Vitamins A and E in Serum/Plasma - HPLC Basic Kit

Order No.: 34400-BK
Parameters:
Vitamin A (Retinol), Vitamin E (α-Tocopherol)

Short run time of less than 3 minutes
Optimized precipitation
Choice to use HPLC column at ambient temperature or at 35 °C

Assay validated according to IVDR (=> Declaration of Conformity)

Column temperature: 35 °C

Vitamin A (Retinol)

Vitamin E (α-Tocopherol)

Clinical relevance
This assay enables the simultaneous quantitative determination of vitamin A (retinol) and vitamin E (α-tocopherol) in human serum or plasma samples by HPLC.

It is intended to be used for determination of vitamin A and vitamin E levels where indicated
• for screening in patients with suspected vitamin A and/or E deficiency,
• for screening in patients with suspected vitamin A and/or E excess,
• for monitoring of patients under vitamin A and/or E supplementation.

 

Product advantages
• Short run time of less than 3 minutes
• Optimized precipitation
• Choice to use HPLC column at ambient temperature or at 35 °C
• Easy sample preparation
• No cooling of eluates – no cooled autosampler necessary
• No precolumn required
• No UHPLC needed: Short run time even with HPLC! – Also applicable to UHPLC systems

 

Assay characteristics
The fast and easy sample preparation can be performed with reaction vials or 96 deep well plates. The analytes are released from their transport protein or lipoproteins by precipitation. Subsequently, the substances are chromatographically separated in one isocratic HPLC run and quantified using a UV detector.

Detailed performance evaluation data can be found in Appendices II and III of the instruction manual.

 

Two types of kits available

Basic Kit
• Only sample preparation reagents and vials included
• No excess quantities of mobile phase and calibrators

34400-BK Basic Kit
For 200 sample preparations

34404 Internal Standard, 40 ml (1x)
34405 Precipitation Reagent, 40 ml (1x)
33005 Reaction Vials, 1.5 ml, amber coloured (light protection), 100 pcs. (2x)

Reagent Kit
• Mobile phase and calibrator included
 

34400 Reagent Kit
For 200 tests

34404 Internal Standard, 40 ml (1x)
34405 Precipitation Reagent, 40 ml (1x)
33005 Reaction Vials, 1.5 ml, amber coloured (light protection), 100 pcs. (2x)
34001 Mobile Phase, 1000 ml (1x)
34004 Serum Calibration Standard, 5 x 1 ml (lyoph.) (1x)
More Information
Method of Analysis HPLC
Sample Preparations 200
Please note The information listed here, including the sample preparation, is not sufficient for using the product. Please read the information provided in the instruction manual, which includes detailed information on limitations associated with the use of the product in line with its intended purpose. Detailed performance evaluation data can be found in Appendices II and III of the instruction manual.
Lower and Upper Limit of Quantitation

Serum:
Retinol: 0.02 – 5.0 mg/l
α-Tocopherol: 0.3 – 45 mg/l

Plasma:
Retinol: 0.01 – 5.0 mg/l
α-Tocopherol: 0.5 – 45 mg/l

Different systems might show different performance data.

Specimen Serum/Plasma
Sample Preparation

The information on the sample preparation presented here is not sufficient for use in the laboratory. For a detailed step by step description, please refer to the instruction manual.

Sample preparation with reaction vials:

  • Pipette 100 µL sample/calibrator/control into an amber coloured reaction vial.
  • Add 200 µL of Internal Standard, mix 30 s (vortex).
  • Centrifuge 10 min.
  • Add 200 µL of Precipitation Reagent, mix 30 s (vortex).
  • Centrifuge 2 min.
  • Transfer supernatant into a glass autosampler vial.
  • Inject 5 to 20 µL of the supernatant into the HPLC system.

For the sample preparation with 96 deep well plates, please refer to the instruction manual.

Run Time depending on column temperature: 2.9 min (35 °C) or 3.7 min (ambient)
Injection Volume 5 – 20 µl
Flow Rate 0.8 ml/min
Wavelengths

325 nm,
after 1.15 – 1.4 min switch to 295 nm

Additional Info For the analysis any isocratic HPLC system with UV detector is suitable. For simultaneous analysis of retinol and α-tocopherol, a detector with programmable wavelength switching is very useful as these analytes are detected at different wavelengths.

Parameters Vitamin A (Retinol), Vitamin E (α-Tocopherol)

Vitamins A and E in Serum/Plasma - HPLC Basic Kit (34400-BK)
For 200 sample preparations

Order No. Basic Kit components (Quantities in kit)
34404 Internal Standard, 40 ml (1x)
34405 Precipitation Reagent, 40 ml (1x)
33005 Reaction Vials, 1.5 ml, amber coloured (light protection), 100 pcs. (2x)

 

Order No. Required components (not included in the Basic Kit)
34300 HPLC Column, equilibrated, with test chromatogram, 1 pc.
34001 Mobile Phase, 1000 ml
34004 Serum Calibration Standard, 5 x 1 ml (lyoph.)
0036 Serum Control, Level I, 5 x 2 ml (lyoph.)
0037 Serum Control, Level II, 5 x 2 ml (lyoph.)
15012 PEEK-encased Prefilter, 0.5 µm, 1 pc.

 

Order No. General lab equipment (non-CE/IVD products)
33005 Reaction Vials, 1.5 ml, amber coloured (light protection), 100 pcs.
15070 Stainless Steel Prefilter Housing, 1 pc.
J0505 Micro-inserts for Autosampler Vials, clear glass, flat bottom, 6x31 mm, 0.4 ml, 100 pcs.
The following components are included in the kit:
The following products are not included in the kit but are required for the application of the method:
As a customer please login or register to gain full access.

