Optimization of Triterpenoid Extracted from Vietnamese Ganoderma lucidum via Supercritical Extraction Method and Biological Tests

Supercritical CO(2) was utilised to extract Chinese star anise seed oil (CSASO), and a three-level Box-Behnken factorial design from response surface methodology was applied to optimise the extraction conditions, including pressure, temperature and amount of modifier (ethanol). The compositional analysis of fatty acids in CSASO was performed by HPLC with fluorescence detection using 2-(11H-benzo[a]carbazol-11-yl)-ethyl-4-methylbenzenesulfonate (BCETS) as labelling reagent. Identification was carried out by online atmospheric chemical ionisation-mass spectrometry. The optimum extraction conditions were as follows: extraction pressure, 27.72 MPa, extraction temperature, 46.22 degrees C, and amount of modifier, 8.58 vol.%. The experimental result showed that the maximum extraction yield was 25.31 +/- 0.22% (w/w) under the conditions proposed. The compositional analysis indicated that CSASO mainly contained C18:2, C18:1, C18:3, C20:4, C16, C18 and C20 fatty acids. In this study, a fast, simple and high-efficiency supercritical technique for extracting oil from Chinese star anise seed was developed. Simultaneous determination of fatty acids in CSASO using BCETS as the labelling reagent with HPLC fluorescence detection and online mass spectroscopy identification has been successfully achieved.

Background/Objective Vitis vinifera leaves harvested from the developing canes are used in Balkan cuisines. The objective was to determine the chemical composition of the essential oil and supercritical carbon dioxide (SC CO2) extract of V. vinifera leaves harvested after the flowering of the new vine shoots. Methods The V. vinifera leaf oils were obtained by hydrodistillation and SC CO2 extraction. The application of different SC CO2 extraction parameters, determined by the Box–Behnken design, influenced the extraction yield and the chemical composition of the SC CO2 extracts. The optimal extraction conditions were set for the extraction yield and eucalyptol amount. The essential oil and SC CO2 extracts obtained were analyzed by gas chromatography–mass spectrometry. Results The most abundant compounds present in the essential oil obtained by hydrodistillation were: phytol (11.3%), α-cadinol (8.3%), trans-calamenene (8.2%), hexahydrofarnesyl acetone (7.0%), and τ-cadinol (6.6%). The extracts obtained by SC CO2 extraction consisted of twenty five compounds among which the most abundant were: phytol, squalene, eucalyptol, camphene and hexadecan-1-ol. Over the ranges of SC CO2 process parameters studied, those of pressure and extraction time had significant effect on the extraction yield and eucalyptol amount. Conclusion The results obtained determined the compounds present in V. vinifera leaves essential oil and SC CO2 extracts which contribute to the flavor and aroma of V. vinifera leaves.

In this experiment, the Box–Behnken design (BBD) of the response surface methodology (RSM) was used to optimize the ultrasound-assisted extraction variables (liquid-to-solid ratio, extraction temperature, and time) to obtain the maximum yield of physcion from the methanol extract of S. occidentalis (aerial parts). The analysis of physcion in the extracts obtained by using the optimized extraction condition was carried out in a gradient system by the HPLC-UV method with 0.5% formic acid in ultra-pure water (Solvent A) and acetonitrile (Solvent B) in different ratios as the mobile phase. The optimal extraction conditions for the maximum physcion extraction were found as: a liquid-to-solid ratio of 20.16 mL/g, extraction temperature of 52.2 °C, and extraction time of 46.6 min. Under these optimal ultrasonic extraction conditions, the experimental yield (% w/w of the dried extract) of the physcion was found to be 2.43%, which agreed closely with the predicted value (2.41). The experimental value was consistent with the value predicted by the RSM model, thus validating the fitness of the employed model and the success of the RSM in optimizing the extraction conditions. In future, this optimized ultrasonic extraction condition can be used in the maximum extraction of physcion from marketed herbal supplements containing S. occidentalis as well as other Senna species.

Isoflavones from Chinese soybean cheese were extracted with aqueous ethanol. Single-factor experiment design was employed to optimize the solid-to-liquid ratio (in grams per milliliter), ethanol concentration (in percent), extraction time (in hours), and extraction temperature (in degrees Celsius). The solid-to-liquid ratio was 1.5:10 (g/mL), and ethanol concentration (50 %–90 %), extraction time (2–3 h), extraction temperature (50–70 °C) were used for further optimization of extraction conditions. The optimal conditions for the ultrasound-assisted extraction of total isoflavones from Chinese soybean cheese were determined using response surface methodology (RSM) by Box–Behnken design. Three variables “ethanol concentration, extraction time and extraction temperature” were regarded as factors in the optimization study. The optimal conditions for total isoflavone extraction in Chinese soybean cheese were: ethanol concentration of 65.43 %, extraction temperature of 65.38 °C, and extraction time of 2.51 h. The verification experimental OD value was 0.534, which agreed with the predicted value, thus indicating suitability of the model employed and the suitability of RSM in optimizing the extraction conditions.

