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47 Amino Acids LC-MS/MS Analysis Kit

47 Amino Acids LC-MS/MS Analysis Kit
CE-IVD certified and suitable for any kind of LC-MS/MS instrument

 

The Zivak 47 Amino Acids LC-MS/MS analysis kit can be used in combination with the Zivak-Multitasker as well as manually. Analysis run time for the manual version is 20 minutes. Due to the efficient and quite smart procedure of the Zivak-Multitasker, the automated version guarantees to receive quantitative results in just 20 minutes per test, including sample preparation, sample injection and analysis run time. Additionally, high accuracy is being rectified through external quality tests.

 

Quantitative LC-MS/MS analysis kit for Amino Acids in biological fluids (Plasma & Serum & Urine & Cerebrospinal Fluid). 

Amino acids are the basic structural units of proteins. Quantitative analysis of amino acids in biological fluids is one of the most important parts of the clinical diagnosis of patients with inborn errors of amino acid metabolism or monitoring healthy people. Amino acid analysis in urine is important for diagnosis of disorders which are related with renal amino acid transport. Quantitative analysis of amino acids in plasma is preferred over urine for the diagnosis of aminoacidopathies which cause to accumulation or the deficiency of amino acids in biological fluids.

Quantitative analysis of amino acids in biological fluids also has clinical importance for determining some neurological and endocrinal disorders. LC-MS/MS is an influential device for the study of disorders related with amino acids. LC-MS/MS analysis gives results for 47 free amino acids in 20 minutes.

Zivak Quantitative Free Amino Acids LC-MS/MS Analysis Kit was developed for quantitative detection of free amino acids in human serum, plasma, urine and cerebrospinal fluid LC-MS/MS is an influential device for the study of metabolic disorders and LC-MS/MS analysis gives results in 20 minutes with minimal sample preparation. Main methods and procedures that have been selected are based on EN ISO 13485 and 98/79/EC.

 

47 Amino Acids LC-MS/MS Analysis Kit 0 47 Amino Acids LC-MS/MS Analysis Kit 1 47 Amino Acids LC-MS/MS Analysis Kit 2 47 Amino Acids LC-MS/MS Analysis Kit 3 47 Amino Acids LC-MS/MS Analysis Kit 4 47 Amino Acids LC-MS/MS Analysis Kit 5 47 Amino Acids LC-MS/MS Analysis Kit 6 47 Amino Acids LC-MS/MS Analysis Kit 7 47 Amino Acids LC-MS/MS Analysis Kit 8
LC-MS/MS Parameters

Device

LC-MS/MS Analysis System

Column

Zivak Amino Acid Biological Fluids LC-MS/MS Analytical Column

Injection Volume

3 µL

Pump Program

00:00  min                   35 % B

14:00  min                   55 % B

15:00  min                   95 % B

16:30  min                   95 % B

16:31  min                   35 % B

20:00  min                   35 % B

Flow

0.25 mL/min

Ionization Mode

ESI positive+negative

CID Gas

2.4 mTorr

API Nebulizing Gas

55 psi

Scan Time

2,62 sec

SIM Width

1.5 amu

Drying Gas

35 psi

Drying Gas Temp.

350 °C

Needle

5000V

Shield

600V

Capillary

50V

Detector

1500 V

Column Oven

25C

 
Analytical Performance

No

Analyte

LOD (µmol/L)

LOQ (µmol/L)

Accuracy (%)

Inter-Assay Precision  (%CV)

Linearity

(R2)

1st Day

2rd  Day

3th Day

1

3-methyl-histidine (3-MeHIS)

0,016

0,051

91,98

3,12

3,57

5,90

0,96059

2

Alanine (ALA)

0,004

0,012

105,98

2,03

3,70

3,22

0,99498

3

Alpha-aminoadipic acid (AAA)

0,067

0,222

90,52

2,93

6,20

2,89

0,96194

4

Alpha-aminobutyric acid (ABA)

0,031

0,102

96,75

5,91

5,09

5,22

0,98165

5

Alpha-aminopimelic acid (APA)

0,001

0,002

104,66

6,27

7,08

3,26

0,97653

6

Anserine ( ANS)

0,025

0,081

101,09

12,16

14,43

12,06

0,91248

7

Arginine (ARG)

0,007

0,023

92,45

3,56

5,89

6,12

0,99633

8

Asparagine  (ASN)

0,027

0,089

94,77

6,66

5,04

6,74

0,99824

9

Aspartic Acid (ASP)

0,006

0,019

85,64

4,82

3,83

3,41

0,99827

10

Beta alanine (BALA)

0,003

0,009

93,64

6,81

8,25

5,89

0,92526

11

Beta-aminoisobutyric acid (BAIB)

0,001

0,002

89,64

7,18

7,47

5,66

0,99123

12

Citrulline (CIT)

