HPLC Method for Analysis of Lycopene, beta-Carotene on Lipak Column by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of Lycopene, beta-Carotene.

Lycopene is a carotenoid hydrocarbon with C₄₀H₅₆ molecular formula. It is found primarily in tomatoes, watermelon, grapefruits, and red guavas. Gac has the highest concentration of lycopene at 2~6 per gram. Lycopene in non-toxic, but can be a rare allergen. You can find detailed UV spectra of Lycopene and information about its various lambda maxima by visiting the following link.

beta-Carotene, also written as β-Carotene, is a pigment with the chemical formula C₄₀H₅₆. It is found primarily in fungi, plants, and fruit. Especially high amounts of it are found in carrots, pumpkins, spinach, and sweet potatoes. In dietary contexts, it is used as a pro-vitamin, as it converts to retinol in the body. In fact, it is the worldwide primary dietary source of provitamin A and is the best known provitamin a carotenoid. You can find detailed UV spectra of beta-Carotene and information about its various lambda maxima by visiting the following link.

*Lycopene  and β – Carotene  Extraction from Tomato

Weigh 5.0 g of homogenized tomato sample into a centrifuge tube.

Add 10.0 mL of hexane/acetone (3:1, v/v).

Mix the sample using an ika ultra turrax tube drive until  or an equivalent tube homogenizer/mixer until the sample is thoroughly homogenized and extracted.

Centrifuge for 5–10 minutes to separate the solid matrix from the organic extract.

Carefully transfer the clear organic supernatant to a clean tube.

Protect the extract from direct light throughout the procedure.

If necessary, dilute the extract with the extraction solvent to obtain a suitable concentration for HPLC analysis.

Filter the final solution through a 0.45 µm PTFE filter before injection.

Inject 2 µL of the filtered extract into the HPLC system.

Lycopene, beta-Carotene can be retained and analyzed using the Lipak stationary phase column. The analysis utilizes an isocratic method with a simple mobile phase consisting of water and acetonitrile (MeCN) and ethanol. Detection is performed using Vis.

Condition

Description

*Lycopene and β – Carotene Extraction from Tomato
Weigh 5.0 g of homogenized tomato sample into a centrifuge tube.
Add 10.0 mL of hexane/acetone (3:1, v/v).
Mix the sample using an ika ultra turrax tube drive until or an equivalent tube homogenizer/mixer until the sample is thoroughly homogenized and extracted.
Centrifuge for 5–10 minutes to separate the solid matrix from the organic extract.
Carefully transfer the clear organic supernatant to a clean tube.
Protect the extract from direct light throughout the procedure.
If necessary, dilute the extract with the extraction solvent to obtain a suitable concentration for HPLC analysis.
Filter the final solution through a 0.45 µm PTFE filter before injection.
Inject 2 µL of the filtered extract into the HPLC system.