AMINO ACIDS

AMINO ACIDS
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. Diseaserelated 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 alloIsoleucine 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

    Quantitative Amino Acids Analysis

    Alanine, Arginine, Aspartic acid, Asparagine, 2-Aminobutyric acid, Argininosuccinic acid, Anserine,
    2-Aminoadipic acid, 3-Aminoisobutyric acid, β-Alanine, Ethanolamine, Phenylalanine, orthoPhosphoserine, ortho-Phosphorylethanolamine, β-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

    • Serum
    • 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 HPLCgrade 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

Explore Results

NO

AMINO ACIDS

LOQ (nmol/ml)

Recovery (%)

Repeatability (%)

1

Tryptophan

0.23

117

3.68

2

Taurine

5.47

85

4.81

3

Phenylalanine

0.73

110

3.73

4

Tyrosine

3.34

101

4.09

5

Leucine

0.20

97

4.62

6

Isoleucine

0.22

107

1.13

7

Methionine

0.41

90

3.70

8

Gamma-aminobutyric acid

0.07

104

8.49

9

3-Amino-isobutyric acid

0.15

100

6.49

10

Norvaline

0.15

95

4.90

11

2-Amino-adipic acid

0.83

94

2.23

12

Beta alanine

1.00

90

7.22

13

Ethanolamine

0.56

106

5.79

14

Aspartic acid

1.06

96

3.96

15

L-2-aminobutyric acid

0.22

94

4.71

16

Threonine

0.72

97

3.99

17

Serine

0.89

100

5.39

18

Alanine

6.82

104

4.44

19

Glycine

9.08

101

2.24

20

Asparagine

0.46

91

3.71

21

Trans-4-hydroxy-L-proline

0.28

102

1.90

22

Glutamine

0.32

98

3.58

23

Sarcosine

4.38

113

2.16

24

Proline

0.67

100

3.66

25

Homocitrulline

0.89

105

3.50

26

Citrulline

0.51

93

3.42

27

Homocysteine

0.31

103

5.32

28

Cystine

0.97

100

4.39

29

Cystathionine

0.25

104

5.81

30

Arginine

0.15

107

3.66

31

Histidine

2.43

54

1.49

32

Ornithine

1.39

96

3.34

33

DL-5- hydroxylysine

0.30

104

1.64

34

Lysine

0.09

114

4.52

35

Carnosine

0.12

93

2.56

36

3-Methylhistidine

0.30

100

4.49

37

1-Methylhistidine

0.20

78

2.94

38

Anserine

0.21

107

3.72

39

O-phosphoserine

0.12

99

6.20

40

Glutamic acid

0.24

98

9.30

41

Valine

5.11

42

4.68

42

Argininosuccinic acid

0.62

87

3.75

43

Orthophosphoryl ethanolamine

2.56

90

2.96

44

Allo-isoleucine

0.04

101

1.51


  • Extracted chromatogram of amIno acIds-spIked serum sample

ADAPTING DIAGNOSTICS IN CHROMATOGRAPHY COUPLED MASS SPECTROMETRY..

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