Column temperature: 35 °C

Vitamin A (Retinol)

Vitamin E (α-Tocopherol)

Clinical relevance
This assay enables the simultaneous quantitative determination of vitamin A (retinol) and vitamin E (α-tocopherol) in human serum or plasma samples by HPLC.

It is intended to be used for determination of vitamin A and vitamin E levels where indicated
• for screening in patients with suspected vitamin A and/or E deficiency,
• for screening in patients with suspected vitamin A and/or E excess,
• for monitoring of patients under vitamin A and/or E supplementation.

 

Product advantages
• Short run time of less than 3 minutes
• Optimized precipitation
• Choice to use HPLC column at ambient temperature or at 35 °C
• Easy sample preparation
• No cooling of eluates – no cooled autosampler necessary
• No precolumn required
• No UHPLC needed: Short run time even with HPLC! – Also applicable to UHPLC systems

 

Assay characteristics
The fast and easy sample preparation can be performed with reaction vials or 96 deep well plates. The analytes are released from their transport protein or lipoproteins by precipitation. Subsequently, the substances are chromatographically separated in one isocratic HPLC run and quantified using a UV detector.

Detailed performance evaluation data can be found in Appendices II and III of the instruction manual.

 

Two types of kits available

Basic Kit
• Only sample preparation reagents and vials included
• No excess quantities of mobile phase and calibrators

34400-BK Basic Kit
For 200 sample preparations

34404 Internal Standard, 40 ml (1x)
34405 Precipitation Reagent, 40 ml (1x)
33005 Reaction Vials, 1.5 ml, amber coloured (light protection), 100 pcs. (2x)

Reagent Kit
• Mobile phase and calibrator included
 

34400 Reagent Kit
For 200 tests

34404 Internal Standard, 40 ml (1x)
34405 Precipitation Reagent, 40 ml (1x)
33005 Reaction Vials, 1.5 ml, amber coloured (light protection), 100 pcs. (2x)
34001 Mobile Phase, 1000 ml (1x)
34004 Serum Calibration Standard, 5 x 1 ml (lyoph.) (1x)
More Information
Method of Analysis HPLC
Sample Preparations 200
Please note The information listed here, including the sample preparation, is not sufficient for using the product. Please read the information provided in the instruction manual, which includes detailed information on limitations associated with the use of the product in line with its intended purpose. Detailed performance evaluation data can be found in Appendices II and III of the instruction manual.
Lower and Upper Limit of Quantitation

Serum:
Retinol: 0.02 – 5.0 mg/l
α-Tocopherol: 0.3 – 45 mg/l

Plasma:
Retinol: 0.01 – 5.0 mg/l
α-Tocopherol: 0.5 – 45 mg/l

Different systems might show different performance data.

Specimen Serum/Plasma
Sample Preparation

The information on the sample preparation presented here is not sufficient for use in the laboratory. For a detailed step by step description, please refer to the instruction manual.

Sample preparation with reaction vials:

  • Pipette 100 µL sample/calibrator/control into an amber coloured reaction vial.
  • Add 200 µL of Internal Standard, mix 30 s (vortex).
  • Centrifuge 10 min.
  • Add 200 µL of Precipitation Reagent, mix 30 s (vortex).
  • Centrifuge 2 min.
  • Transfer supernatant into a glass autosampler vial.
  • Inject 5 to 20 µL of the supernatant into the HPLC system.

For the sample preparation with 96 deep well plates, please refer to the instruction manual.

Run Time depending on column temperature: 2.9 min (35 °C) or 3.7 min (ambient)
Injection Volume 5 – 20 µl
Flow Rate 0.8 ml/min
Wavelengths

325 nm,
after 1.15 – 1.4 min switch to 295 nm

Additional Info For the analysis any isocratic HPLC system with UV detector is suitable. For simultaneous analysis of retinol and α-tocopherol, a detector with programmable wavelength switching is very useful as these analytes are detected at different wavelengths.

Parameters Vitamin A (Retinol), Vitamin E (α-Tocopherol)

Vitamins A and E in Serum/Plasma - HPLC Basic Kit (34400-BK)
For 200 sample preparations

Order No. Basic Kit components (Quantities in kit)
34404 Internal Standard, 40 ml (1x)
34405 Precipitation Reagent, 40 ml (1x)
33005 Reaction Vials, 1.5 ml, amber coloured (light protection), 100 pcs. (2x)

 

Order No. Required components (not included in the Basic Kit)
34300 HPLC Column, equilibrated, with test chromatogram, 1 pc.
34001 Mobile Phase, 1000 ml
34004 Serum Calibration Standard, 5 x 1 ml (lyoph.)
0036 Serum Control, Level I, 5 x 2 ml (lyoph.)
0037 Serum Control, Level II, 5 x 2 ml (lyoph.)
15012 PEEK-encased Prefilter, 0.5 µm, 1 pc.

 

Order No. General lab equipment (non-CE/IVD products)
33005 Reaction Vials, 1.5 ml, amber coloured (light protection), 100 pcs.
15070 Stainless Steel Prefilter Housing, 1 pc.
J0505 Micro-inserts for Autosampler Vials, clear glass, flat bottom, 6x31 mm, 0.4 ml, 100 pcs.
The following components are included in the kit:
The following products are not included in the kit but are required for the application of the method:
As a customer please login or register to gain full access.