In this study, ultrasound-assisted extraction conditions were optimized to maximize the yields of sennoside A, sennoside B, aloe-emodin, emodin, and chrysophanol from S. alexandrina (aerial parts). The three UAE factors, extraction temperature (S1), extraction time (S2), and liquid to solid ratio (S3), were optimized using response surface methodology (RSM). A Box–Behnken design was used for experimental design and phytoconstituent analysis was performed using high-performance liquid chromatography-UV. The optimal extraction conditions were found to be a 64.2 °C extraction temperature, 52.1 min extraction time, and 25.2 mL/g liquid to solid ratio. The experimental values of sennoside A, sennoside B, aloe-emodin, emodin, and chrysophanol (2.237, 12.792, 2.457, 0.261, and 1.529%, respectively) agreed with those predicted (2.152, 12.031, 2.331, 0.214, and 1.411%, respectively) by RSM models, thus demonstrating the appropriateness of the model used and the accomplishment of RSM in optimizing the extraction conditions. Excellent antioxidant properties were exhibited by S. alexandrina methanol extract obtained using the optimized extraction conditions with a DPPH assay (IC50 = 59.7 ± 1.93, µg/mL) and ABTS method (47.2 ± 1.40, µg/mL) compared to standard ascorbic acid.

Cratoxylum formosum ssp. pruniflorum (Kurz.) Gogel (Guttiferae), called kuding tea, is widely distributed in Southeast Asia. In this study, the constituents and biological activity of C. formosum ssp. pruniflorum were investigated. Extract of its leaves, roots and stems showed antioxidant and α-glucosidase inhibitory activity. Interestingly, comparison of the metabolite profiles of leaves, roots and stems of C. formosum ssp. pruniflorum by LC-MS analysis showed a great difference between the roots and leaves, whereas the roots and stems were quite similar. Purification of the roots and leaves of C. formosum ssp. pruniflorum through various chromatographic techniques resulted in the isolation of 25 compounds. The structures of isolated compounds were elucidated on the basis of spectroscopic analysis as 18 xanthones, 5 flavonoids, a benzophenone and a phenolic compound. Among them, a xanthone (16) and a benzophenone (19) were first reported from nature. Evaluation of biological activity revealed that xanthones had a potent α-glucosidase inhibitory activity, while flavonoids were responsible for the antioxidant activity. To maximize the biological activity, yield and total phenolic content of C. formosum ssp. pruniflorum, extraction conditions such as extraction solvent, time and temperature were optimized using response surface methodology with Box–Behnken Design (BBD). Regression analysis showed a good fit of the experimental data, and the optimal condition was obtained as MeOH concentration in EtOAc, 88.1%; extraction time, 6.02 h; and extraction temperature 60.0 °C. α-Glucosidase inhibitory activity, yield and total phenolic content under the optimal condition were found to be 72.2% inhibition, 10.3% and 163.9 mg GAE/g extract, respectively. These results provide useful information about C. formosum ssp. pruniflorum as functional foods for oxidative stress–related metabolic diseases.

Optimization of ultrasonic-assisted extraction of phenolic compounds from bovine pennyroyal (Phlomidoschema parviflorum) leaves using response surface methodology

