- Email: [email protected]
Antioxidant phenolic compounds from Pinus morrisconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters
ARTICLE IN PRESS
JID: JTICE
[m5G;December 5, 2016;15:52]
Journal of the Taiwan Institute of Chemical Engineers 0 0 0 (2016) 1–8
Contents lists available at ScienceDirect
Journal of the Taiwan Institute of Chemical Engineers journal homepage: www.elsevier.com/locate/jtice
Short communication
Antioxidant phenolic compounds from Pinus morrisconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters Pai-Shih Chiang a, Duu-Jong Lee a,b,∗, Chris. G. Whiteley c, Chun-Yung Huang d a
Department of Chemical Engineering, National Taiwan University, Taipei 106, Taiwan Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 106, Taiwan Graduate Institute of Applied Science and Technology, National Taiwan University of Science and Technology, Taipei 106, Taiwan d Department of Seafood Science, National Kaohsiung Marine University, No. 142, Hai-Chuan Rd., Nan-Tzu District, Kaohsiung 811, Taiwan b c
a r t i c l e
i n f o
Article history: Received 11 July 2016 Revised 1 October 2016 Accepted 6 October 2016 Available online xxx Keywords: Pinus morrisonicola Phenolic compounds extraction Antioxidant activity Compressional-puffing pretreatment Catechin Response surface methodology
a b s t r a c t This study is aimed to increase the extraction yield and efficiency of the antioxidant phenolic compounds from Pinus morrisonicola. The optimum extraction conditions for the phenolic compounds from pine needles are determined by using response surface methodology with a central-composite design. Effects of ethanol concentration, extraction temperature and liquid–solid ratio on the extraction yield (total flavonoids content and total phenolic content) and antioxidant activity (2,2 -diphenyl-1picryhydrazyl (DPPH) scavenging activity, ferric reducing antioxidant power (FRAP) and 2,2 -Azino-bis(3ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) scavenging activity) are investigated. The optimum conditions to maximize the extraction yields and antioxidant activity of the pine needles extracts are: 36.1% v/v of ethanol, 70 ºC and 50 ml/g of liquid–solid ratio. The pine needles extracts are shown to be free of 311 tested pesticide residues or certain heavy metals and exhibit comparable quantities of total flavonoids contents, total phenolic contents and the antioxidant activities with commercial products. © 2016 Taiwan Institute of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
1. Introduction Pinus morrisonicola is an indigenous pine growing at elevations of 30 0–230 0 m in the Taiwanese mountain areas. In ancient Chinese medicine books, the curative effects of various parts of the pine were documented. In particular, extracts of pine needles had been revealed to exhibit antioxidant activity as food supplements [1–3]. Antioxidant activity is defined as the compounds with an ability to delay, inhibit, or prevent the oxidation of materials by scavenging free radicals and diminishing oxidative stress [4–6]. The benefits derived from phenolic compounds, including phenolic acids, flavonoids, tannins and the less common stilbenes and lignans, have been referred to their antioxidant activity, and phenolic compounds could be regarded as the major determinant of antioxidant potentials of foods [6–9]. Solvent extraction is commonly used to extract bioactive compounds from plants [10,11]. Pine needles extracts were proposed to be produced by solvent extraction, mostly using ethanol–water ∗ Corresponding author at: Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 106, Taiwan. Fax: +886 2 23623040. E-mail addresses: [email protected], [email protected] (D.-J. Lee).
mixture as the solvent [12–17]. Extraction starts when solvent molecules penetrate the plant matrices, causing the cytoplasm layer to be exposed directly to the solvent, bringing about the dissolution of the bioactive compounds [18]. The mixture of ethanol and water at different concentrations is used as an extraction solvent in the extraction of bioactive compounds from plants [19–21]. The ethanol concentration affects the dielectric constant of the mixture hence altering the energy required to deteriorate the attraction of water molecules to the target molecules. The extraction temperature is also a critical factor to the stability of bioactive compounds and the extraction performance. High temperature is preferred for extraction of thermally stable compounds from plants since high temperature leads to an increase of the solute diffusivity and decrease of the energy barrier [22]. Conversely, the temperature of extraction should be limited for extracting thermal sensitive compounds. Liquid–solid ratio is also an important extraction parameter and when applied correctly can decrease the mass transfer barrier during the diffusion of active compounds and subsequently enhance the extraction yield [23]. However, if the ratio is beyond the optimum, the excess solvent does not have a significant effect on the equilibrium extraction yields and would lead to a solvent waste.
http://dx.doi.org/10.1016/j.jtice.2016.10.010 1876-1070/© 2016 Taiwan Institute of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010
ARTICLE IN PRESS
JID: JTICE 2
[m5G;December 5, 2016;15:52]
P.-S. Chiang et al. / Journal of the Taiwan Institute of Chemical Engineers 000 (2016) 1–8
Compressional puffing is a fast and inexpensive method for drying fruits and vegetables [24], and the process is carried out in a continuous puffing system at elevated pressure and in a superheated steam, which rapidly brings the water within the samples to a temperature above its atmospheric boiling point. When the samples are immediately exposed to atmospheric pressure, the intratissue vapor flashes to form a porous structure of plant biomass [25]. Huang et al. [26] introduced the process for bioactive compounds extraction due to the decomposition of the cellular structure of plant during the process. Also, comparing the other pretreatments such as ion-liquid, ultrasonic and microwave-assisted pretreatment, compressional-puffing provides advantages including relative simple procedure, reactant-saving, reduced pollution and feasibility for continuous production, while having great enhancement in extraction yield. P. morrisonicola products are commercially available which are based on time-consuming, ill-controlled biological hydrolysis reactions. The present study is the first report to note the optimal extraction parameter sets for hydrolyzed pine needles using compressional-puffing pretreatment. Since the puffing process shortens the treatment time from years to 10 s, the tested process is highly promising for yielding cost-effective and well-controlled digestate product by pine needle. The optimum extraction conditions for the phenolic compounds from pine needles were determined by using response surface methodology with a centralcomposite design. The effects of ethanol concentration, extraction temperature and liquid–solid ratio on the extraction yield (total flavonoids content and total phenolic content) and antioxidant activity (2,2 -diphenyl-1-picryhydrazyl (DPPH) scavenging activity, ferric reducing antioxidant power (FRAP) and 2,2 -Azino-bis(3ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) scavenging activity) are investigated. A quadratic model afforded a superior fit of the experimental data with r2 > 0.98 and p value <0.01. The optimum conditions to maximize the extraction yields and antioxidant activity of the pine needles extracts are: 36.1% v/v of ethanol, 70 °C and 50 ml/g of liquid–solid ratio. In addition, the pine needles extracts are also compared with commercial enzyme products and show competitiveness in both total phenolic content and antioxidant activity. 2. Materials and methods 2.1. Materials
with puffing temperature 220 °C. The unit possessed three compression (to 5 kg/cm2 )–relaxation cycles with cycle time of 10 s. After the three cycles, the chamber was opened to induce puffing.
2.3. Solvent extraction tests 2.3.1. Experimental design The treated pine needles were extracted in different extraction environments involving three variables (extraction temperature, ethanol concentration and liquid–solid ratio). A three-factor central composite design (CCD) was used to identify the relationship existing between the response function and the extraction process variables, as well as to determine those conditions that optimized the extraction process of total extraction content and the antioxidant activity of the pine needle extracts. The independent variables studied in this experiment were ethanol concentration (X1 : 25–75%), extraction temperature (X2 : 50–80 °C), and liquid–solid ratio (X3 : 23–57 ml/g), while response variables were total flavonoid content (TFC) (Y1 ), total phenol content (TPC) (Y2 ), DPPH radical scavenging activity (Y3 ), ferric reducing antioxidant power (Y4 ) and ABTS radical scavenging activity (Y5 ). The selection and range of these three factors was based on the premier experiment data. Each factor was coded at three levels (−1, 0 and 1) (Table S1). Twenty randomized experiments included six replicates as the center points were assigned based on CCD (Table 1).
