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25-Hydroxyvitamin D2-D3 Serum VD-200 UHPLC Analysis Kit

25-Hydroxyvitamin D2-D3 Serum VD-200 UHPLC Analysis Kit

25-OH Vitamin D2-D3 VD-200 UHPLC 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 6 minutes with minimal sample preparation. Main methods and procedures that have been selected are based on EN ISO 13485 and 98/79/EC.

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 blood calcium 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 metabolized 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-Hydroxyvitamin D2-D3 Serum VD-200 UHPLC Analysis Kit 0
HPLC-UV Parameters

Instrument

Zivak VD-200 Full Automated Vitamin D2/D3 UHPLC Analyzer

Column

Zivak 25-Hidroxyvitamin D2-D3 Serum VD-200 UHPLC Analytical Column

SPE

Zivak 25-Hidroxyvitamin D2-D3 Serum VD-200 UHPLC Trap Column Cartridge

Detector

UV/VIS

Wavelength

264 nm

Injection Volume

20 µL

Column Oven Temperature

35 °C

 
 
 
Analytical Performance

No

Analyte

Accuracy

(%)

Intra-assay Precision

(%CV)

Inter-assay Precision

 (%CV)

 

 

 

1

 

 

 

25-OH Vitamin D3

 

Low level

 

High level

Low level

High level

Low level

High level

97.171

100.878

3.476

1.998

3.248

2.744

2

25-OH Vitamin D2

94.196

100.811

3.381

1.815

3.684

2.470

 

No

Analyte

LOD

(µg/L)

LOQ

(µg/L)

Linearity

(R2)

1

25-OH Vitamin D3

1.65

5.00

1. Day

2. Day

3. Day

0,9989

0,9979

0,9986

2

25-OH Vitamin D2

1.80

5.45

0,9984

0,9951

0,9974

Storage & Stability:

  • This analysis kit can be shipped at room temperature.
  • Calibrator and controls (after reconstitution) must be stored at -20 °C.
  • All the other components of the kit should be stored at room temperature.
  • Please make sure that you keep reagent 1 at room temperature otherwise it will become unusable.
  • 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 each serum calibrator and dissolve.
  • After preparation, they must be aliquoted into 400 µL volumes and stored at        -20°C.

 

Sample Pretreatment (Automated):

If Calibrator/Control will be analyzed, take 400 µL of calibrator/control sample into glass vial and put vials into related tray on instrument.

 

Place serum samples into system tray.

 

Analysis will automatically start after clicking Start button on software.

                                                                           

Injections, calculations and reporting done automatically by instrument.

 

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

Materials Supplied

Order No

Volume

Component

ZV-4027-05R1-10

1 x 350 mL

Reagent 1, Contains aqueous buffer

ZV-4027-05R2-10

1 x 300 mL

Reagent  2, Contains Internal Standard

ZV-4027-05MP-10

2 x 1.5 L

Mobile Phase A, Contains organic solvent

ZV-4027-05MT-10

2 x 1.5 L

SPE Buffer, Contains organic solvent

ZV-4027-05W1-10

1 x 2 L

Wash Solution 1

ZV-4027-05W2-10

2 x 2 L

Wash Solution 2

ZS-2ML-8-1001

5 x 100 pcs

2 mL glass vial

ZS-9004-0100-00

5 x 100 pcs

2 mL glass vial caps pre-slit

ZV-4027-KK-10

1 x 1 pc

User Guide

 
 
Materials Required But Not Supplied

Order No

Volume

Component

ZV-4027-02S1-10

1 x 3 mL

Lyophilized, Serum Calibrator Level-1

ZV-4027-02S2-10

1 x 3 mL

Lyophilized, Serum Calibrator Level-2

ZV-4027-02S3-10

1 x 3 mL

Lyophilized, Serum Calibrator Level-3

ZV-4027-02S4-10

1 x 3 mL

Lyophilized, Serum Calibrator Level-4

ZV-4027-02S5-10

1 x 3 mL

Lyophilized, Serum Calibrator Level-5

ZV-4027-02S6-10

1 x 3 mL

Lyophilized, Serum Calibrator Level-6

ZV-4027-02K1-10

1 x 3 mL

Lyophilized, Serum Control Level-1

ZV-4027-02K2-10

1 x 3 mL

Lyophilized, Serum Control Level-2

ZV-4027-02C1-10

1 x 1 pc

25-Hydroxyvitamin D2-D3 Serum VD-200 UHPLC Column

ZS-9020-0001-00

 

200 – 1000 µL pipette

ZS-9007-0500-00

 

200 – 1000 µL pipette tips

 
File Name Type Size Download
User Manual 972.08 KB

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