최적의 추출조건을 찾는데 매우 유용한 방법인 반응 표면 분석법(RSM, response surface methodology)을 사용하여 새우껍질로부터 astaxanthin 추출조건을 최적화하였다. 추출조건은 용매 에탄올과 추출물질의 비율, 추출온도(<TEX>$^{\circ}C$</TEX>), 추출시간(min)의 세가지 독립변수를 설정하여 BBD (Box-Behnken design) 방법을 이용하였다. 이 BBD 모델링은 0.9218의 <TEX>$R^2{_{adj}}$</TEX>값과 0.0003의 확률 값 p 값으로 회귀 모델에 대한 신뢰도를 입증하였다. RSM 분석을 통해 찾아낸 새우껍질로부터 astaxanthin의 최적 추출조건은 에탄올 용매비 1:29.7, 추출온도 <TEX>$49.5^{\circ}C$</TEX>, 추출시간 59.9 분이고, 이 때 astaxanthin 추출량은 <TEX>$17.80{\mu}g/g$</TEX>으로 예측하였다. 최적 수율로 예측된 결과는 각각의 조건에 따른 실험을 통해 그 예측의 정확도를 확인하였으며 <TEX>$17.77{\mu}g/g$</TEX>으로 예측조건과 비슷한 결과를 보였다. A 17-run Box-Behnken design (BBD) was used to optimize the extraction conditions of astaxanthin from shrimp waste. Three factors such as ratio of ethanol to raw material, extraction temperature (<TEX>$^{\circ}C$</TEX>) and extraction time (min) were investigated. The adjusted coefficient of determination (<TEX>$R^2{_{adj}}$</TEX>) for the model was 0.9218, and the probability value (p=0.0003) demonstrated a high significance for the regression model. The optimum extraction conditions were found to be: optimized ratio of ethanol to raw material 29.7, extraction temperature <TEX>$49.5^{\circ}C$</TEX> and extraction time 59.9 min. Under these conditions, the mean extraction yield of astaxanthin was <TEX>$17.80{\mu}g/g$</TEX>, which was in good agreement with the predicted model value. Under these conditions, validation experiments were done and the mean extraction yield of astaxanthin was <TEX>$17.77{\mu}g/g$</TEX>, which is in good agreement with the predicted model value.

The ultrasound-assisted enzymatic extraction (UAEE) in this work was utilized to extract triterpenoid from Ganoderma lucidum (G. lucidum). The enzyme concentration (%), ethanol:material ratio (mL/g), ultrasonic duration (min), ultrasonic temperature (°C), and ultrasonic power (W) were all studied as factors impacting triterpenoid content. The experiment was developed using the Plackett–Burman model to filter out the influencing factors and choose the component that has the greatest impact on the triterpenoid content in order to improve the extraction conditions using the Box–Benhken model. The result showed that the highest triterpenoid content of 11.65 mg/g could be obtained under extraction conditions including enzyme concentration of 2.5 %, ethanol:material ratio of 20:1 mL/g, and ultrasonic power of 480 W. The results of gas chromatography-mass spectrometry and Fourier transform infrared spectrometry analysis showed the presence of more phytochemicals in the extract obtained using UAEE compared with the maceration method. Scanning electron microscopy images were used to confirm the assistance of enzymes and ultrasound waves to disrupt the cell walls of G. lucidum. The antibacterial property of the extract by UAEE was evaluated against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella enterica and able to suppress the development of hepatocellular carcinoma, breast carcinoma, and epidermic carcinoma.

The study aimed to develop a new reverse-phase high-performance liquid chromatography (RP-HPLC) method with diode array detection (DAD) detection for simultaneous estimation of escitalopram (EST) and clonazepam (CZP) in tablet dosage forms with a quality by design (QbD) approach. The chromatographic conditions were optimized by Box-Behnken design (BBD) and developed method was validated for the linearity, system suitability, accuracy, precision, robustness, sensitivity, and solution stability according to International Council for Harmonization (ICH) guidelines. EST and CZP standard drugs peaks were separated at retention times of 2.668 and 5.046 min by C-18 column with dimension of 4.6 × 100 mm length and particle size packing 2.5 µm. The mobile phase was methanol: 0.1% orthophosphoric acid (OPA) (25:75, v/v), with a flow rate of 0.7 mL/min at temperature of 26 °C. The sample volume injected was 20 µL and peaks were detected at 239 nm. Using the standard calibration curve, the % assay of marketed tablet was founded 98.89 and 98.76 for EST and CZP, respectively. The proposed RP-HPLC method was able to detect EST and CZP in the presence of their degradation products, indicating the stability-indicating property of the developed RP-HPLC method. The validation parameter’s results in terms of linearity, system suitability, accuracy, precision, robustness, sensitivity, and solution stability were in an acceptable range as per the ICH guidelines. The newly developed RP-HPLC method with QbD application is simple, accurate, time-saving, and economic.

The purpose of this study was to increase the yield and purity of white tea polysaccharides (WTPS) by using traditional techniques available for the separation. Response surface methodology was implemented to predict the optimum conditions for the extraction of polysaccharides in white tea. A Box–Behnken design was used to monitor the effect of extraction temperatures (55 °C, 60 °C, 65 °C, X1), ratio of water solutions to material (10, 12, 14 mL/g, X2), duration of extraction time (60, 90, 120 min, X3) and number of extractions (2, 3, 4, X4) on the dependent variable yield of WTPS (Y, %). In extraction under pre-established conditions, the correlation coefficients (R2) and Adj-R2 of the models for the extraction of WTPS were 0.9932 (P < 0.001) and 0.9872, respectively. As a result, the optimum conditions for WTPS extraction were the following: ratio of water solutions to raw material – 12.48 mL/g, extraction temperature – 54.1 °C, number of extractions – 4 and duration of extraction time – 97.8 min.

