Vitamin A/E HPLC Kit - Labodia
Vitamin A/E HPLC Kit - Labodia Vitamin A/E HPLC Kit - Labodia
HPLC-Analytik / HPLC analytics Vitamin A/E Content 1. INTENDED USE ________________________________________________________ 15 2. INTRODUCTION _______________________________________________________ 15 3. PRINCIPLE OF THE TEST ________________________________________________ 15 4. MATERIAL SUPPLIED ___________________________________________________ 16 5. MATERIAL REQUIRED BUT NOT SUPPLIED _________________________________ 17 6. PREPARATION AND STORAGE OF REAGENTS ______________________________ 17 7. PRECAUTIONS ________________________________________________________ 17 8. SPECIMEN COLLECTION AND PREPARATION _______________________________ 17 9. ASSAY PROCEDURE ____________________________________________________ 18 Procedural notes ________________________________________________________ 18 Sample and standard preparation __________________________________________ 18 Chromatographic conditions ______________________________________________ 19 10. TREATMENT OF THE COLUMN ___________________________________________ 19 11. RESULTS _____________________________________________________________ 19 Calculation ____________________________________________________________ 19 Typical chromatogram ___________________________________________________ 20 12. LIMITATIONS __________________________________________________________ 20 13. QUALITY CONTROL ____________________________________________________ 20 14. PERFORMANCE CHARACTERISTICS ______________________________________ 21 Precision and reproducibility ______________________________________________ 21 Linearity ______________________________________________________________ 21 Detection limit _________________________________________________________ 21 Recovery ______________________________________________________________ 21 15. DISPOSAL ____________________________________________________________ 21 16. TROUBLESHOOTING ___________________________________________________ 22 17. REFERENCES __________________________________________________________ 23 18. GENERAL NOTES ON THE TEST AND TEST PROCEDURE ______________________ 23 14
HPLC-Analytik / HPLC analytics Vitamin A/E 1. INTENDED USE The Immundiagnostik Assay is intended for the quantitative determination of Vitamin A/E in serum and plasma. This Assay is designed for in vitro diagnostic use only. 2. INTRODUCTION Vitamin A (Retinol) and Vitamin E (Tocopherol) are fat-soluble vitamins, which could be stored for longer periods in the adipose tissue. Both, lack and excess could express in complaints. Vitamin A is essential for the visual process and recovers the skin and mucosa. A lack of Vitamin A will reduce the visual power up to total blindness. An excess of Vitamin A could cause headache, damage of the skin, liver disease, painful alteration in the skeleton or foetal damage. Vitamin E (Tocopherol) is an antioxidant and protects unsaturated fatty acids against oxidation. It also protects the cells of the body by catching radicals. A lack of vitamin E in animal experiments demonstrates diseases of muscle, nervous system, heart, liver and reproduction system. These symptoms were not observed in humans. Vitamin E can be stored in the adipose tissue in large amounts. A lack can be caused by a malfunction in digestion or resorption of fatty acids. 3. PRINCIPLE OF THE TEST The first step in the determination of Vitamin A and E includes the sample preparation. In the first step an internal standard solution is added. During the precipitation higher molecular substances are removed. After centrifugation the supernatant is used for injection into the HPLC system. The separation via HPLC follows an isocratic method at 30°C using a „reversed phase“ column; one run lasts 15 minutes. The detection is performed by an UV detector at two different wavelengths (Vitamin A: 325 nm; Vitamin E: 300 nm). The quantification is performed with the delivered standard solution; the concentration is calculated via integration of the peak areas in the internal standard calibration mode. 15
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<strong>HPLC</strong>-Analytik / <strong>HPLC</strong> analytics <strong>Vitamin</strong> A/E<br />
Content<br />
1. INTENDED USE ________________________________________________________ 15<br />
2. INTRODUCTION _______________________________________________________ 15<br />
3. PRINCIPLE OF THE TEST ________________________________________________ 15<br />
4. MATERIAL SUPPLIED ___________________________________________________ 16<br />
5. MATERIAL REQUIRED BUT NOT SUPPLIED _________________________________ 17<br />
6. PREPARATION AND STORAGE OF REAGENTS ______________________________ 17<br />
7. PRECAUTIONS ________________________________________________________ 17<br />
8. SPECIMEN COLLECTION AND PREPARATION _______________________________ 17<br />
9. ASSAY PROCEDURE ____________________________________________________ 18<br />
Procedural notes ________________________________________________________ 18<br />
Sample and standard preparation __________________________________________ 18<br />
Chromatographic conditions ______________________________________________ 19<br />
10. TREATMENT OF THE COLUMN ___________________________________________ 19<br />
11. RESULTS _____________________________________________________________ 19<br />
Calculation ____________________________________________________________ 19<br />
Typical chromatogram ___________________________________________________ 20<br />
12. LIMITATIONS __________________________________________________________ 20<br />
13. QUALITY CONTROL ____________________________________________________ 20<br />
14. PERFORMANCE CHARACTERISTICS ______________________________________ 21<br />
Precision and reproducibility ______________________________________________ 21<br />
Linearity ______________________________________________________________ 21<br />
Detection limit _________________________________________________________ 21<br />
Recovery ______________________________________________________________ 21<br />
15. DISPOSAL ____________________________________________________________ 21<br />
16. TROUBLESHOOTING ___________________________________________________ 22<br />
17. REFERENCES __________________________________________________________ 23<br />
18. GENERAL NOTES ON THE TEST AND TEST PROCEDURE ______________________ 23<br />
14
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