Quantitative Organic Acids in Urine
Human urine is a complex biofluid of diverse small polar metabolites such as organic acids. Organic acidurias are inborn errors of metabolism given rise to by defections of inherited enzymes which lead to accumulation excess amounts of normal organic acids in human urine. For this reason qualitative and quantitative analyses of urinary organic acids are prominent diagnostic tools for the interpretation of metabolite profile that reflects inherited metabolic disorders. Conventionally gas chromatography coupled to mass sepectrometry (GC-MS) is used in spite of disadvantages as limititaton to volatile and thermally stable analytes thereby obligating of time consuming derivatization process.
At this point, “Jasem-Organic Acids” method overcomes abovementioned drawbacks by using liquid chromatography tandem mass spectrometry (LC-MS/MS). Jasem method ensures quantification of 54 organic acids by absence of derivatization simplified sample preperation to diluting. Jasem offers two panels achieving sufficient chromatographic separation for crucial isomers by LC-MS/MS.
Sample Preparation for Urine
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Dilute urine sample 10 times using Reagent-1
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Transfer 100 µL of diluted urine sample to a vial
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Add 50 µL of IS mix and 850 µL of Reagent-1 into the sample and inject to LC-MS/MS system.
- Lıst of Organıc Acıds Analysed ın Panel-1 and Panel-2
- Extracted LC-MS/MS Chromatogram of Panel 1 and Panel 2
- Results
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Lıst of Organıc Acıds Analysed ın Panel-1 and Panel-2
PANEL-1 PANEL-2 2-Methylcitrate 2-OH-Glutaric acid 2-OH-Phenylacetic acid 2-OH-3-Methylpentanoic acid 2-OH-Butyric acid 2-OH-Isocaproic acid 2-Oxoadipic acid 2-OH-Isovaleric acid 3-Methylglutaconic acid 3-OH-Glutaric acid 3-OH-2-Methylbutanoic acid 3-OH-Propanoic acid 3-OH-3-Methylglutaric acid 3-Methylcrotonyl glycine 3-OH-Butyric acid 3-Methylglutaric acid 3-OH-Isobutyrate 3-Methyl-2-oxovaleric acid 3-Phenyllactic acid 3-OH-Isovaleric acid 4-OH-Phenylacetic acid 3-OH-Pentanoic acid Fumaric acid 4-Methyl-2-oxovaleric acid Glycolic acid 4-OH-Phenylpyruvic acid Hexanoylglycine Adipic acid Homogentisic acid 2-ketoglutaric acid Malic acid Citric acid Malonic acid Ethylmalonic acid N-(3-Phenylpropionyl) glycine Glutaconic acid N-Acetylaspartic acid Glutaric acid N-Acetyltyrosine Lactic acid N-Isovalerylglycine Methylmalonic acid Oxoproline Orotic acid 4-OH-Phenyllactic acid Phenylpyruvic acid Propionylglycine Pyruvic acid Sebacic acid Succinic acid -
Extracted LC-MS/MS Chromatogram of Panel 1 and Panel 2
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Results
Number
Organic Acids
LOQ (ppm)
(%) RSD
1
Glycolic acid
0.72
6.64
2
Malic acid
0.06
4.54
3
Malonic acid
0.07
6.91
4
Fumaric acid
0.06
9.67
5
N-Acetylaspartic acid
0.10
4.67
6
Oxoproline
0.05
3.53
7
2-Oxoadipic acid
0.06
6.50
8
2-Methylcitrate
0.72
29.64
9
Propionylglycine
0.18
3.36
10
3-OH-3-Methylglutaric acid
0.18
1.74
11
2-OH Butyric acid
0.06
5.86
12
3-OH Butyric acid
0.26
9.17
13
3-OH Isobutyrate
0.55
0.71
14
Homogentisic acid
0.04
2.26
15
4-OH-Phenyllactic acid
0.08
10.32
16
N-Acetyltyrosine
0.13
4.53
17
3-Methylglutaconic acid
0.05
2.92
18
Succinylacetone
0.05
5.19
19
N-Isovalerylglycine
0.07
3.26
20
Suberylglycine
0.13
2.26
21
2-OH-Phenylacetic acid
0.07
1.80
22
4-OH Phenyl acetic acid
1.40
1.58
23
3-Phenyllactic acid
0.06
6.14
24
N-(3-Phenylpropionyl) glycine
0.05
1.97
25
Hexanoylglycine
0.09
3.07
26
Suberic acid
0.08
1.44
27
3-OH-2-Methylbutanoic acid
0.09
12.17
28
Sebacic acid
0.07
2.93
29
Pyruvic acid
0.22
12.45
30
2-Ketoglutaric acid
0.61
1.35
31
Citric acid
0.39
3.68
32
Orotic acid
0.04
2.98
33
Lactic acid
3.54
10.83
34
2-OH-Glutaric acid
0.37
0.73
35
3-OH-Propanoic acid
1.49
3.66
36
Methylmalonic acid
0.34
2.84
37
3-OH-Glutaric acid
0.05
1.17
38
Succinic acid
0.10
1.92
39
Ethylmalonic acid
0.05
2.07
40
Glutaconic acid
0.20
6.07
41
4-OH-Phenylpyruvic acid
0.39
4.88
42
3-OH-Isovaleric acid
0.11
2.64
43
Glutaric acid
0.23
1.73
44
3-Methyl-2-Oxovaleric acid
0.13
6.15
45
2-OH-Isovaleric acid
0.07
2.46
46
3-OH-Pentanoic acid
0.16
4.06
47
4-Methyl-2-oxovaleric acid
0.15
4.44
48
Tiglylglycine
0.05
3.17
49
3-Methylcrotonyl glycine
0.10
2.47
50
3-Methylglutaric acid
0.08
4.07
51
Phenylpyruvic acid
0.10
3.12
52
Adipic acid
0.08
4.00
53
2-OH-3-Methylpentanoic
0.08
1.46