0,028

0,092

101,15

3,10

3,42

2,75

0,99917

13

Cystathionine (CTH)

0,001

0,005

87,81

3,99

2,19

1,64

0,99807

14

Gamma-aminobutyric acid (GABA)

0,182

0,601

94,81

11,76

7,47

7,01

0,88421

15

Glutamic Acid (GLU)

0,001

0,002

83,42

5,45

6,92

3,99

0,99867

16

Glutamine (GLN)

0,001

0,002

106,09

7,51

2,59

6,04

0,99849

17

Glycine (GLY)

0,000

0,001

93,53

6,15

5,68

5,87

0,99836

18

Histamine (HISTA)

0,002

0,007

101,35

2,11

1,78

4,79

0,99882

19

Histidine (HIS)

0,008

0,026

101,63

3,99

3,58

4,82

0,99940

20

Hydroxylysine (HYL)

0,013

0,043

91,95

4,59

2,83

6,04

0,98041

21

Hydroxyproline (HYP)

0,002

0,005

96,86

5,31

4,01

5,96

0,99719

22

Isoleucine (ILEU)

0,092

0,302

90,28

0,59

2,82

0,73

0,99886

23

Leucine (LEU)

0,025

0,084

90,52

1,97

5,01

3,08

0,99561

24

Lysine (LYS)

0,014

0,047

101,10

0,62

1,79

2,43

0,99938

25

Methionine (MET)

0,023

0,075

103,09

3,56

3,66

3,97

0,99943

26

N-methyl-histidine (N-MeHIS)

0,008

0,028

93,09

7,11

5,54

7,57

0,98831

27

Ornithine hydrochloride (ORN)

0,009

0,030

94,27

3,44

4,21

3,06

0,99934

28

Phenylalanine (PHE)

0,029

0,094

99,73

5,49

4,15

4,85

0,98135

29

Proline (PRO)

0,004

0,012

98,56

1,93

1,82

2,14

0,95264

30

Sarcosine (SAR)

0,008

0,025

94,17

5,81

6,68

4,53

0,99634

31

Serine (SER)

0,006

0,020

90,94

5,56

5,86

4,09

0,97524

32

Serotonin (SRTN)

0,009

0,029

100,46

3,66

1,70

3,81

0,98610

33

Thiaproline (THPR)

0,008

0,027

88,19

5,33

5,69

6,55

0,98451

34

Threonine (THR)

0,003

0,009

91,72

2,06

3,24

2,69

0,98219

35

Trytophan (TRP)

0,004

0,013

104,62

5,15

4,05

4,97

0,95469

36

Tyrosine (TYR)

0,003

0,009

100,10

2,52

3,41

2,02

0,99838

37

Valine (VAL)

0,033

0,108

99,00

3,52

4,36

2,98

0,99928

38

Taurin (TAU)

0,018

0,058

104,11

8,20

6,63

7,42

0,99238

39

Carnosine (Car)

0,015

0,050

85,01

9,12

9,60

7,17

0,95443

40

5-OH-Trp

0,037

0,121

98,95

12,47

12,24

10,89

0,96086

41

O-Phospho-L-Serine (PS)

0,004

0,012

86,44

8,62

7,81

12,89

0,97123

42

Creatinine (CRN)

0,017

0,056

99,22

3,16

5,18

4,26

0,98485

43

N-acetyl-L-tyrosine (NAT)

0,350

1,154

98,68

4,11

5,05

6,09

0,91113

44

Cysteine (CYS)

0,010

0,032

90,29

3,15

3,01

2,11

0,97364

45

Homocysteine (HCYS)

0,014

0,047

95,11

2,54

3,44

4,18

0,98114

46

Cystine (C-C)

0,001

0,002

94,38

3,77

4,08

3,36

0,98226

47

Homocystine (HC-HC)

0,0001

0,0002

100,66

4,07

2,16

3,27

0,97630

 
 

Storage & Stability

- This analysis kit can be shipped at room temperature.

- After arrival, standards and controls should be stored at 2-8 °C.

- Reagent 1, 2, 3, 4 should be stored at 2-8 °C.

- All other components of the kit should be stored at between room temperature.

- Reagent 2, 3 and 4 are volatile, please make sure that the lids of reagent bottles are closed tightly after use.

- All components are guaranteed until expiry date when stored at recommended temperatures and used as described in these instructions.

Preparation of Calibrators & Controls:

Add 3 ml of deionized water to lyophilised plasma calibrators and controls and dissolve.

 After reconstitution, plasma calibrator must be aliquoted into 150 µl volumes and stored at -20°C.

Note: Please use zivak lyophilized sulfur group calibrators, control for Homocysteine, cysteine, cystine, homocystine. Zivak lyophilized sulfur group calibrators contain homocysteine, cysteine, cystine, homocystine.

