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. 2022 Jan 26;27(3):820.
doi: 10.3390/molecules27030820.

A Green Extraction Method to Achieve the Highest Yield of Limonin and Hesperidin from Lime Peel Powder (Citrus aurantifolia)

Pakkapong Phucharoenrak  1 , Chawanphat Muangnoi  2 , Dunyaporn Trachootham  2
Affiliations

A Green Extraction Method to Achieve the Highest Yield of Limonin and Hesperidin from Lime Peel Powder (Citrus aurantifolia)

Pakkapong Phucharoenrak et al. Molecules. .

Abstract

Green extraction is aimed at reducing energy consumption by using renewable plant sources and environmentally friendly bio-solvents. Lime (Citrus aurantifolia) is a rich source of flavonoids (e.g., hesperidin) and limonoids (e.g., limonin). Manufacturing of lime products (e.g., lime juice) yields a considerable amount of lime peel as food waste that should be comprehensively exploited. The aim of this study was to develop a green and simple extraction method to acquire the highest yield of both limonin and hesperidin from the lime peel. The study method included ethanolic-aqueous extraction and variable factors, i.e., ethanol concentrations, pH values of solvent, and extraction temperature. The response surface methodology was used to optimize extraction conditions. The concentrations of limonin and hesperidin were determined by using UHPLC-MS/MS. Results showed that the yields of limonin and hesperidin significantly depended on ethanol concentrations and extraction temperature, while pH value had the least effect. The optimal extraction condition with the highest amounts of limonin and hesperidin was 80% ethanol at pH 7, 50 °C, which yields 2.072 and 3.353 mg/g of limonin and hesperidin, respectively. This study illustrates a green extraction process using food waste, e.g., lime peel, as an energy-saving source and ethanol as a bio-solvent to achieve the highest amount of double bioactive compounds.

Keywords: LC-MS/MS; citrus; food waste; green extraction; hesperidin; lime; limonin.

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Conflict of interest statement

The authors declare no conflict of interest or personal relationships with other people or organizations that can inappropriately influence our work.

Figures

Figure 1
Figure 1
Reference compounds analyzed by UHPLC-MS/MS. Precursor MS spectra of limonin (A) and hesperidin (B). Chromatograms for quantified (upper) and confirmed (lower) product ions of limonin (C) and hesperidin (D). Retention times are shown as a red number above the quantified peaks. Average calibration curves of limonin (E) and hesperidin (F) generated by the linear plots between areas under the curves of the quantitative product ions and the concentrations of standard solutions. R2 values were obtained from linear regression.
Figure 2
Figure 2
Effect of pH on extraction efficiency. Limonin (A) and hesperidin (B) concentrations in the ethanolic-aqueous extracts of lime peel after extraction with 60% (●), 70% (■), 80% (▲), and 100% (▼) ethanol in water with the specified pH values at 50 °C for 100 min. The statistical differences were separately analyzed for limonin and hesperidin by using one-way ANOVA followed by Tukey’s multiple comparisons test. * , **, *** and **** mean p < 0.05, <0.01, <0.001 and <0.0001, respectively, compared with the concentrations of limonin or hesperidin at pH 7 of each specified condition.
Figure 3
Figure 3
Effect of temperature on extraction efficiency. Limonin (A) and hesperidin (B) concentrations in the ethanolic-aqueous extracts of lime peel after extraction with 60% (●), 70% (■), 80% (▲), and 100% (▼) ethanol in water with the specified temperatures, at pH 7 for 100 min. The statistical differences were separately analyzed for limonin and hesperidin by using one-way ANOVA followed by Tukey’s multiple comparisons test. * , **, *** and **** mean p < 0.05, <0.01, <0.001 and <0.0001, respectively, compared with the concentrations of limonin or hesperidin at 50 °C of each specified condition.
Figure 4
Figure 4
Relationship between extraction factors. Three-dimensional (3D) response surface methodology (RSM) diagrams show concentrations of limonin (A–C) and hesperidin (D–F) in the ethanolic-aqueous extracts of lime peel as the function of ethanol concentration, extraction temperature, and pH value of extraction solvent. Red, yellow, green and blues areas depict the highest to lowest concentrations of limonin and hesperidin.
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