Optimization of Ultrasound-Assisted Extraction Conditions Followed by Solid Phase Extraction Fractionation from Orthosiphon stamineus Benth (Lamiace) Leaves for Antiproliferative Effect on Prostate Cancer Cells
Abstract:Primarily, optimization of ultrasonic-assisted extraction (UAE) conditions of Orthospihon stamineus was evaluated and verified using a central composite design (CCD) based on three factors including extraction time (minutes), ultrasound amplitude (A), and solvent concentration (%). The response surface methodology (RSM) was performed to develop an extraction method with maximum yield and high rosmarinic acid content. The optimal UAE conditions were as follows: extraction time 21 min, ultrasound amplitudes 62 A… Show more
“…A similar trend was found in several previous studies reported by Syakfanaya et al [39]. A study conducted by Suhaimi et al [40] reported that UAE of more than 20 min caused a decrease of phenolic compound yields.…”
Section: Extraction Timesupporting
confidence: 90%
“…A study conducted by Suhaimi et al . [ 40 ] reported that UAE of more than 20 min caused a decrease of phenolic compound yields. Figure 3.…”
Section: Resultsmentioning
confidence: 99%
“…The lack-of-fit value of a good model must be insignificant. This means that the failure of the model to represent the data is not significant, so the model is appropriate to predict the responses [ 40 ]. A p -value < 0.05 in the model indicates a significant effect of the variable on the response and our results showed that there are interactions between the variables.…”
In this study, an environmentally friendly extraction method for flavonoid compound from
Ixora javanica
, as a new raw material candidate for herbal medicine and cosmetics, was developed. The objectives of the present work were to provide recommendations for the optimal extraction conditions and to investigate the effects of any extraction parameters on flavonoid yields from the
I. javanica
flower. The extraction process was performed using deep eutectic solvent (DES) (choline chloride and propylene glycol at molar ratio of 1 : 1) and the ultrasound-assisted extraction method. Both single-factor and response surface analyses using three-level and three-factor Box Behnken designs were conducted to obtain the optimum flavonoid concentrations. The results showed that the optimum extraction conditions for total flavonoids featured an extraction time of 40 min, 25% water content in DES and a solid-to-liquid ratio of 1 : 25 g ml
−1
. An extract obtained under optimum extraction conditions showed higher total flavonoid yields than an ethanolic extract which was used for comparison. Scanning electron microscope images demonstrated that both of the solvents also showed different effects on the outer surface of the
I. javanica
flower during the extraction process. In summary, our work succeeded in determining the optimum conditions for total flavonoids in the
I. javanica
flower using a green extraction method.
“…A similar trend was found in several previous studies reported by Syakfanaya et al [39]. A study conducted by Suhaimi et al [40] reported that UAE of more than 20 min caused a decrease of phenolic compound yields.…”
Section: Extraction Timesupporting
confidence: 90%
“…A study conducted by Suhaimi et al . [ 40 ] reported that UAE of more than 20 min caused a decrease of phenolic compound yields. Figure 3.…”
Section: Resultsmentioning
confidence: 99%
“…The lack-of-fit value of a good model must be insignificant. This means that the failure of the model to represent the data is not significant, so the model is appropriate to predict the responses [ 40 ]. A p -value < 0.05 in the model indicates a significant effect of the variable on the response and our results showed that there are interactions between the variables.…”
In this study, an environmentally friendly extraction method for flavonoid compound from
Ixora javanica
, as a new raw material candidate for herbal medicine and cosmetics, was developed. The objectives of the present work were to provide recommendations for the optimal extraction conditions and to investigate the effects of any extraction parameters on flavonoid yields from the
I. javanica
flower. The extraction process was performed using deep eutectic solvent (DES) (choline chloride and propylene glycol at molar ratio of 1 : 1) and the ultrasound-assisted extraction method. Both single-factor and response surface analyses using three-level and three-factor Box Behnken designs were conducted to obtain the optimum flavonoid concentrations. The results showed that the optimum extraction conditions for total flavonoids featured an extraction time of 40 min, 25% water content in DES and a solid-to-liquid ratio of 1 : 25 g ml
−1
. An extract obtained under optimum extraction conditions showed higher total flavonoid yields than an ethanolic extract which was used for comparison. Scanning electron microscope images demonstrated that both of the solvents also showed different effects on the outer surface of the
I. javanica
flower during the extraction process. In summary, our work succeeded in determining the optimum conditions for total flavonoids in the
I. javanica
flower using a green extraction method.