2.3.2. Statistical analysis The Design Expert Version 10.0 (Stat-Ease, Minneapolis, MN, USA) software was used to conduct the experimental design and the statistical analysis. Results for the total content and antioxidant activity were expressed as means ± standard deviations. A response surface analysis and analysis of variance (ANOVA) were employed to determine the regression coefficients, statistical significance of the model terms and to fit the mathematical models of the experimental data that aimed to optimize the overall region for both response variables. The model proposed for the response is given below:
Yi = b0 + b1 X1 + b2 X2 + b3 X3 + b11 X12 + b22 X22 + b33 X32 + b12 X1 X2 + b13 X1 X3 + b23 X2 X3
(1)
The pine needles were collected from Nantou County, Taiwan. These needles were washed with tap water and then dried at room temperature to 6% w/w moisture content. The liquid samples from two commercial “enzyme products”, namely, Yamato Enzyme from Yamato Kouso Co., Ltd, Osaka, Japan and Gold Enzyme from Golden Enzyme, Nantou County, Taiwan, were used as the samples for comparison. Chemicals 6-Hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox), 2,2 -Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 2,2 -diphenyl-1-picryhydrazyl (DPPH), 2,4,6-Tripyridyl-s-Triazine (TPTZ), ferric chloride, formic acid (95%), gallic acid and iron(II) sulfate heptahydrate were from SigmaAldrich (St. Louis, USA) and acetonitrile was from Burdick and Jackson (New Jersey, USA). The Folin–Ciocalteu reagent was from Merck (Darmstadt, Germany). Quercetin was from Acros Organics (New Jersey, USA).
where Yi is predicted response, b0 is offset term that fixes the response at the central point; b1 , b2 and b3 are linear effect terms, b11 , b22 and b33 are the quadratic effect terms; and b12 , b23 and b13 are interaction effect terms. The proportion of variance explained by the polynomial models obtained is given by the multiple coefficients of determination, r2 , and the ANOVA analysis. The significance of each coefficient was determined using the Students t-test and p value, and nonsignificant coefficients were removed to obtain a reduced model. The optimum condition was verified by conducting experiments under these conditions. Responses were monitored and results compared with model predictions. The fitted polynomial equation was expressed as surface plots in order to visualize the relationship between the response and experimental levels of each factor and to deduce the optimum conditions.
2.2. The compressional-puffing treatment
2.3.3. Verification of the model Experimental data for the total content and antioxidant activity were obtained according to the recommended optimum conditions. The responses were determined after extraction of phenolic compounds under optimal conditions. The experimental and
The pine needle samples (<5 mm) were placed in compressional-puffing chamber MIBO R2 (Yuan Chuang Food Machinery Co. Ltd., Taiwan) [26]. And the pine needs were treated
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010
ARTICLE IN PRESS
JID: JTICE
[m5G;December 5, 2016;15:52]
P.-S. Chiang et al. / Journal of the Taiwan Institute of Chemical Engineers 000 (2016) 1–8
3
Table 1 Three-factor central composite design used for RSM with experimental and predicted values for the independent variables. Run
Coded factors X1
X2
Response X3
Y1 (TFC) exp
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 Run
−1.00 1.00 −1.00 −1.00 −1.00 1.00 1.00 1.00 0.00 0.00 0.00 0.00 −1.68 1.68 0.00 0.00 0.00 0.00 0.00 0.00
−1.00 −1.00 1.00 −1.00 1.00 −1.00 1.00 1.00 0.00 0.00 0.00 0.00 0.00 0.00 −1.68 1.68 0.00 0.00 0.00 0.00
Predicted
26.68 25.94 36.34 28.95 37.54 27.99 33.02 32.23 29.91 32.77 29.32 30.44 23.93 19.83 29.27 45.32 8.15 55.88 31.12 29.46
26.04 25.20 36.98 29.02 36.36 28.18 32.66 32.04 30.48 30.48 30.48 30.48 24.34 20.25 29.66 45.69 8.56 56.24 30.48 30.48
exp 29.79 28.17 39.70 33.28 43.09 31.50 32.90 35.54 32.50 31.83 29.91 30.40 24.64 18.81 28.65 45.39 8.34 61.56 32.50 31.10
Predicted 28.71 27.03 38.57 31.93 41.79 30.25 31.41 34.63 31.29 31.29 31.29 31.29 26.35 20.54 30.35 47.08 10.07 63.23 31.29 31.29
Response Y3 (DPPH)
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
−1.00 −1.00 −1.00 1.00 1.00 1.00 −1.00 1.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 −1.68 1.68 0.00 0.00
Y2 (TPC)
Y4 (FRAP)
Y5 (ABTS)
exp
Predicted
exp
Predicted
exp
96.6 85.5 112.5 123.0 139.5 95.6 103.4 98.1 105.6 118.1 113.2 117.7 99.0 59.8 111.9 152.2 30.5 206.1 121.9 115.9
95.4 83.5 132.7 102.5 139.7 90.6 90.4 97.5 103.3 103.3 103.3 103.3 100.8 55.3 112.1 149.1 15.6 191.1 103.3 103.3
241.2 227.1 243.7 267.6 283.5 243.1 251.3 247.2 240.3 253.8 250.1 251.4 246.0 188.0 247.9 365.5 87.8 387.4 259.7 249.8
242.1 230.5 245.4 266.8 286.9 244.9 250.6 248.1 251.0 251.0 251.0 251.0 244.2 186.2 246.0 363.4 86.1 385.4 251.0 251.0
4563 4263 6462 5844 7496 3781 5535 6020 4978 5103 5117 5089 4052 3188 5713 8194 1079 9742 5301 5213
predicted values were compared in order to determine the validity of the model.
