Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 Apr 14;20(4):6611-25.
doi: 10.3390/molecules20046611.

Employing response surface methodology for the optimization of ultrasound assisted extraction of lutein and β-carotene from spinach

Ammar Altemimi  1   2 , David A Lightfoot  3 , Mary Kinsel  4 , Dennis G Watson  5
Affiliations

Employing response surface methodology for the optimization of ultrasound assisted extraction of lutein and β-carotene from spinach

Ammar Altemimi et al. Molecules. .

Abstract

The extraction of lutein and β-carotene from spinach (Spinacia oleracea L.) leaves is important to the dietary supplement industry. A Box-Behnken design and response surface methodology (RSM) were used to investigate the effect of process variables on the ultrasound-assisted extraction (UAE) of lutein and β-carotene from spinach. Three independent variables, extraction temperature (°C), extraction power (%) and extraction time (min) were studied. Thin-layer chromatography (TLC) followed by UV visualization and densitometry was used as a simple and rapid method for both identification and quantification of lutein and β-carotene during UAE. Methanol extracts of leaves from spinach and authentic standards of lutein and β-carotene were separated by normal-phase TLC with ethyl acetate-acetone (5:4 (v/v)) as the mobile phase. In this study, the combination of TLC, densitometry, and Box-Behnken with RSM methods were effective for the quantitative analysis of lutein and β-carotene from spinach extracts. The resulting quadratic polynomial models for optimizing lutein and β-carotene from spinach had high coefficients of determination of 0.96 and 0.94, respectively. The optimal UAE settings for output of lutein and β-carotene simultaneously from spinach extracts were an extraction temperature of 40 °C, extraction power of 40% (28 W/cm3) and extraction time of 16 min. The identity and purity of each TLC spot was measured using time-of-flight mass spectrometry. Therefore, UAE assisted extraction of carotenes from spinach can provide a source of lutein and β-carotene for the dietary supplement industry.

PubMed Disclaimer

Conflict of interest statement

None of the authors have any financial interest of conflict of interest with the results of this study.

Figures

Figure 1
Figure 1
Response surface model plot showing the effects of independent variables on Lutein content: panel A temperature and power; panel B temperature and time; and panel C power and time.
Figure 2
Figure 2
Response surface model plot showing the effects of independent variables on β-carotene content: panel A temperature and power; panel B temperature and time; and panel C power and time.
Figure 3
Figure 3
Mass spectra of spinach extract TLC spot LS of experimental condition (panel A) and TLC spot BS of experimental condition (panel C) excised and compared to lutein (panel B) and β-carotene (panel D) standards.
Figure 4
Figure 4
The chromatographic separation of lutein (L) and β-carotene (B): (A) real visible light image; (B) UV image at 254 nm; and (C) grey scale image by Quantity One software. Spots: 1 to 17 for spinach extracts; L1 and L2 for lutein controls; and B1 and B2 for β-carotene controls.
See this image and copyright information in PMC

References

    1. Gurib-Fakim A. Medicinal plants: Traditions of yesterday and drugs of tomorrow. Mol. Asp. Med. 2006;27:1–93. doi: 10.1016/j.mam.2005.07.008. - DOI - PubMed
    1. Nassr-Allah A.A., Aboul-Enein A.M., Aboul-Enein K.M., Lightfoot D.A., Cocchetto A., El-Shemy H.A. Anti-cancer and anti-oxidant activity of some Egyptian medicinal plants. J. Med. Plants Res. 2009;3:799–808.
    1. El-Shemy H.A., Aboul-Enein K.M., Lightfoot D.A. Predicting In Silico Which Mixtures of the Natural Products of Plants Might Most Effectively Kill Human Leukemia Cells? Evid. Based Complement. Altern. Med. 2013;2013:801501. doi: 10.1155/2013/801501. - DOI - PMC - PubMed
    1. Oliver J., Palou A. Chromatographic determination of carotenoids in foods. J. Chromatogr. A. 2000;881:543–555. doi: 10.1016/S0021-9673(00)00329-0. - DOI - PubMed
    1. West B.J., Deng S. Thin layer chromatography methods for rapid identity testing of Morinda citrifolia L. (Noni) fruit and leaf. Adv. J. Food Sci. Technol. 2010;2:298–302.

Publication types

MeSH terms

LinkOut - more resources