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. 2024 Jun 1;29(11):2610.
doi: 10.3390/molecules29112610.

Optimization of Flavonoid Extraction from Abelmoschus manihot Flowers Using Ultrasonic Techniques: Predictive Modeling through Response Surface Methodology and Deep Neural Network and Biological Activity Assessment

Zenghong Sun  1 , Wenhui Liu  2 , Sha Zhang  1 , Shuge Tian  1 , Ainiwaer Aikemu  3
Affiliations

Optimization of Flavonoid Extraction from Abelmoschus manihot Flowers Using Ultrasonic Techniques: Predictive Modeling through Response Surface Methodology and Deep Neural Network and Biological Activity Assessment

Zenghong Sun et al. Molecules. .

Abstract

Understanding the optimal extraction methods for flavonoids from Abelmoschus manihot flowers (AMF) is crucial for unlocking their potential benefits. This study aimed to optimize the efficiency of flavonoid extraction from AMF. After comparing extraction methods, the ultrasonic cell crusher demonstrated superior performance over conventional techniques. Four key factors-solid-to-liquid ratio (1:10 to 1:50 g·mL-1), ethanol concentration (55% to 95%), ultrasonic time (10 to 50 min), and ultrasonic power (5% to 25% of 900 W)-were investigated and normalized using the entropy weight method. This led to a comprehensive evaluation (CE). Optimization of extraction conditions for the ultrasonic cell crusher was achieved through response surface methodology and a deep neural network model, resulting in optimal parameters: ethanol volume fraction of 66%, solid-to-liquid ratio of 1:21 g/mL, extraction efficiency of 9%, and extraction duration of 35 min, yielding a CE value of 23.14 (RSD < 1%). Additionally, the inhibitory effects of the optimized extracts against Streptococcus mutans (S. mutans) were assessed. The results revealed that AMF extract (AMFE) exhibits inhibitory effects on S. mutans, with concomitant inhibition of sucrase and lactate dehydrogenase (LDH). The MIC of AMFE against planktonic S. mutans is 3 mg/mL, with an MBC of 6 mg/mL. Within the concentration range of 1/8 MIC to 2 MIC of AMFE, the activities of sucrase and LDH decreased by 318.934 U/mg prot and 61.844 U/mg prot, respectively. The antioxidant activity of AMFE was assessed using the potassium ferricyanide reduction and phosphomolybdenum methods. Additionally, the effect of AMFE on DPPH, ABTS, and ·OH free radical scavenging abilities was determined. The concentrations at which AMFE exhibited over 90% scavenging rate for ABTS and DPPH free radicals were found to be 0.125 mg/mL and 2 mg/mL, respectively.

Keywords: Abelmoschus manihot flower; Streptococcus mutans (S. mutans); antioxidant activity; dental caries; green extraction; ultrasonic cell crusher.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
The OM and SEM images of AMF powder. (A–C) Maceration extraction, (D–F) reflux extraction, (G–I) ultrasonic extraction.
Figure 2
Figure 2
Effect of various factors on the content of active components from AMF. (A) ethanol concentration, (B) solid-to-liquid ratio, (C) extraction time, (D) ultrasonic power.
Figure 3
Figure 3
Contours of the influence of different factor interactions on CE. Note: The closer the color in the figure is to red, the higher the CE value, with the dots being the maximum value.
Figure 4
Figure 4
Three-dimensional response surface diagrams of the influence of different factor interactions on CE. A: ethanol concentration (%), B: liquid-to-solid ratio (g/mL), C: power (%), D: time. Note: The closer the color in the figure is to red, the higher the CE value, with the dots being the maximum value.
Figure 5
Figure 5
DNN model topology.
Figure 6
Figure 6
Training the DNN model using the LM algorithm (A) and the GA algorithm (B).
Figure 7
Figure 7
Correlation coefficient index for model construction of the DNN; Fitting equation for training set (A); Fitting equation for validation set (B); Fitting equation for testing set (C); Fitting equation for all sets (D).
Figure 8
Figure 8
HPLC (A) and TIC (B) plots of AMF.
Figure 9
Figure 9
Scan ES- and daughters of ES- of four flavonoids. (A) Rutin, (B) Hyp, (C) Iso, (D) Que.
Figure 10
Figure 10
(A–C) Inhibition zone against S. mutans. (Note: N represents negative control, Y represents positive control, A1–A3, B1–B3, and C1–C3 represent 50, 25, and 12.5 mg/mL AMFE solution, respectively). (D) Inhibitory effect on sucrase. (E) Inhibitory effect on LDH. (Note: a, b represents a statistically significant difference compared to a negative control and a positive control at p < 0.001 respectively, c indicates a statistically significant difference between any two experimental groups at p < 0.001).
Figure 11
Figure 11
Antioxidant capacity of the AMF extracted by optimized methods; total restoration capability (A), total antioxidant capacity (B), ABTS radical scavenging capability (C), ·OH radical scavenging capacity (D), and DPPH radical scavenging capability (E).
See this image and copyright information in PMC

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