2.4. Chemical analyses 2.4.1. TFC and TPC measurements The TFC was determined using the method of Park et al. [27] with modification. 0.5 ml of the extract solution was added to mixture of 5 ml ethanol, 0.25 ml 0.5 M sodium nitrite solution and 0.25 ml of 0.3 M aluminum chloride. Then, 1.5 ml of 1 M sodium hydroxide was added to the mixture. The absorbance of the mixture was measure at 510 nm with an UV–vis spectrophotometer (DR2700TM portable spectrophotometer, HACH, Colorado, USA) using quercetin as indicator. The TPC was measured using the method of Adefegha [28] with modification. 0.1 ml of the extract solution was added to mixture of 0.5 ml of Folin–Ciocalteu’s reagent, 5.9 ml deionized water. Then, 1.5 ml of 20% w/w sodium carbonate was added. The mixture was kept at 40 °C for 30 min. Then the absorbance of the mixture was
Predicted 4506 4196 6395 5787 7429 3714 5478 5963 5128 5128 5128 5128 4142 3279 5798 8277 1171 9825 5128 5128
measure at 765 nm with an UV–vis spectrophotometer using gallic acid as indicator. 2.4.2. Antioxidant activity assay The DPPH method was conducted for extract samples using the method proposed by Brand-Williams et al. [29]. 0.1 ml of example was added to 3.9 ml of the DPPH solution with vortexed for 1 min and left in the dark for 30 min at room temperature. The absorbance of samples was measured at 517 nm using an UV–vis spectrophotometer using Trolox as indicator. The FRAP potential of the samples was determined by the method of Benzie and Strain [30]. 1.5 ml FRAP solution (250 mM acetate buffer, 0.83 mM TPTZ (2,4,6-tripyridyl-s-traizine), 3.33 mM hydrochloride acid and 1.67 mM ferric chloride) was warmed to 37 °C and was mixed with 50 μl of the sample and 150 μl of the deionized water. The sample was incubated at 37 °C for 4 min and the absorbance at 593 nm using UV–vis spectrophotometer was recorded using ferrous sulfate as indicator. The ABTS radical scavenging activity for extract samples was determined based on the method of Arnao et al. [31]. 150 μl of the
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010
ARTICLE IN PRESS
JID: JTICE 4
[m5G;December 5, 2016;15:52]
P.-S. Chiang et al. / Journal of the Taiwan Institute of Chemical Engineers 000 (2016) 1–8 Table 2 Quadratic equations and statistical parameters calculated after implementation of central composite experimental design. Response
2nd order polynomial equation
R2
R2 (adjusted)
P value
Lack of fit
Y1 (TFC)
30.48 − 1.22X1 + 4.77X2 14.19X3 − 0.87X1 X2 −0.49X2 X3 − 2.9X1 X1 + 2.55X2 X2 + 0.68X3 X3 31.29 − 1.73X1 + 4.9X2 + 15.82X3 + 1.37X1 X2 −2.78X1 X1 + 2.32X2 X2 + 1.9X3 X3 103.34 − 13.54X1 + 11.03X2 + 52.21X3 + 2.25X1 X2 −9.84X1 X3 − 8.97X1 X1 + 9.65X2 X2 250.97 − 17.24X1 + 34.94X2 + 89.06X3 −6.79X1 X3 − 12.68X1 X1 + 19.04X2 X2 − 5.4X3 X3 5128.47 − 256.87X1 + 737.61X2 + 2575.53X3 −289X1 X3 + 90X2 X3 − 502.25X1 X1 + 676.32X2 X2 + 130.79X3 X3
0.9938
0.9833
<0.01
0.1963
0.9839
0.9596
<0.01
0.1223
0.9713
0.9312
0.0248
0.0496
0.9964
0.9885
<0.01
0.3903
0.9978
0.9930
<0.01
0.1542
Y2 (TPC) Y3 (DPPH) Y4 (FRAP) Y5 (ABTS)
extracts were reacted with 2850 μl of the ABTS solution at dark for 2 h. Then the absorbance was taken at 734 nm using the spectrophotometer using Trolox as indicator. All determinations were estimated in triplicate. 2.4.3. Other analyses Chemical composition of the phenolic compounds extracted from P. morrisonicola was analyzed by liquid chromatography–mass spectrometry (LC–MS) method. Chromatographic separation was performed by using the following binary gradient of two mobile phases, (A): 0.1% (v/v) aqueous formic acid and (B): acetonitrile consisting of 0.1% (v/v) formic acid, at a flow rate of 300 μl/min: 10% B (0 min), 10% B (10 min), 15% (20 min), 40% B (35 min). The UV detector was set at the wavelength of 280 nm with column temperature at 25 °C and the injection volume at 20 μl for all samples. The identifications of the active functional groups presence in the extracts from P. morrisonicola is conducted by FTIR spectroscopy (Perkin Elmer Spectrum 20 0 0, USA) over the wavelength range of 40 0 0–60 0 cm−1 . 3. Results and discussion 3.1. Extract characteristics Extract from P. morrisonicola, HPLC chromatograms and FTIR spectrums were conducted in order to characterize the components of extracted phenolic compounds. The HPLC chromatogram of pine needles extract and that of pure (+)-catechin hydrate were presented in Fig. S1, showing that both exhibited as major peak at 12 min. The FTIR spectra for the pine needle extract and for (+)-catechin hydrate were shown in Fig. S2. Based on these analyses, the present pine needle extract contain catechinlike substances enriched with O–H bond (3339 cm−1 ), C=O bond (1694 cm−1 ), aromatic ring (1603, 1516 and 1451 cm−1 ), and C–O–C bond (1257 cm−1 ). The pine needle extract samples were sent to SGS Taiwan Co. Ltd for characterizing quantities of 311 pesticide residues and that of copper (Cu), arsenic (As), lead (Pb), mercury (Hg) or cadmium (Cd). The results revealed that the extracts were free of these pesticide residues or heavy metals (Supplementary materials) and is regarded safe for human consumption. 3.2. Optimization of extract yields and antioxidant activities 3.2.1. Model fitting Experimental values of extraction yield (TFC and TPC) and antioxidant activity (DPPH scavenging activity, FRAP and ABTS scavenging activity) were used in a multiple regression analysis using response surface methodology to fit the quadratic equations (Eq. (1)). In this study, all experimental data were obtained from a 20-run-experiment with the pine needles pretreated with 220 °C
compressional-puffing, and predicted data were from a response surface analysis model with the results shown in Table 1. An ANOVA analysis for the quadratic model was used to test the significance and accuracy of the model. Highly significant levels for these (p < 0.01) were obtained by statistical analysis, while insignificant variables were ignored in order to get a precise and applicable model. The model, established by the regression equations, replaces the experimental real point to explain the response results. The regression equations, coefficients of determination (r2 ), adjusted r2 values, probability values (p) and lack of fit values for dependent variables are shown in Table 2. The results suggested that the extraction yield and antioxidant activity were primarily determined by the linear and quadratic terms. In addition, the predicted results according to the models were close to the observed experimental responses, indicating that the generated models adequately explained the data variation and can represent the actual relationships between the selected variables. 3.2.2. Effect of extraction parameters on extraction yield The data shown in Table 1 indicate that the extraction yield (total flavonoids content and total phenolic content) of the pine needles extracts and the extraction parameters were quadratic with a good coefficient (r2 = 0.9833 and 0.9596, respectively). The effect of ethanol concentration and extraction temperature on the total flavonoids content (TFC) is shown (Fig. 1a). The extraction yield increases with extraction temperature with a fixed ethanol concentration. High extraction temperature could lead to an increase in yield of the phenolic compounds through increasing phenolic solubility, decreased solvent viscosity and surface tension. Also the experiment was conducted between 50 and 70 °C since at higher temperatures there is degradation and ionization of anthocyanin which leads to a decrease in extraction yield and antioxidant activity [32]. The linear trend of the model with respect to extraction temperature is expected (Fig. 1c) The effect of ethanol concentration on extraction yield shows a quadratic fitting as a high peak is realized in the response surface of the effect of ethanol concentration and extraction temperature (Fig. 1a) and the effect of ethanol concentration and liquid–solid ratio (Fig. 1b). It is also in good agreement with previous section in other studies [33]. The effect of liquid–solid ratio is shown according to the response surface plots (Fig. 1b and c). Due to the fact that more solvent can enter plant cells while more flavonoids can permeate into the solvent under the higher liquid–solid ratio, a high ratio leads to greater extraction yield. The response of total phenolic content (TPC) shown (Fig. 1d–f) indicate a similar tendency and result with response to TFC. In conclusion, in order to maximize the extraction yield from P. morrisconicola, 70 °C extraction temperature, 50 ml/g of liquid–solid ratio and a moderate ethanol concentration should be adopted. 3.2.3. Effects of extraction parameters on antioxidant activities The data shown in Table 2 indicates that antioxidant activities (DPPH scavenging ability, FRAP and ABTS scavenging ability) of the
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010
JID: JTICE
ARTICLE IN PRESS
[m5G;December 5, 2016;15:52]
P.-S. Chiang et al. / Journal of the Taiwan Institute of Chemical Engineers 000 (2016) 1–8
5
Fig. 1. Response surface plot for the parameters considered. (a) Temperature and ethanol concentration on TFC; (b) liquid–solid ratio and ethanol concentration on TFC; (c) liquid–solid ratio and temperature on TFC; (d) temperature and ethanol concentration on TPC; (e) liquid–solid ratio and ethanol concentration on TPC; (f) liquid–solid ratio and temperature on TPC.
