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25-Hydroxyvitamin D2 / D3 Serum LC-MS/MS APCI Analysis Kit

 25-Hydroxyvitamin D2 / D3 Serum LC-MS/MS APCI Analysis Kit
CE-IVD certified and suitable for any kind of LC-MS/MS instrument

The Zivak Vitamin D2/D3 analysis kit can be used in combination with the Zivak-Multitasker as well as manually. Analysis run time for the manual version is 1.7 minutes. Due to the efficient and quite smart procedure of the Zivak-Multitasker, the automated version guarantees also to receive results in 1.7 minutes per test, including sample preparation-, sample injection- and analysis run time. Additionally, high accuracy is being rectified through NIST (National Institute of Standards and Technology) with an accuracy of 96%.

 
 
Quantitative LC-MS/MS analysis kit for 25-hydroxy metabolites of Vitamin D2 and D3 in human serum samples.

Vitamin D, commonly known as the “sunshine vitamin”, is essential for maintaining calcium metabolism and bone health. The major physiologic function of Vitamin D is to maintain the bloodcalcium and phosphorus levels within the normal range in order to maintain metabolic functions that are essential for most life processes, Vitamin D3 and Vitamin D2 are metabolised to 25-OH-Vitamin D3 and 25-OH-Vitamin D2 in the liver by the 25-OHase enzyme. The quantitation of these metabolites is widely used for determining of Vitamin D status in human. The Vitamin D status has a clinical significance in a variety of disorders which related to hormones. The measurements of the serum concentration of 25-OH-Vitamin D3 and 25-OH-Vitamin D2 have been used for differential diagnosis of hypocalcemic and hypercalcemic metabolic bone disorders.

The analysis of these metabolites is considered to be difficult. Some reasons of the difficulties are below;

  • The low concentrations of 25-OH Vitamin D metabolites in serum. (level of µg/L)

  • The low stability in the presence of light and heat.

  • The presence of other metabolites which have chemical similarity.

  • The large amount of other related neutral lipids (such as cholesterol)

25-OH Vitamin D2-D3 LC-MS/MS Analysis Kit was developed for rapid, sensitive and reliable quantitative detection of the 25-OH Vitamin D2-D3 concentrations in human serum samples and it gives results in 1.7 minutes with minimal sample preparation. Main methods and procedures that we selected are based on EN ISO 13485 and 98/79/EC.

 25-Hydroxyvitamin D2 / D3 Serum LC-MS/MS APCI Analysis Kit 0
LC-MS/MS Parameters
Instrument Zivak Tandemgold LC-MS/MS System
Column Zivak 25-Hydroxyvitamin D2-D3 Serum LC-MS/MS APCI Analytical Column
Injection Volume 50 μl
Ionization Mode APCI Positive
CID Gas 2.4 mTorr
API Nebulizing Gas 55 psi
Vaporizer Gas 15 psi
Vaporizer Gas Temperature 350 ºC
Drying Gas 25 psi
Drying Gas Temperature 300 ºC
Spray Chamber Temperature 60 ºC
SIM Width 1.5 amu
Scan Time 0.600 seconds
Corona Current 10 μA
Shield Voltage 600 V
Detector Voltage 1600 V

 

No

Analyte

Accuracy

(%)

Intra-assay Precision

(%CV)

Inter-assay Precision

 (%CV)

1

25-OH Vitamin D3

94.7

2.16

2.74

2

25-OH Vitamin D2

96.4

4.60

4.74

 

No

Analyte

LOD

(µg/L)

LOQ

(µg/L)

Linearity

(R2)

1

25-OH Vitamin D3

1.48

4.90

0.993

2

25-OH Vitamin D2

1.06

3.50

0.992

•   This analysis kit can be shipped at room temperature.

•   Serum controls, calibrators and reagent 2 must be stored at 2-8 °C until reconstitution.