The objective of the present study is to find out the optimum extraction conditions for extraction of polyphenols from red grapes using Box–Behnken design. Red grapes polyphenols were extracted using acid–ethanol solvent at various extraction temperature (40–60°C), extraction time (20–100 min) and different solid–liquid ratio (1:5–1:15 g:ml). The effect (main and interactive) of extraction conditions on total anthocyanin, phenolic and flavonoid content were studied using Box–Behnken design (three factors at three levels). The results showed that the contribution of the quadratic model was significant for all the responses. Second-order mathematical regression models were developed and were found to fit well with observed data. Derringer's desirability function methodology was performed to find out the optimal conditions based on both individual and combinations of all responses (extraction temperature: 57°C, time: 61 min, and solid–liquid ratio: 1:8.7 g:ml) were established. At this optimal condition, the anthocyanin yield, total phenolic and flavonoid content were 73.92 mg/100 g, 221.4 mg GAE/100 g, and 79.08 mg CE/100 g, respectively. A desirability value of 0.902 was achieved at this point.

Optimization of ultrasonic-assisted extraction (UAE) of phenolics and antioxidant compounds from rhizomes of Rheum moorcroftianum using response surface methodology (RSM)

Optimum extraction conditions for extraction of soluble rice bran fiber (SRBF) from defatted rice bran (DRB) were determined using response surface methodology (RSM). The three independent variables for extracting SRBF, including ratio of Ca(OH)2 solution to DRB, concentration of Ca(OH)2 and extraction temperature on the yield of SRBF were determined on the basis of single factor experiments. A Box–Behnken design (BBD) was used to investigate the effects of the three independent variables, and the BBD consisted of 15 experimental points with three replications at the center point. The experimental study examined the effect of the independent variables within the following ranges: Ca(OH)2 solution to DRB ratio range of 25:1–30:1, Ca(OH)2 concentration in the range of 2–3% and the temperature in the range of 20–100C The optimum experimental yield of SRBF (7.86 ± 0.04%) was obtained at the extraction condition of 3% Ca(OH)2 solution with a ratio of solution to DRB of 29.75:1 (mL/g) stirred for 1 h at 84C. The experimental yield percentage was in agreement with predicted yield percentage of SRBF by RSM at the optimum extraction conditions. Practical Applications The optimum soluble rice bran fiber (SRBF) yield cannot be estimated based on just the one-factor-at-a-time (single-factor) approach, because it is not only time consuming, but also the interaction between extractions condition may not be included in the determination of the yield and/or quality. A study is needed not only to determine the optimum extraction conditions to obtain a desirable yield or quality of SRBF from defatted rice bran (DRB) but also to understand the degree of interactions between extraction conditions such as temperature, time, ratio of solvent to raw material and solvent concentration.

An ultrasound-assisted procedure for the extraction of water-soluble polysaccharides from Enteromorpha tubulosa was studied by using response surface methodology. Three independent variables including extraction time, extraction temperature, and ratio of liquid to raw material were investigated by single-factor experiments. A three-factor-three-level Box–Behnken design was adopted to optimize the extraction process parameters. A highly significant second-order polynomial equation was fitted by using multiple regression analysis. The optimal extraction conditions were obtained as follows: extraction time 36 min; extraction temperature 69 °C; and ratio of liquid to raw material 45 mL/g. By experiments with these conditions, the extraction yield was 16.04%, which was well matched with the predicted value. The obtained polysaccharides were evaluated for cytotoxicity in vitro against human breast cancer cell line MCF-7 and the results showed that the polysaccharides possessed potent cytotoxicity. Practical applications Seaweeds are one of the largest numbers and species of marine plants, and their bioactive compounds, especially algal polysaccharides, have become the focus of research in the field of food and medicine. Moreover, many algal polysaccharides have been gradually applied to food and medicine owing to their varied biological activities, such as antivirus, antitumor, antiradiation, antimutation, antioxidation, and enhancing immunity. However, there is no universal extraction process for polysaccharides from the different seaweeds because of the different structures of the seaweeds and the varying compositions of algal polysaccharides. In the current research, an ultrasound-assisted procedure for the extraction of water-soluble polysaccharides from Enteromorpha tubulosa was studied by using response surface methodology, and the optimal extraction conditions were obtained so as to provide references for the mass productions in the factories.