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010
JID: JTICE 6
ARTICLE IN PRESS
[m5G;December 5, 2016;15:52]
P.-S. Chiang et al. / Journal of the Taiwan Institute of Chemical Engineers 000 (2016) 1–8
Fig. 2. Response surface plot for the parameters considered. (a) Temperature and ethanol concentration on DPPH scavenging activity; (b) liquid–solid ratio and ethanol concentration on DPPH scavenging activity; (c) liquid–solid ratio and temperature on DPPH scavenging activity; (d) temperature and ethanol concentration on FRAP; (e) liquid–solid ratio and ethanol concentration on FRAP; (f) liquid–solid ratio and temperature on FRAP; (g) temperature and ethanol concentration on ABTS scavenging activity; (h) liquid–solid ratio and ethanol concentration on ABTS scavenging activity; (i) liquid–solid ratio and temperature on ABTS scavenging activity.
pine needles extracts and the extraction parameters are quadratic. The response surface plots are shown (Fig. 2a and i), and they show a great similarity with the results of extraction yield. Antioxidant activity of the pine needles extract appears to have the highest response in the range of 40–50% v/v ethanol concentration with respect to different extraction parameters. Extraction temperature causes a linear increase in the antioxidant activity which is similar to the result of extraction yield. Liyana-Pathirana and Shahidi [34] reported that the extraction rate of thermally stable antioxidants at elevated temperature was higher than the decomposition of less soluble antioxidants, which could be confirmed in the present study. The chemical degradation and decomposition during the extraction is the main mechanism leading to the decrease in extraction yield as well as antioxidant activity [35], therefore, a proper setting of extraction temperature could result in a maximum response, reported as 70 °C in this study. Liquid-to-solid ratio effects on the antioxidant activity of the pine needles extracts are much more significant than other extraction parameters due to the result of extraction yield [36] (see Table 3). 3.2.4. Verification of the models and optimization of extraction yield and antioxidant activity Table 1 lists that the experimental values were close to the predicted values. The optimal conditions obtained using the models
were as follows: 36.1% v/v ethanol concentration, 70 °C extraction temperature and 50 ml/g liquid–solid ratio. Under optimal conditions, the model predicted a maximum response of 39.6 quercetin equivalent (QE) mg/g DW of TFC, 44.1 gallic acid equivalent (GAE) mg/g DW of TPC, 151.8 Trolox equivalent (TE) μmol/g DW of DPPH scavenging activity, 315.5 mmol FeSO4 /g DW of FRAP and 7464 TEAC mg/g DW of ABTS scavenging activity. The extraction conducted at the optimal conditions yield: 38.4 ± 1.3 QE mg/g DW of TFC, 46.9 ± 0.7 GAE mg/g DW of TPC, 167.2 ± 3.3 TE μmol/g DW of DPPH scavenging activity, 309.9 ± 7.8 mmol FeSO4 /g DW of FRAP and 7277 ± 102 TEAC mg/g DW of ABTS scavenging activity. The experimental results correlate well with the model prediction. 3.2.5. Comparison with commercial products The extract from pine needs at the optimal extraction conditions (36.1% v/v aqueous ethanol, 70 °C and 50 ml/g of liquid–solid ratio) from 220 °C-puffing pretreated pine needles was compared its TPC, TFC and DPPH scavenging activity with those of the extract from original pine needles and from the two commercial products. At optimal extraction conditions, the puffing pretreatment enhanced TFC from 0.154 to 0.768 QE mg/ml, TPC from 0.211 to 0.938 GAE mg/ml, and DPPH scavenging activity from 1264 to 3344 TE μM. The Yamato Enzyme and Golden Enzyme contain no TFC but have 0.716 and 0.899 GAW mg/ml of TPC, respectively. The
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010
ARTICLE IN PRESS
JID: JTICE
[m5G;December 5, 2016;15:52]
P.-S. Chiang et al. / Journal of the Taiwan Institute of Chemical Engineers 000 (2016) 1–8
7
Table 3 Comparison of pine needles extracts with commercial products on phenolic compounds contents and antioxidant activity.
Untreated pine needles 220 °C-puffing pretreated pine needles Yamamoto enzyme (from fruit and vegetables with 60-d processing time) Golden enzyme (from fruit and vegetables with 90-d processing time)
corresponding DPPH scavenging activity data are 3198 and 2763 TE μM, respectively. Restated, the present pretreated-extracted sample has comparable quantity of TPC and antioxidant activity as the two commercial products. It is worth noting that the two tested commercial products were made from fruit and vegetables at a few months processing time. Conversely, the present sample is from pine needles, a biomass that are not consumable by human beings, unlike the fruit and vegetables adopted by the commercial products. Additionally, the total processing time for the present samples is within 24 h, much shorter than the long fermentation times needed for the commercial enzyme products. The tested process can yield extract with high TFC/TPC quantities of high antioxidant activities with short processing time.
4. Conclusions The optimal extraction parameters for puffing-pretreated P. morrisonicola to yield maximum PTC and PFC equivalents and autioxidant activities were identified using response surface methodology. Under optimum conditions (70 °C extraction temperature, 50 ml/g liquid–solid ratio and 36.1% v/v ethanol concentration), the extract has the following readings: 38.4 ± 1.3 QE mg/g DW of TFC, 46.9 ± 0.7 GAE mg/g DW of TPC, 167.2 ± 3.3 TE μmol/g DW of DPPH scavenging activity, 309.9 ± 7.8 mmol FeSO4 /g DW of FRAP and 7277 ± 102 TEAC mg/g DW of ABTS scavenging activity. The present extract from pine needles was noted to have comparable TPC equivalent and DPPH scavenging activities as noted from commercial products derived from fruit and vegetables.
Supplementary materials Supplementary material associated with this article can be found, in the online version, at doi:10.1016/j.jtice.2016.10.010. References [1] Kong ZM, Liu ZT, Ding BY. Study on the antimutagenic effect of pine needle extract. Mutation Res Lett 1995;347:101–4. [2] Yen GC, Duh PD, Huang DW, Hsu CL, Fu TYC. Protective effect of pine (Pinus morrisonicola Hay.) needle on LDL oxidation and its anti-inflammatory action by modulation of iNOS and COX-2 expression in LPS-stimulated RAW 264.7 macrophages. Food Chem Toxicol 2008;46:175–85. ˇ [3] Schäfer A, Chovanová Z, Muchová J, Sumegová K, Liptáková A, Dura cˇ ková Z, et al. Inhibition of COX-1 and COX-2 activity by plasma of human volunteers after ingestion of French maritime pine bark extract (Pycnogenol). Biomed Pharm 2006;60:5–9. [4] Yan JK, Wang YY, Ma HL, Wang ZB, Pei JJ. Structural characteristics and antioxidant activity in vivo of a polysaccharide isolated from Phellinus lintenus mycelia. J Taiwan Inst Chem Eng 2016;65:110–17. [5] Li K, Yang X, Hu X, Han C, Lei Z, Zhang Z. In vitro antioxidant, immunomodulatory and anticancer activities of two fractions of aqueous extract from Helicteres angustifolia L. root. J Taiwan Inst Chem Eng 2016;61:75–82. [6] Paun G, Neagu E, Albu C, Moroeanu V, Radu G. Antioxidant activity and inhibitory effect of polyphenolic-rich extract from Betonica officinalis and Impatiens noli-tangera berbs on key enzyme linked to type 2 diabetes. J Taiwan Inst Chem Eng 2016;60:1–7. [7] Heim KE, Tagliaferro AR, Bobilya DJ. Flavonoid antioxidants: chemistry, metabolism and structure-activity relationships. J Nutr Biochem 2002;13:572–84.