“…At investigation of single-factors in uence on emodin yield, the overall trend is similar: With the increase of variables, it rst increases to a certain vertex and then decreases. For in uence of ultrasonic time, on ultrasonic extraction of rosmarinic acid from Orthosiphon stamineus Benth leaves, the similar tendency was found [47]. In general, the ultrasonic wave needs time to crush the cell walls of R. o cinale and extract the cell contents.…”
Background
Streptococcus suis is a zoonotic pathogen that causes serious systemic infections in pigs and humans. It is a great threat to pig breeding and public health safety. The formation of biofilm was one of the main reasons that it’s difficult to cure. Rhubarb water extract can inhibit the formation of biofilm of Streptococcus suis, but the main functional ingredient was not clear. And what’s the potential mechanism of emodin intervene the biofilm formation? Importantly, the gene expression of luxS of Streptococcus suis was significantly decreased with emodin treatment, if there is a possibility that emodin could combine with LuxS protein which in turn inhibit the formation of biofilm?
Methods
Optimization of green ultrasonic extraction of emodin from R. officinale by Response Surface Methodology. The activity of antibacterial was evaluated by MIC assay. And the ability that intervene biofilm formation was evaluated through Crystal violet staining and SEM. The combination mode between emodin and LuxS protein was identified through molecular docking.
Results
The optimum extraction conditions (ethanol concentration of 80%, extraction time of 21 min, and liquid to solid ratio of 12.5:1 mL/g) were determined through the interaction analysis of different influencing factors and model verification experiment. The minimal inhibitory concentration (MIC) of RUEP against S.suis was 15.625 µg·mL-1, and sub-inhibitory concentrations of RUEP (1/2, 1/4, 1/8 and 1/16 MIC) significantly inhibited the biofilm formation of S. suis. Moreover, scanning electron microscopy analysis confirmed that RUEP can damage the layered colony community structure in biofilm and effectively intervene in the biofilm formation of Streptococcus suis. Through molecular docking analyzed, the inhibition of biofilm formation could be realized through the formation of hydrogen bond with residues His14 and Glu 60, π-π staking with His61、Phe80 and hydrophobic interaction with Cys127.
Conclusion
Through our study, we found emodin have good antibacterial and antibiofilm formation properties and the potential mechanism of emodin intervention on biofilm formation of Streptococcus suis was inferred.
“…It means that the sinensetin is major flavonoid in studied extract genotypes. On the other hand, it was reported that sinensetin content will decrease when extraction was performed using polar solvent such as ethanol 70% 34 . This result indicated that the genotype A15 and A11 could be developed through a breeding program to get the high flavonoid and sinensetin contents of O. aristatus varieties.…”
Orthosiphon aristatus (Blume) Miq. of the Lamiaceae family, called as kumis kucing in Indonesia, is a valuable medicinal plant for their pharmacological properties. The present study comprised of fifteen genotypes of O. aristatus was undertaken to evaluate the genotypes based on phytochemical content and pharmacological activities of leaves ethanol extract. Chemometric analysis (correlation and principal component analysis) was also used to investigate the genetic variability based on phytochemical content and pharmacological activities of O. aristatus genotypes. Results of phytochemical characterization showed that total phenolic ranged from 1.48 to 36.08 (maximum in A15) mg GAE/g DW, total flavonoid ranged from 0.10 to 3.07 (maximum in A15) mg QE/g DW, sinensetin ranged from 0.36 to 4.02 (maximum in A11) mg/g DW, and rosmarinic acid ranged 0.06 to 7.25 (maximum in A7) mg/g DW. Antioxidant activity was tested using DPPH and FRAP assay. Antioxidant results showed that DPPH ranged from 1.68 to 15.55 (maximum in A15) μmol TE/g DW and FRAP ranged from 0.07 to 1.60 (maximum in A1 and A7) μmol TE/g DW. The genotype A8 showed the highest cytotoxic activities against HeLa (66.25%) and MCF-7 (61.79%) cell lines. Maximum α-glucosidase inhibitory activity was recorded in genotype A2 with the value of 62.84%. The genotypes A1, A2, A7, A11, and A15 were identified as superior based on their phytochemicals content and pharmacological activities coupled with chemometric analysis. This finding is important for breeding studies and also the pharmaceutical perspective of O. aristatus.
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