How to Measure the Serum Lycopene and Beta-Carotene Simultaneously By Using High Performance Liquid

Introduction

Lycopene is a natural pigment of tomato and a strong anti oxidant, which activity had a synergistic effect with Vitamin E. Beta-carotene is one of the carote-noid provitamins A. The both substances have the anti cancer effects, which have been approved in many studies by researchers and scientists. This article is mainly focusing on the measurement of lycopene and beta-carotene using high performance liquid chromatography.

Sample preparation

Materials

N-hexane, methanol, ethanol, butylated hydroxyl toluene, acetonitrile and tetrahydrofuran, purity degree HPLC, BDH, standard lycopene and standard beta-carotene.

Apparatuses

HPLC apparatus is equipped with an ultraviolet detector, Novopack C18 and printer.

Standard solutions preparation

Prepare various standards of lycopene and beta-cartene in mobile phase.

Procedures

Add 1000µL of ethanol to 500µL of serum sample and mix them for 20 seconds with electrical mixer in order to precipitate proteins. And then add 1000µL of hexane to it, mixing for 60 seconds. Transferring the upper phase to an injection vial. Perform the extraction stage again. Then, evaportate the organic phase of hecane at 40ºC under flow of nitrogen gas and add the obtained precipitant to mobile phase, which contains diethyl ether in order to change its volume to 500µL. And then inject this sample into column.

Quality test

1. Range of linearity: standard curve were prepared for lycopene with concentrations of 6, 15, 30, 60, 120 and 150 micrograms per deciliter and for beta-carotene should be with concentration of 10, 25, 50, 100, 200 and 250 micrograms per deciliter.
2. Detection limit: the sequential attenuating the lowest concentration, which is used for preparing standard curve, can determine the detection limit.
3. Recovery percent: add 20µL of standard lycopene and beta-carotene to 500µL of serum sample. And the two substances should be prepared similar to ordinary sample. Then, injecting 20µL of it into the column and calculate recovery percent.
4. Precision: the repeated testing of a unit mixed serum sample in a day and sequential days can be used to determine repeatability of test, intra- and inter-assay variations.

These are some basic but useful information and procedures to detect lycopene and beta-carotene simultaneously by using high-quality high performance liquid chromatography services. And these services can be utilized in many aspects.