TFC (QE mg/ml)
TPC (GAE mg/ml)
DPPH scavenging activity (TE μM)
0.154 ± 0.009 0.768 ± 0.031 NA NA
0.211 ± 0.028 0.938 ± 0.014 0.716 ± 0.042 0.889 ± 0.013
1264 ± 74 3344 ± 165 3198 ± 312 2763 ± 103
[8] Parr AJ, Bolwell GP. Phenols in the plant and in man. The potential for possible nutritional enhancement of the diet by modifying the phenols content or profile. J Sci Food Agric 20 0 0;80:985–1012. [9] Hollman PCH, Katan MB. Dietary flavonoids: intake, health effects and bioavailability. Food Chem Toxicol 1999;37:937–42. [10] Sodeifian G, Sajadian SA, Ardestani NS. Evaluation of the response surface and hybrid artificial neutral network-genetic algorithm methodologies to determine extraction yield of Ferulago angulata through supercritical fluid. J Taiwan Inst Chem Eng 2016;60:166–73. [11] Zhu XW, Wei MP, Xu DP, Guo YH, Xie YF, Yao WR, et al. In vitro and in vivo antitumor effects of the extract of Sapindus spp. J Taiwan Inst Chem Eng 2016;66:27–32. [12] Touriño S, Selga A, Jiménez A, Juliá L, Lozano C, Lizárraga D, et al. Procyanidin fractions from pine (Pinus pinaster) bark: radical scavenging power in solution, antioxidant activity in emulsion, and antiproliferative effect in melanoma cells. J Agric Food Chem 2005;53:4728–35. [13] Kwak CS, Moon SC, Lee MS. Antioxidant, antimutagenic, and antitumor effects of pine needles (Pinus densiflora). Nutr Cancer 2006;56:162–71. [14] Bito T, Roy S, Sen CK, Packer L. Pine bark extract pycnogenol downregulates IFN-γ -induced adhesion of T cells to human keratinocytes by inhibiting inducible ICAM-1 expression. Free Radical Biol Med 20 0 0;28:219–27. [15] Ka MH, Choi EH, Chun AHS, Lee KG. Antioxidative activity of volatile extracts isolated from Angelica tenuissimae roots, peppermint leaves, pine needles, and sweet flag leaves. J Agric Food Chem 2005;53:4124–9. [16] Park YS, Jeon MH, Hwang HJ, Park MR, Lee SH, Kim SG, et al. Antioxidant activity and analysis of proanthocyanidins from pine (Pinus densiflora) needles. Nutr Res Pract 2011;5:281–7. [17] Zeng WC, Zhang Z, Gao H, Jia LR, He Q. Chemical composition, antioxidant, and antimicrobial activities of essential oil from pine needle (Cedrus deodara). J Food Sci 2012;77:824–9. [18] Rakotondramasy-Rabesiaka L, Havet JL, Porte C, Fauduet H. Solid–liquid extraction of protopine from Fumaria officinalis L. – Analysis determination, kinetic reaction and model building. Sep Purif Technol 2007;54:253–61. [19] Khiari Z, Makris DP, Kefalas P. An investigation on the recovery of antioxidant phenolics from onion solid wastes employing water/ethanol-based solvent systems. Food Bioprocess Technol 2009;2:337–43. [20] Durling N, Catchpole O, Grey J, Webby R, Mitchell K, Foo L, et al. Extraction of phenolics and essential oil from dried sage (Salvia officinalis) using ethanol–water mixtures. Food Chem 2007;101:1417–24. [21] Ofori-Boateng C, Lee KT, Saad B. A biorefinery concept for simultaneous recovery of cellulosic ethanol and phenolic compounds from oil palm fronds: Process optimization. Energy Conver Manag 2014;81:192–200. [22] Cissé M, Bohuon P, Sambe F, Kane C, Sakho M, Dornier M. Aqueous extraction of anthocyanins from Hibiscus sabdariffa: Experimental kinetics and modeling. J Food Eng 2012;109:16–21. [23] Franco D, Sineiro J, Pinelo M, Núñez MJ. Ethanolic extraction of Rosa rubiginosa soluble substances: Oil solubility equilibria and kinetic studies. J Food Eng 2007;79:150–7. [24] Kozempel MF, Sullivan JF, Craig JC, Konstance RP. Explosion puffing of fruits and vegetables. J Food Sci 1989;54:772–3. [25] Sullivan JF, Konstance RP, Aceto NC, Heiland WK, Craig JC. Continuous explosion-puffing of potatos. J Food Sci 1977;42:1462–3. [26] Huang CY, Wu SJ, Yang WN, Kuan AW, Chen CY. Antioxidant activities of crude extracts of fucoidan extracted from Sargassum glaucescens by a compressional-puffing-hydrothermal extraction process. Food Chem 2016;197:1121–9. [27] Park YS, Jung ST, Kang SG, Heo BG, Arancibia-Avila P, Toledo F. Antioxidants and proteins in ethylene-treated kiwifruits. Food Chem 2008;107:640–8. [28] Adefegha SA, Oboh G. Enhancement of total phenolics and antioxidant properties of some tropical green leafy vegetables by steam cooking. J Food Process Preserv 2011;35:615–22. [29] Brand-Williams W, Cuvelier ME, Berset C. Use of a free radical method to evaluate antioxidant activity. LWT Food Sci Technol 1995;28:25–30. [30] Benzie IFFF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of ‘Antioxidant Power’: The FRAP assay. Anal Biochem 1996;239:70–6. [31] Arnao MB, Cano A, Acosta M. The hydrophilic and lipophilic contribution to total antioxidant activity. Food Chem 2001;73:239–44. [32] Wang J, Sun B, Cao Y, Tian Y, Li X. Optimisation of ultrasound-assisted extraction of phenolic compounds from wheat bran. Food Chem 2008;106:804– 810. [33] Yang YC, Li J, Zu YG, Fu YJ, Luo M, Wu N, et al. Optimisation of microwave-assisted enzymatic extraction of corilagin and geraniin from Geranium sibiricum Linne and evaluation of antioxidant activity. Food Chem 20 0 0;122:373– 380.
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010
JID: JTICE 8
ARTICLE IN PRESS
[m5G;December 5, 2016;15:52]
P.-S. Chiang et al. / Journal of the Taiwan Institute of Chemical Engineers 000 (2016) 1–8
[34] Liyan-Pathirana C, Shahidi F. Optimization of extraction of phenolic compounds from wheat using response surface methodology. Food Chem 2005;93:47–56. [35] Pompeu DR, Silva EM, Rogez H. Optimisation of the solvent extraction of phenolic antioxidants from fruits of Euterpe oleracea using response surface methodology. Bioresour Technol 20 09;10 0:6076–82.
[36] Prasad KN, Hassan FA, Yang B, Kong KW, Ramanan RN, Azlan A, et al. Response surface optimisation for the extraction of phenolic compounds and antioxidant capacities of underutilised Mangifera pajang Kosterm peels. Food Chem 2011;128:1121–7.
