Mass spectrometric determination of fatty acids

1. A method for determining an amount of one or more very long chain fatty acids (VLCFA) and/or branched chain fatty acids (BCFA) in a sample by mass spectrometry, the method comprising:

(a) subjecting the sample to an ionization source operating in positive ionization mode to generate one or more VLCFA and/or BCFA ions detectable by mass spectrometry;

(b) determining the amount of the one or more VLCFA and/or BCFA ions by mass spectrometry; and

(c) relating the amount of the one or more VLCFA and/or BCFA ions determined in step (b) to the amount of the VLCFA and/or BCFA in the sample;

wherein the VLCFA and/or BCFA are underivatized prior to ionization.

2. The method of claim 1 , wherein said one or more VLCFA are selected from the group consisting of docosanoic acid, tetracosanoic acid, and hexacosanoic acid.

3. The method of claim 2 , wherein said one or more VLCFA ions comprise ions with a mass to charge ratio (m/z) of 339.3±0.5.

4. The method of claim 2 , wherein said one or more VLCFA ions comprise ions with a mass to charge ratio (m/z) of 367.3±0.5.

5. The method of claim 2 , wherein said one or more VLCFA ions comprise ions with a mass to charge ratio (m/z) of 395.4±0.5.

6. The method of claim 1 , wherein said one or more BCFA are selected from the group consisting of pristanic acid and phytanic acid.

7. The method of claim 6 , wherein said one or more BCFA ions comprise ions with a mass to charge ratio (m/z) of 297.3±0.5.

8. The method of claim 6 , wherein said one or more BCFA ions comprise ions with a mass to charge ratio (m/z) of 311.2±0.5.

9. The method of claim 1 , wherein the ionization source is electron ionization, chemical ionization, electrospray ionization (ESI), photon ionization, photoionization, atmospheric pressure photoionization (APPI), laser diode thermal desorption (LDTD), fast atom bombardment (FAB), liquid secondary ionization (LSI), matrix assisted laser desorption ionization (MALDI), field ionization, field desorption, thermospray ionization, plasmaspray ionization, surface enhanced laser desorption ionization (SELDI), inductively coupled plasma (ICP), or particle beam ionization.

10. The method of claim 1 , wherein the amounts of one or more VLCFA and one or more BCFA are separately determined.

11. The method of claim 10 , wherein said one or more VLCFA are selected from the group consisting of docosanoic acid, tetracosanoic acid, and hexacosanoic acid, and wherein said one or more BCFA are selected from the group consisting of pristanic acid and phytanic acid.

12. The method of claim 1 , wherein said sample comprises a human sample comprising plasma or serum.

13. The method of claim 1 , wherein said sample is subjected to a hydrolyzing agent prior to ionization.

14. The method of claim 1 , wherein the sample is subjected to liquid/liquid extraction prior to ionization.

15. The method of claim 1 , wherein the one or more VLCFA and/or BCFA are subjected to a liquid chromatography column prior to ionization.

16. The method of claim 15 , wherein the liquid chromatography column comprises a high performance liquid chromatography (HPLC), reverse phase liquid chromatography (RPLC), turbulent flow liquid chromatography (TFLC), or high turbulence liquid chromatography (HTLC).

17. The method of claim 1 , wherein the method further comprises determining the amount of the one or more internal standards.

18. The method of claim 17 , wherein the internal standard is pristanic acid- 2 H 3 phytanic acid- 2 H 3 , docosanoic acid- 2 H 3 , tetracosanoic acid- 2 H 3 , or hexacosanoic acid- 2 H 3 .

19. A method of diagnosing or monitoring a peroxisomal disorder comprising determining an amount of one or more very long chain fatty acids (VLCFA) and/or branched chain fatty acids (BCFA) in a patient sample by steps of claim 1 .

20. The method of claim 19 , wherein an abnormal level of VLCFA and/or BCFA is indicative of the peroxisomal disorder.

21. The method of claim 19 , wherein the peroxisomal disorder is Zellweger syndrome, pseudo-Zellweger syndrome, infantile and adult Refsum disease, adrenoleukodystrophy, rhizomelic chondrodysplasia punctata type 1 (RCDP-1), D-bifunctional protein deficiency, or acyl-coA oxidase deficiency.

22. A method for determining an amount of one or more very long chain fatty acids (VLCFA) and/or branched chain fatty acids (BCFA) in a sample by mass spectrometry, the method comprising:

(a) subjecting the sample to a hexane extraction;

(b) subjecting the sample to liquid chromatography;

(c) subjecting the sample to an ionization source operating in positive ionization mode to generate one or more VLCFA and/or BCFA ions detectable by mass spectrometry;

(d) determining the amount of the one or more VLCFA and/or BCFA ions by mass spectrometry; and

(e) relating the amount of the one or more VLCFA and/or BCFA ions determined in step (d) to the amount of the VLCFA and/or BCFA in the sample.

23. The method of claim 22 , wherein said one or more VLCFA are selected from the group consisting of docosanoic acid, tetracosanoic acid, and hexacosanoic acid.

24. The method of claim 22 , wherein said one or more BCFA are selected from the group consisting of pristanic acid and phytanic acid.

25. The method of claim 22 , wherein the amounts of one or more VLCFA and one or more BCFA are separately determined.

26. The method of claim 22 , wherein the ionization source is electron ionization, chemical ionization, electrospray ionization (ESI), photon ionization, photoionization, atmospheric pressure photoionization (APPI), laser diode thermal desorption (LDTD), fast atom bombardment (FAB), liquid secondary ionization (LSI), matrix assisted laser desorption ionization (MALDI), field ionization, field desorption, thermospray ionization, plasmaspray ionization, surface enhanced laser desorption ionization (SELDI), inductively coupled plasma (ICP), or particle beam ionization.

27. The method of claim 22 , wherein said sample comprises a human sample comprising plasma or serum.

28. The method of claim 22 , wherein said sample is subjected to an acid hydrolysis prior to ionization.

29. The method of claim 22 , wherein the method further comprises determining the amount of the one or more internal standards.

30. The method of claim 29 , wherein the internal standard is pristanic acid- 2 H 3 phytanic acid- 2 H 3 , docosanoic acid- 2 H 3 , tetracosanoic acid- 2 H 3 , or hexacosanoic acid- 2 H 3 .

31. A method of diagnosing or monitoring a peroxisomal disorder comprising determining an amount of one or more very long chain fatty acids (VLCFA) and/or branched chain fatty acids (BCFA) in a patient sample by steps of claim 22 .

32. The method of claim 31 , wherein an abnormal level of VLCFA and/or BCFA is indicative of the peroxisomal disorder.

33. The method of claim 31 , wherein the peroxisomal disorder is Zellweger syndrome, pseudo-Zellweger syndrome, infantile and adult Refsum disease, adrenoleukodystrophy, rhizomelic chondrodysplasia punctata type 1 (RCDP-1), D-bifunctional protein deficiency, or acyl-coA oxidase deficiency.