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AMINO ACIDS

Amino Acids in Plasma/Urine LC-MS/MS Analysis Kit

In recent years metabolomics has facilitated gaining insight into discovering the potential disease diagnostic markers and observing fluctuations of certain metabolite levels in body fluids. Disease-related metabolites can be uncovered by following progress of the new methodologies which enable to identify a broad range of diagnostic biomarker candidates including amino acids. Owing to the vital roles of amino acids in metabolic regulations (protein synthesis, neurotransmission, hormone secretion, and immunity), they have become significant targets while taking snapshot of metabolic status. Therefore, elucidation of the circulating amino acids profile alterations by utilizing robust analysis method could shed light on not only in metabolic diseases but also in certain disorders such as autism, prostate cancer, Huntington’s disease, Parkinson’s disease, Alzheimer’s disease, obesity, type 2 diabetes, liver cirrhosis, major depressive disorder, and cardiovascular disease.

  • Highlights

    Quantitative analysis of underivatized 43 amino acids in 7.5 min. In need of determining allo-Isoleucine for MSUD, extra injection takes 11.0 min. (Leucine/Isoleucine/allo-Isoleucine).

    One sample preparation for all amino acids-one vial for both abovementioned methods.

    Just a few pipetting steps for the sample preparation; for plasma/serum-protein crash and shoot for urine-dilute and shoot.

    Clinically important amino acids safeguarded by its own internal standard.

    Consuming small volume of patient’s sample.

    Long life span of HPLC column-up to 10000 injections.

  • Parameters

    Alanine, Arginine, Aspartic acid, Asparagine, 2-Aminobutyric acid, Argininosuccinic acid, Anserine, 2-Aminoadipic acid, 3-Aminoisobutyric acid, β-Alanine, Ethanolamine, Phenylalanine, ortho-Phosphoserine, ortho-Phosphorylethanolamine, γ-Aminobutyric acid, Glutamic acid, Glycine, Glutamine, trans-4-Hydroxyproline, Homocitrulline, Homocystine, Histidine, 5-Hydroxylysine, Isoleucine, Carnosine, Lysine, Leucine, Allo-Isoleucine, Methionine, 3-Methylhistidine, 1- Methylhistidine, Norvaline, Ornithine, Proline, Cystine, Cystathionine, Sarcosine, Citrulline, Serine, Taurine, Tryptophan, Tyrosine, Threonine and Valine.

  • Matrix

    Plasma/Urine

Sample Preparation

Plasma/Serum
  • Step 1
    Pipette 50 µL of serum/plasma patient’s sample (calibrator/control) into a glass centrifuge tube.
  • Step 2
    Add 50 µL of internal standard solution and vortex for 5 sec.
  • Step 3
    Add 700 µL of Reagent-1, vortex for 5 sec. then, centrifuge at 4000 rpm for 5 min.
  • Step 4
    Decant the clear supernatant into HPLC vial prior to injection.

Sample Preparation

Urine
  • Step 1
    Pipette 100 µL of urine sample into a glass centrifuge tube and dilute with 900 µL of HPLC-grade water.
  • Step 2
    Pipette 50 µL of diluted urine sample into a glass centrifuge tube.
  • Step 3
    Add 50 µL of internal standard solution and vortex for 5 sec.
  • Step 4
    Add 700 µL of Reagent-1, vortex for 5 sec. then, centrifuge at 4000 rpm for 5 min.
  • Step 5
    Decant the clear supernatant into HPLC vial prior to injection.

  • Chromatogram

  • METHOD PERFORMANCE

    Analyte

    LOQ (µmol/L) 6460 QQQ

    Accuracy % 6460 QQQ

    Repeatabilty % CV 6460  QQQ

    Linearity R2 646 QQQ

    1-Methylhistidine

    0,2

    85,3

    2,94

    0,9996

    2-Aminoadipic acid

    0,83

    93,67

    2,37

    0,9992

    2-Aminobutyric acid

    0,22

    93,67

    4,12

    0,9998

    3-Aminoisobutyric acid

    0,15

    98,93

    7,55

    0,9995

    3-Methylhistidine

    0,3

    99,87

    4,19

    0,9992

    4-Aminobutyric acid

    0,07

    102,62

    5,41

    0,9994

    5-Hydroxylysine

    0,3

    102,78

    1,54

    0,9988

    Alanine

    6,82

    105,04

    4,19

    0,9988

    Allo-isoleucine

    0,22

    98,3

    1,65

    0,9998

    Anserine

    0,21

    106,86

    4,13

    0,9998

    Arginine

    0,15

    103,88

    3,98

    0,9998

    Argininosuccinic acid

    0,62

    93,04

    2,97

    0,9981

    Asparagine

    0,46

    86,17

    3,09

    0,9997

    Aspartic acid

    1,06

    91,19

    3,44

    0,9956

    Beta-Alanine

    1,0

    91,84

    9,11

    0,999

    Carnosine

    0,12

    96,08

    3,97

    0,9984

    Citrulline

    0,51

    82,47

    3,44

    0,9977

    Cystathionine

    0,25

    104,65

    4,75

    0,9997

    Cystine

    0,97

    95,82

    4,63

    0,9998

    Ethanolamine

    0,56

    104,25

    5,67

    0,9944

    Glutamic acid

    0,24

    97,88

    6,1

    0,999

    Glutamine

    0,32

    98,03

    3,37

    0,9999

    Glycine

    9,03

    95,82

    1,93

    0,9984

    Histidine

    2,43

    107

    1,12

    0,9999

    Homocitrulline

    0,89

    104,4

    3,71

    0,9993

    Homocystine

    0,31

    102,82

    4,8

    0,999

    Isoleucine

    0,22

    110,92

    1,39

    0,9991

    Leucine

    0,2

    92,22

    4,85

    0,9997

    Lysine

    0,09

    113,46

    5,12

    0,9988

    Methionine

    0,41

    87,42

    1,88

    0,9999

    Norvaline

    0,15

    93,51

    4,94

    0,9988

    Ornithine

    1,39

    93,34

    3,54

    0,9996

    O-Phosphorylethanolamine

    2,56

    87

    4,28

    0,9976

    O-Phosphoserine

    0,12

    83,96

    4,49

    0,9969

    Phenylalanine

    0,73

    108,04

    3,05

    0,997

    Proline

    0,67

    99,84

    3,3

    0,9998

    Sarcosine

    4,38

    110,03

    2,25

    0,9964

    Serine

    0,89

    99,26

    4,58

    0,997

    Taurine

    5,47

    85,09

    5,28

    0,9956

    Threonine

    0,72

    95,03

    3,27

    0,9999

    trans-4-hydroxyproline

    0,28

    87,8

    1,9

    0,9914

    Tryptophan

    0,23

    109,81

    3,21

    0,9998

    Tyrosine

    3,34

    101,19

    3,45

    0,9991

    Valine

    5,11

    98,65

    6,08

    0,9986
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