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010
JID: JTICE
[m5G;December 5, 2016;15:52]
Journal of the Taiwan Institute of Chemical Engineers 0 0 0 (2016) 1–8
Contents lists available at ScienceDirect
Journal of the Taiwan Institute of Chemical Engineers journal homepage: www.elsevier.com/locate/jtice
Short communication
Antioxidant phenolic compounds from Pinus morrisconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters Pai-Shih Chiang a, Duu-Jong Lee a,b,∗, Chris. G. Whiteley c, Chun-Yung Huang d a
Department of Chemical Engineering, National Taiwan University, Taipei 106, Taiwan Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 106, Taiwan Graduate Institute of Applied Science and Technology, National Taiwan University of Science and Technology, Taipei 106, Taiwan d Department of Seafood Science, National Kaohsiung Marine University, No. 142, Hai-Chuan Rd., Nan-Tzu District, Kaohsiung 811, Taiwan b c
a r t i c l e
i n f o
Article history: Received 11 July 2016 Revised 1 October 2016 Accepted 6 October 2016 Available online xxx Keywords: Pinus morrisonicola Phenolic compounds extraction Antioxidant activity Compressional-puffing pretreatment Catechin Response surface methodology
a b s t r a c t This study is aimed to increase the extraction yield and efficiency of the antioxidant phenolic compounds from Pinus morrisonicola. The optimum extraction conditions for the phenolic compounds from pine needles are determined by using response surface methodology with a central-composite design. Effects of ethanol concentration, extraction temperature and liquid–solid ratio on the extraction yield (total flavonoids content and total phenolic content) and antioxidant activity (2,2 -diphenyl-1picryhydrazyl (DPPH) scavenging activity, ferric reducing antioxidant power (FRAP) and 2,2 -Azino-bis(3ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) scavenging activity) are investigated. The optimum conditions to maximize the extraction yields and antioxidant activity of the pine needles extracts are: 36.1% v/v of ethanol, 70 ºC and 50 ml/g of liquid–solid ratio. The pine needles extracts are shown to be free of 311 tested pesticide residues or certain heavy metals and exhibit comparable quantities of total flavonoids contents, total phenolic contents and the antioxidant activities with commercial products. © 2016 Taiwan Institute of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
1. Introduction Pinus morrisonicola is an indigenous pine growing at elevations of 30 0–230 0 m in the Taiwanese mountain areas. In ancient Chinese medicine books, the curative effects of various parts of the pine were documented. In particular, extracts of pine needles had been revealed to exhibit antioxidant activity as food supplements [1–3]. Antioxidant activity is defined as the compounds with an ability to delay, inhibit, or prevent the oxidation of materials by scavenging free radicals and diminishing oxidative stress [4–6]. The benefits derived from phenolic compounds, including phenolic acids, flavonoids, tannins and the less common stilbenes and lignans, have been referred to their antioxidant activity, and phenolic compounds could be regarded as the major determinant of antioxidant potentials of foods [6–9]. Solvent extraction is commonly used to extract bioactive compounds from plants [10,11]. Pine needles extracts were proposed to be produced by solvent extraction, mostly using ethanol–water ∗ Corresponding author at: Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 106, Taiwan. Fax: +886 2 23623040. E-mail addresses: [email protected], [email protected] (D.-J. Lee).
mixture as the solvent [12–17]. Extraction starts when solvent molecules penetrate the plant matrices, causing the cytoplasm layer to be exposed directly to the solvent, bringing about the dissolution of the bioactive compounds [18]. The mixture of ethanol and water at different concentrations is used as an extraction solvent in the extraction of bioactive compounds from plants [19–21]. The ethanol concentration affects the dielectric constant of the mixture hence altering the energy required to deteriorate the attraction of water molecules to the target molecules. The extraction temperature is also a critical factor to the stability of bioactive compounds and the extraction performance. High temperature is preferred for extraction of thermally stable compounds from plants since high temperature leads to an increase of the solute diffusivity and decrease of the energy barrier [22]. Conversely, the temperature of extraction should be limited for extracting thermal sensitive compounds. Liquid–solid ratio is also an important extraction parameter and when applied correctly can decrease the mass transfer barrier during the diffusion of active compounds and subsequently enhance the extraction yield [23]. However, if the ratio is beyond the optimum, the excess solvent does not have a significant effect on the equilibrium extraction yields and would lead to a solvent waste.
http://dx.doi.org/10.1016/j.jtice.2016.10.010 1876-1070/© 2016 Taiwan Institute of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010
ARTICLE IN PRESS
JID: JTICE 2
[m5G;December 5, 2016;15:52]
P.-S. Chiang et al. / Journal of the Taiwan Institute of Chemical Engineers 000 (2016) 1–8
Compressional puffing is a fast and inexpensive method for drying fruits and vegetables [24], and the process is carried out in a continuous puffing system at elevated pressure and in a superheated steam, which rapidly brings the water within the samples to a temperature above its atmospheric boiling point. When the samples are immediately exposed to atmospheric pressure, the intratissue vapor flashes to form a porous structure of plant biomass [25]. Huang et al. [26] introduced the process for bioactive compounds extraction due to the decomposition of the cellular structure of plant during the process. Also, comparing the other pretreatments such as ion-liquid, ultrasonic and microwave-assisted pretreatment, compressional-puffing provides advantages including relative simple procedure, reactant-saving, reduced pollution and feasibility for continuous production, while having great enhancement in extraction yield. P. morrisonicola products are commercially available which are based on time-consuming, ill-controlled biological hydrolysis reactions. The present study is the first report to note the optimal extraction parameter sets for hydrolyzed pine needles using compressional-puffing pretreatment. Since the puffing process shortens the treatment time from years to 10 s, the tested process is highly promising for yielding cost-effective and well-controlled digestate product by pine needle. The optimum extraction conditions for the phenolic compounds from pine needles were determined by using response surface methodology with a centralcomposite design. The effects of ethanol concentration, extraction temperature and liquid–solid ratio on the extraction yield (total flavonoids content and total phenolic content) and antioxidant activity (2,2 -diphenyl-1-picryhydrazyl (DPPH) scavenging activity, ferric reducing antioxidant power (FRAP) and 2,2 -Azino-bis(3ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) scavenging activity) are investigated. A quadratic model afforded a superior fit of the experimental data with r2 > 0.98 and p value <0.01. The optimum conditions to maximize the extraction yields and antioxidant activity of the pine needles extracts are: 36.1% v/v of ethanol, 70 °C and 50 ml/g of liquid–solid ratio. In addition, the pine needles extracts are also compared with commercial enzyme products and show competitiveness in both total phenolic content and antioxidant activity. 2. Materials and methods 2.1. Materials
with puffing temperature 220 °C. The unit possessed three compression (to 5 kg/cm2 )–relaxation cycles with cycle time of 10 s. After the three cycles, the chamber was opened to induce puffing.
2.3. Solvent extraction tests 2.3.1. Experimental design The treated pine needles were extracted in different extraction environments involving three variables (extraction temperature, ethanol concentration and liquid–solid ratio). A three-factor central composite design (CCD) was used to identify the relationship existing between the response function and the extraction process variables, as well as to determine those conditions that optimized the extraction process of total extraction content and the antioxidant activity of the pine needle extracts. The independent variables studied in this experiment were ethanol concentration (X1 : 25–75%), extraction temperature (X2 : 50–80 °C), and liquid–solid ratio (X3 : 23–57 ml/g), while response variables were total flavonoid content (TFC) (Y1 ), total phenol content (TPC) (Y2 ), DPPH radical scavenging activity (Y3 ), ferric reducing antioxidant power (Y4 ) and ABTS radical scavenging activity (Y5 ). The selection and range of these three factors was based on the premier experiment data. Each factor was coded at three levels (−1, 0 and 1) (Table S1). Twenty randomized experiments included six replicates as the center points were assigned based on CCD (Table 1).