•   After arrival, mobile phases and reagent 1 must be stored at room temperature.

•   After reconstitution serum calibrators and controls must be stored at -20 °C.

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

 

Preparation of Calibrators:

  • Add 3 mL of deionized water to each serum calibrator and dissolve.
  • After preparation, they must be aliquoted into 450 µL volumes and stored at -20°C.

Preparation of Controls:

  • Add 3 mL of deionized water to each serum control and dissolve.
  • After preparation, they must be aliquoted into 450 µL volumes and stored at -20°C.

 

Sample Pretreatment (Manual)

  1. 1. Take 400 µL of patient sample (or calibrators and controls) into the sample preparation tube.
  2. 2. Add 450 µL Reagent 1 and vortex for 10 seconds.
  3. 3. Add 400 µL Reagent 2 and vortex for 10 seconds.
  4. Centrifuge at 5000 rpm x 3 minutes.
  5. 0.5 mL Take the upper phase to vial.
  6. Inject 50 µL to LC-MS/MS system.

Prepared samples are stable for 24 hours at 2-4 °C temperature.

 

Sample Pretreatment (Automated)

  1. If Calibrator/Control will be analyzed, take 400 µL of calibrator/control sample into glass vial and put vials into related tray on instrument.
  2. Place serum samples into system tray on Zivak Multitasker Full Automated Sample Preparation and Injection System.
  3. Analysis will automatically start after clicking Start button on software.
  4. Injections, calculations and reporting carried out automatically by the instrument.

Prepared samples are stable for 24 hours at 2-4 °C temperature.

Materials Supplied

Order No

Volume

Component

ZV-3046-02R1-10

3 x 45 mL

Reagent 1,

Contains aqueous buffer

ZV-3046-02R2-10

3 x 45 mL

Reagent  2,

Contains Internal Standard

ZV-3046-02MP-10

1 x 0.5 L

Mobile Phase A,

Contains organic solvent

ZV-3046-02MT-10

1 x 0.5 L

SPE Buffer,

Contains organic solvent

ZV-3046-02WB-10

2 x 1 L

Wash Solution

ZV-4027-KK-10

1 x 1 pc

User Guide

 
 
Materials Required But Not Supplied

Order No

Volume

Component

ZV-3046-02S1-10

1 x 3 mL

Lyophilized, Serum Calibrator Level-1

ZV-3046-02S2-10

1 x 3 mL

Lyophilized, Serum Calibrator Level-2

ZV-3046-02S3-10

1 x 3 mL

Lyophilized, Serum Calibrator Level-3

ZV-3046-02S4-10

1 x 3 mL

Lyophilized, Serum Calibrator Level-4

ZV-3046-02S5-10

1 x 3 mL

Lyophilized, Serum Calibrator Level-5

ZV-3046-02S6-10

1 x 3 mL

Lyophilized, Serum Calibrator Level-6

ZV-3046-02K1-10

1 x 3 mL

Lyophilized, Serum Control Level-1

ZV-3046-02K2-10

1 x 3 mL

Lyophilized, Serum Control Level-2

ZV-3046-02C1-10

1 x 1 pc

25-Hydroxyvitamin D2-D3 Serum LC-MS/MS  APCI Analytical Column

ZV-3046-CSPE-10

1 x 1 pc

25-Hydroxyvitamin D2-D3 Serum LC-MS/MS Trap Column Cartridge

ZV-3046-02C2-10

1 x 1 pc

25-Hydroxyvitamin D2-D3 Serum LC-MS/MS Trap Column Holder

ZS-9020-0001-00

 

200 – 1000 µL pipette

ZS-9007-0500-00

 

200 – 1000 µL pipette tips

ZS-2ML-8-1001

2 x 100 pc

2 mL glass vial

ZS-9004-0100-00

2 x 100 pc

2 mL glass vial caps pre-slit

 

Product Video

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User Manual 1010 KB

Zivak General Catalog

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