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ORGANIC ACIDS

ORGANIC ACID IN URINE
LC-MS/MS KIT

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. “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.

  • Highlights

    • 12 min. run for each panel (Total analysis time 24 minutes-Panel 1&2)

    • Analysis of 54 organic acids without derivatization

    • Quick and easy sample preparation; just dilution

    • Successful chromatographic separation of isomers

    • Consuming small volume of patient’s sample

  • Matrıx

    • Urine

  • Parameters

    Please see the panels below

Sample Preparation for Urine

  • Step 1

    Dilute urine sample 5 times using Reagent-1

  • Step 2

    Transfer 50 µl of diluted urine sample to a vial

  • Step 3

    Add 25 µl of IS mix and 425 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

    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

  • 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
ADAPTING DIAGNOSTICS IN CHROMATOGRAPHY COUPLED MASS SPECTROMETRY..

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