2.3.2. Statistical analysis The Design Expert Version 10.0 (Stat-Ease, Minneapolis, MN, USA) software was used to conduct the experimental design and the statistical analysis. Results for the total content and antioxidant activity were expressed as means ± standard deviations. A response surface analysis and analysis of variance (ANOVA) were employed to determine the regression coefficients, statistical significance of the model terms and to fit the mathematical models of the experimental data that aimed to optimize the overall region for both response variables. The model proposed for the response is given below:
Yi = b0 + b1 X1 + b2 X2 + b3 X3 + b11 X12 + b22 X22 + b33 X32 + b12 X1 X2 + b13 X1 X3 + b23 X2 X3
(1)
The pine needles were collected from Nantou County, Taiwan. These needles were washed with tap water and then dried at room temperature to 6% w/w moisture content. The liquid samples from two commercial “enzyme products”, namely, Yamato Enzyme from Yamato Kouso Co., Ltd, Osaka, Japan and Gold Enzyme from Golden Enzyme, Nantou County, Taiwan, were used as the samples for comparison. Chemicals 6-Hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox), 2,2 -Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 2,2 -diphenyl-1-picryhydrazyl (DPPH), 2,4,6-Tripyridyl-s-Triazine (TPTZ), ferric chloride, formic acid (95%), gallic acid and iron(II) sulfate heptahydrate were from SigmaAldrich (St. Louis, USA) and acetonitrile was from Burdick and Jackson (New Jersey, USA). The Folin–Ciocalteu reagent was from Merck (Darmstadt, Germany). Quercetin was from Acros Organics (New Jersey, USA).
where Yi is predicted response, b0 is offset term that fixes the response at the central point; b1 , b2 and b3 are linear effect terms, b11 , b22 and b33 are the quadratic effect terms; and b12 , b23 and b13 are interaction effect terms. The proportion of variance explained by the polynomial models obtained is given by the multiple coefficients of determination, r2 , and the ANOVA analysis. The significance of each coefficient was determined using the Students t-test and p value, and nonsignificant coefficients were removed to obtain a reduced model. The optimum condition was verified by conducting experiments under these conditions. Responses were monitored and results compared with model predictions. The fitted polynomial equation was expressed as surface plots in order to visualize the relationship between the response and experimental levels of each factor and to deduce the optimum conditions.
2.2. The compressional-puffing treatment
2.3.3. Verification of the model Experimental data for the total content and antioxidant activity were obtained according to the recommended optimum conditions. The responses were determined after extraction of phenolic compounds under optimal conditions. The experimental and
The pine needle samples (<5 mm) were placed in compressional-puffing chamber MIBO R2 (Yuan Chuang Food Machinery Co. Ltd., Taiwan) [26]. And the pine needs were treated
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010
ARTICLE IN PRESS
JID: JTICE
[m5G;December 5, 2016;15:52]
P.-S. Chiang et al. / Journal of the Taiwan Institute of Chemical Engineers 000 (2016) 1–8
3
Table 1 Three-factor central composite design used for RSM with experimental and predicted values for the independent variables. Run
Coded factors X1
X2
Response X3
Y1 (TFC) exp
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 Run
−1.00 1.00 −1.00 −1.00 −1.00 1.00 1.00 1.00 0.00 0.00 0.00 0.00 −1.68 1.68 0.00 0.00 0.00 0.00 0.00 0.00
−1.00 −1.00 1.00 −1.00 1.00 −1.00 1.00 1.00 0.00 0.00 0.00 0.00 0.00 0.00 −1.68 1.68 0.00 0.00 0.00 0.00
Predicted
26.68 25.94 36.34 28.95 37.54 27.99 33.02 32.23 29.91 32.77 29.32 30.44 23.93 19.83 29.27 45.32 8.15 55.88 31.12 29.46
26.04 25.20 36.98 29.02 36.36 28.18 32.66 32.04 30.48 30.48 30.48 30.48 24.34 20.25 29.66 45.69 8.56 56.24 30.48 30.48
exp 29.79 28.17 39.70 33.28 43.09 31.50 32.90 35.54 32.50 31.83 29.91 30.40 24.64 18.81 28.65 45.39 8.34 61.56 32.50 31.10
Predicted 28.71 27.03 38.57 31.93 41.79 30.25 31.41 34.63 31.29 31.29 31.29 31.29 26.35 20.54 30.35 47.08 10.07 63.23 31.29 31.29
Response Y3 (DPPH)
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
−1.00 −1.00 −1.00 1.00 1.00 1.00 −1.00 1.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 −1.68 1.68 0.00 0.00
Y2 (TPC)
Y4 (FRAP)
Y5 (ABTS)
exp
Predicted
exp
Predicted
exp
96.6 85.5 112.5 123.0 139.5 95.6 103.4 98.1 105.6 118.1 113.2 117.7 99.0 59.8 111.9 152.2 30.5 206.1 121.9 115.9
95.4 83.5 132.7 102.5 139.7 90.6 90.4 97.5 103.3 103.3 103.3 103.3 100.8 55.3 112.1 149.1 15.6 191.1 103.3 103.3
241.2 227.1 243.7 267.6 283.5 243.1 251.3 247.2 240.3 253.8 250.1 251.4 246.0 188.0 247.9 365.5 87.8 387.4 259.7 249.8
242.1 230.5 245.4 266.8 286.9 244.9 250.6 248.1 251.0 251.0 251.0 251.0 244.2 186.2 246.0 363.4 86.1 385.4 251.0 251.0
4563 4263 6462 5844 7496 3781 5535 6020 4978 5103 5117 5089 4052 3188 5713 8194 1079 9742 5301 5213
predicted values were compared in order to determine the validity of the model.
2.4. Chemical analyses 2.4.1. TFC and TPC measurements The TFC was determined using the method of Park et al. [27] with modification. 0.5 ml of the extract solution was added to mixture of 5 ml ethanol, 0.25 ml 0.5 M sodium nitrite solution and 0.25 ml of 0.3 M aluminum chloride. Then, 1.5 ml of 1 M sodium hydroxide was added to the mixture. The absorbance of the mixture was measure at 510 nm with an UV–vis spectrophotometer (DR2700TM portable spectrophotometer, HACH, Colorado, USA) using quercetin as indicator. The TPC was measured using the method of Adefegha [28] with modification. 0.1 ml of the extract solution was added to mixture of 0.5 ml of Folin–Ciocalteu’s reagent, 5.9 ml deionized water. Then, 1.5 ml of 20% w/w sodium carbonate was added. The mixture was kept at 40 °C for 30 min. Then the absorbance of the mixture was
Predicted 4506 4196 6395 5787 7429 3714 5478 5963 5128 5128 5128 5128 4142 3279 5798 8277 1171 9825 5128 5128
measure at 765 nm with an UV–vis spectrophotometer using gallic acid as indicator. 2.4.2. Antioxidant activity assay The DPPH method was conducted for extract samples using the method proposed by Brand-Williams et al. [29]. 0.1 ml of example was added to 3.9 ml of the DPPH solution with vortexed for 1 min and left in the dark for 30 min at room temperature. The absorbance of samples was measured at 517 nm using an UV–vis spectrophotometer using Trolox as indicator. The FRAP potential of the samples was determined by the method of Benzie and Strain [30]. 1.5 ml FRAP solution (250 mM acetate buffer, 0.83 mM TPTZ (2,4,6-tripyridyl-s-traizine), 3.33 mM hydrochloride acid and 1.67 mM ferric chloride) was warmed to 37 °C and was mixed with 50 μl of the sample and 150 μl of the deionized water. The sample was incubated at 37 °C for 4 min and the absorbance at 593 nm using UV–vis spectrophotometer was recorded using ferrous sulfate as indicator. The ABTS radical scavenging activity for extract samples was determined based on the method of Arnao et al. [31]. 150 μl of the
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010
ARTICLE IN PRESS
JID: JTICE 4
[m5G;December 5, 2016;15:52]
P.-S. Chiang et al. / Journal of the Taiwan Institute of Chemical Engineers 000 (2016) 1–8 Table 2 Quadratic equations and statistical parameters calculated after implementation of central composite experimental design. Response
2nd order polynomial equation
R2
R2 (adjusted)
P value
Lack of fit
Y1 (TFC)
