
Very Long Chain Fatty Acids, Pristanic and Phytanic Acids in Serum GC-MS Analysis Kit
Peroxisomes are organelles surrounded by membrane in the form of spheres. Peroxisomes play an important role in beta oxidation of very long chain fatty acids (C22:0, C24:0, C26:0) and alpha oxidation of 3-methyl fatty acids (phytanic acid). The Zellweger syndrome is best known peroxisomal disorders. Zellweger syndrome is a rare disease characterized by decreased or absent peroxisomes in the cells of the liver, kidney and brain. Increased levels of very long chain fatty acids (VLCFA) in blood and decreased plasmalogen levels in erythrocytes are the most important parameters to aid in diagnosis.
Very long-chain fatty acids, (VLCFA) pristanic and phytanic acid analysis to aid in diagnosis of peroxisomal biogenesis disorders (Zellweger Syndrome Spectrum), X-linked adrenoleukodystrophy (X- ALD). The diagnosis of X-ALD may be raised by the above clinical signs or symptoms, including isolated adrenal insufficiency and is commonly achieved by laboratory evaluation of increased concentrations of VLCFA in plasma. Testing typically includes eight VLCFA parameters: the level of C22:0, C24:0, C26:0, pristanic and phytanic acid and the ratio of C26:0/ C22:0, C24:0/C22:0, pristanic acid/phytanic acid.
Highlights
All analytes, including pristanic phytanic acid, are detected in a single sample preparation.
Consuming small volume of patient’s sample.
Long life span of GC column
Parameters
Matrix
Sample Preparation
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Step 1Take 200 μl serum sample into a glass vial
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Step 2Reagent mix prepare into the volumetric flask on ice in fume hood. Reagent 3 is added drop-wise while swirling on R1 and R2, vortex for 10 seconds
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Step 3600 μl of Reagent-mix, vortex for 15 seconds.
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Step 4Put the vial into the empty glass tube. İncubate in water bath at 90 min 70°C.
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Step 5After cooling to room temperature, the methyl derivatives are transferred in eppendorf, extracted 800 µl Reagent 4 and centrifuge at 3500rpm 2 min.
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Step 6Pipet 600 µl extracted serum samples are dried under stream of nitrogen, dissolved 150 µl Reagent 4, and vortex 10 sec.
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Step 7Transfer the final solution into the insert of HPLC vials.

Chromatogram
METHOD PERFORMANCE
Compound | R2 7890-MSD | LOQ nmol/ml 7890-MSD | Recovery % 7890-MSD | |
Repeatability %RSD 7890-MSD | ||||
Pristanic Acid | 0,9991 | 0,18 | 100,615 | 1,365 |
Phytanic Acid | 0,9999 | 1,635 | 93,522 | 1,919 |
Behenic Acid (C22:0) | 0,9946 | 3,046 | 83,163 | 1,087 |
Lignoceric Acid (C24:0) | 0,9949 | 2,161 | 99,706 | 1,406 |
Cerotic Acid (C26:0) | 0,9933 | 0,159 | 119,49 | 3,031 |