 

Sample Pretreatment (Manual):

Take 100 mL of patient sample (or calibrators and controls) into the sample preparation tube.            

Add 100 mL of Reagent 1 and vortex for 10 seconds.

Add 200 mL of Reagent 2 and vortex for 10 seconds.

Add 200 mL of Reagent 3 and vortex for 10 seconds. Wait for 2 minutes and vortex again for 10 seconds. Wait for 1 minute.

Add 200 mL of Reagent 4 and vortex for 10 seconds. After mixing wait for a minute.

Centrifuge at 5.000 x rpm for 3 minutes.

Take 100 mL the upper phase into the autosampler vial.

Inject 3 mL to the LC-MS/MS system.

Note: The prepared sample is stable at 2-8 °C for 24 hours.

 

Sample Pretreatment (Manual) for Homocysteine, Cysteine:

Take 100 mL of patient sample (or calibrators and controls) into the sample preparation tube.

Add 100 mL of Reagent H and vortex for 10 seconds. Wait for 1 minute and vortex again for 10 seconds.

Add 100 mL of Reagent 1 and vortex for 10 seconds.

Add 200 mL of Reagent 2 and vortex for 10 seconds.

Add 200 mL of Reagent 3 and vortex for 10 seconds. Wait for 2 minutes and vortex again for 10 seconds. Wait for 1 minute.

Add 200 mL of Reagent 4 and vortex for 10 seconds. After mixing wait for a minute.

Centrifuge at 5.000 x rpm for 3 minutes.

Take 100 mL the upper phase into the autosampler vial.

Inject 3 mL to the LC-MS/MS system.

Note: The prepared sample is stable at 2-8 °C for 24 hours.

 

Sample Pretreatment (Automated):

Take 100 mL of patient sample (or calibrators and controls) into the Zivak Multitasker Sample Preparation vials

Make sure that reagents and wash solution are connected to the reagent pump and purged properly. 

Place the vials on the MULTITASKER system tray.

Note: The prepared sample is stable at 2-8°C for 24 hours.

 

Sample Pretreatment (Automated) for Homocysteine, Cysteine:

Take 100 mL of patient sample (or calibrators and controls) into the Zivak Multitasker Sample Preparation vials

Make sure that reagents and wash solution are connected to the reagent pump and purged properly. 

  Place the vials on the MULTITASKER system tray. (Please use Aminoacids HCYS automated method)

Note: The prepared sample is stable at 2-8°C for 24 hours.

Materials Supplied

Order No.

Volume

Component

ZV-3002-02R1-10

1 x 30 ml

Reagent 1, Contains deuterated internal standards

ZV-3002-02R2-10

1 x 50 ml

Reagent 2, Contains base

ZV-3002-02R3-10

1x 50 ml

Reagent 3, Contains derivatization reagent

ZV-3002-02R4-10

1 x 50 ml

Reagent 4, Contains organic modifier

ZV-3002-02RH-10

1 x 30 ml

Reagent H, Protective Reagent for HCYS

ZV-3002-02MA-10

1 x 1 L

Mobile Phase A, Contains organic solvent

ZV-3002-02MB-10

1 x 1 L

Mobile Phase B, Contains organic solvent

ZV-3002-02WB-10

1 x 1 L

Washing solution, Contains organic solvent

ZV-3002-KK-10

1x1 pc

User Guide

 
 
Materials Required But Not Supplied

Order No.

Volume

Component

ZV-3002-02S1-10

1 x 3 ml

Lyophilized Plasma Calibrator Level-1

ZV-3002-02S2-10

1 x 3 ml

Lyophilized Plasma Calibrator Level-2

ZV-3002-02S3-10

1 x 3 ml

Lyophilized Plasma Calibrator Level-3

ZV-3002-02K1-10

1 x 3 ml

Lyophilized Plasma Control Level 1

ZV-3002-02K2-10

1 x 3 ml

Lyophilized Plasma Control Level 2

ZV-3002-02K3-10

1 x 3 ml

Lyophilized Plasma Control Level 3

ZV-3002-02SGS1-10

1 x 3 ml

Sulfur Group Lyophilized Plasma Calibrator Level 1

  ZV-3002-02SGS2-10

1 x 3 ml

Sulfur Group Lyophilized Plasma Calibrator Level 2

  ZV-3002-02SGK1-10

1 x 3 ml

Sulfur Group Lyophilized Plasma Control Level 1

  ZS-2ML-8-1001

2x100pcs

2 ml glass autosampler vial for automated Sample Preparation

  ZS-9004-0100-00

2x100pcs

2 ml glass autosampler vial caps pre-slit for automated Sample Preparation

ZV-3002-02C1-10

1 x 1 pcs

Zivak Amino Acid Biological Fluids LC-MS/MS Analytical Column

 
File Name Type Size Download
User Manual 1.19 MB

Zivak General Catalog

Download Zivak General Catalog PDF