | X-ALD | RD | AMN | Mild PBD | |
PRA | 1.6 | 1.9 | 3.7 | 1.7 | 4.2 | 1.2–3.0 |
PHA | 4.3 | 4.3 | 23.2 | 4.7 | 15.2 | 1.7–10.3 |
C22:0 | 46.9 | 40.1 | 47.0 | 52.3 | 25.1 | 21–100 |
C24:0 | 63.8 | 57.0 | 28.0 | 86.7 | 24.5 | 17–76 |
C26:0 | 17.4 | 2.2 | 0.9 | 3.0 | 2.1 | 0.2–1.2 |
C28:0 | 1.82 | 0.38 | 0.19 | 0.38 | 0.20 | 0.03–0.2 |
C24:0/C22:0 | 1.36 | 1.42 | 0.76 | 1.66 | 0.98 | ≤1.20 |
C26:0/C22:0 | 0.372 | 0.054 | 0.019 | 0.057 | 0.082 | ≤0.022 |
C28:0/C22:0 | 0.0388 | 0.0094 | 0.0041 | 0.0073 | 0.0080 | ≤0.0045 |
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Aidaros, A.A.; Sharma, C.; Langhans, C.-D.; G. Okun, J.; Hoffmann, G.F.; Dasouki, M.; Chakraborty, P.; Aljasmi, F.; Y. Al-Dirbashi, O. Targeted Metabolomic Profiling of Total Fatty Acids in Human Plasma by Liquid Chromatography-Tandem Mass Spectrometry. Metabolites 2020, 10, 400. https://doi.org/10.3390/metabo10100400
Aidaros AA, Sharma C, Langhans C-D, G. Okun J, Hoffmann GF, Dasouki M, Chakraborty P, Aljasmi F, Y. Al-Dirbashi O. Targeted Metabolomic Profiling of Total Fatty Acids in Human Plasma by Liquid Chromatography-Tandem Mass Spectrometry. Metabolites. 2020; 10(10):400. https://doi.org/10.3390/metabo10100400
Chicago/Turabian StyleAidaros, Anas Al, Charu Sharma, Claus-Dieter Langhans, Jürgen G. Okun, Georg F. Hoffmann, Majed Dasouki, Pranesh Chakraborty, Fatma Aljasmi, and Osama Y. Al-Dirbashi. 2020. "Targeted Metabolomic Profiling of Total Fatty Acids in Human Plasma by Liquid Chromatography-Tandem Mass Spectrometry" Metabolites 10, no. 10: 400. https://doi.org/10.3390/metabo10100400