30.48 − 1.22X1 + 4.77X2 14.19X3 − 0.87X1 X2 −0.49X2 X3 − 2.9X1 X1 + 2.55X2 X2 + 0.68X3 X3 31.29 − 1.73X1 + 4.9X2 + 15.82X3 + 1.37X1 X2 −2.78X1 X1 + 2.32X2 X2 + 1.9X3 X3 103.34 − 13.54X1 + 11.03X2 + 52.21X3 + 2.25X1 X2 −9.84X1 X3 − 8.97X1 X1 + 9.65X2 X2 250.97 − 17.24X1 + 34.94X2 + 89.06X3 −6.79X1 X3 − 12.68X1 X1 + 19.04X2 X2 − 5.4X3 X3 5128.47 − 256.87X1 + 737.61X2 + 2575.53X3 −289X1 X3 + 90X2 X3 − 502.25X1 X1 + 676.32X2 X2 + 130.79X3 X3
0.9938
0.9833
<0.01
0.1963
0.9839
0.9596
<0.01
0.1223
0.9713
0.9312
0.0248
0.0496
0.9964
0.9885
<0.01
0.3903
0.9978
0.9930
<0.01
0.1542
Y2 (TPC) Y3 (DPPH) Y4 (FRAP) Y5 (ABTS)
extracts were reacted with 2850 μl of the ABTS solution at dark for 2 h. Then the absorbance was taken at 734 nm using the spectrophotometer using Trolox as indicator. All determinations were estimated in triplicate. 2.4.3. Other analyses Chemical composition of the phenolic compounds extracted from P. morrisonicola was analyzed by liquid chromatography–mass spectrometry (LC–MS) method. Chromatographic separation was performed by using the following binary gradient of two mobile phases, (A): 0.1% (v/v) aqueous formic acid and (B): acetonitrile consisting of 0.1% (v/v) formic acid, at a flow rate of 300 μl/min: 10% B (0 min), 10% B (10 min), 15% (20 min), 40% B (35 min). The UV detector was set at the wavelength of 280 nm with column temperature at 25 °C and the injection volume at 20 μl for all samples. The identifications of the active functional groups presence in the extracts from P. morrisonicola is conducted by FTIR spectroscopy (Perkin Elmer Spectrum 20 0 0, USA) over the wavelength range of 40 0 0–60 0 cm−1 . 3. Results and discussion 3.1. Extract characteristics Extract from P. morrisonicola, HPLC chromatograms and FTIR spectrums were conducted in order to characterize the components of extracted phenolic compounds. The HPLC chromatogram of pine needles extract and that of pure (+)-catechin hydrate were presented in Fig. S1, showing that both exhibited as major peak at 12 min. The FTIR spectra for the pine needle extract and for (+)-catechin hydrate were shown in Fig. S2. Based on these analyses, the present pine needle extract contain catechinlike substances enriched with O–H bond (3339 cm−1 ), C=O bond (1694 cm−1 ), aromatic ring (1603, 1516 and 1451 cm−1 ), and C–O–C bond (1257 cm−1 ). The pine needle extract samples were sent to SGS Taiwan Co. Ltd for characterizing quantities of 311 pesticide residues and that of copper (Cu), arsenic (As), lead (Pb), mercury (Hg) or cadmium (Cd). The results revealed that the extracts were free of these pesticide residues or heavy metals (Supplementary materials) and is regarded safe for human consumption. 3.2. Optimization of extract yields and antioxidant activities 3.2.1. Model fitting Experimental values of extraction yield (TFC and TPC) and antioxidant activity (DPPH scavenging activity, FRAP and ABTS scavenging activity) were used in a multiple regression analysis using response surface methodology to fit the quadratic equations (Eq. (1)). In this study, all experimental data were obtained from a 20-run-experiment with the pine needles pretreated with 220 °C
compressional-puffing, and predicted data were from a response surface analysis model with the results shown in Table 1. An ANOVA analysis for the quadratic model was used to test the significance and accuracy of the model. Highly significant levels for these (p < 0.01) were obtained by statistical analysis, while insignificant variables were ignored in order to get a precise and applicable model. The model, established by the regression equations, replaces the experimental real point to explain the response results. The regression equations, coefficients of determination (r2 ), adjusted r2 values, probability values (p) and lack of fit values for dependent variables are shown in Table 2. The results suggested that the extraction yield and antioxidant activity were primarily determined by the linear and quadratic terms. In addition, the predicted results according to the models were close to the observed experimental responses, indicating that the generated models adequately explained the data variation and can represent the actual relationships between the selected variables. 3.2.2. Effect of extraction parameters on extraction yield The data shown in Table 1 indicate that the extraction yield (total flavonoids content and total phenolic content) of the pine needles extracts and the extraction parameters were quadratic with a good coefficient (r2 = 0.9833 and 0.9596, respectively). The effect of ethanol concentration and extraction temperature on the total flavonoids content (TFC) is shown (Fig. 1a). The extraction yield increases with extraction temperature with a fixed ethanol concentration. High extraction temperature could lead to an increase in yield of the phenolic compounds through increasing phenolic solubility, decreased solvent viscosity and surface tension. Also the experiment was conducted between 50 and 70 °C since at higher temperatures there is degradation and ionization of anthocyanin which leads to a decrease in extraction yield and antioxidant activity [32]. The linear trend of the model with respect to extraction temperature is expected (Fig. 1c) The effect of ethanol concentration on extraction yield shows a quadratic fitting as a high peak is realized in the response surface of the effect of ethanol concentration and extraction temperature (Fig. 1a) and the effect of ethanol concentration and liquid–solid ratio (Fig. 1b). It is also in good agreement with previous section in other studies [33]. The effect of liquid–solid ratio is shown according to the response surface plots (Fig. 1b and c). Due to the fact that more solvent can enter plant cells while more flavonoids can permeate into the solvent under the higher liquid–solid ratio, a high ratio leads to greater extraction yield. The response of total phenolic content (TPC) shown (Fig. 1d–f) indicate a similar tendency and result with response to TFC. In conclusion, in order to maximize the extraction yield from P. morrisconicola, 70 °C extraction temperature, 50 ml/g of liquid–solid ratio and a moderate ethanol concentration should be adopted. 3.2.3. Effects of extraction parameters on antioxidant activities The data shown in Table 2 indicates that antioxidant activities (DPPH scavenging ability, FRAP and ABTS scavenging ability) of the
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010
JID: JTICE
ARTICLE IN PRESS
[m5G;December 5, 2016;15:52]
P.-S. Chiang et al. / Journal of the Taiwan Institute of Chemical Engineers 000 (2016) 1–8
5
Fig. 1. Response surface plot for the parameters considered. (a) Temperature and ethanol concentration on TFC; (b) liquid–solid ratio and ethanol concentration on TFC; (c) liquid–solid ratio and temperature on TFC; (d) temperature and ethanol concentration on TPC; (e) liquid–solid ratio and ethanol concentration on TPC; (f) liquid–solid ratio and temperature on TPC.
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010
JID: JTICE 6
ARTICLE IN PRESS
[m5G;December 5, 2016;15:52]
P.-S. Chiang et al. / Journal of the Taiwan Institute of Chemical Engineers 000 (2016) 1–8
Fig. 2. Response surface plot for the parameters considered. (a) Temperature and ethanol concentration on DPPH scavenging activity; (b) liquid–solid ratio and ethanol concentration on DPPH scavenging activity; (c) liquid–solid ratio and temperature on DPPH scavenging activity; (d) temperature and ethanol concentration on FRAP; (e) liquid–solid ratio and ethanol concentration on FRAP; (f) liquid–solid ratio and temperature on FRAP; (g) temperature and ethanol concentration on ABTS scavenging activity; (h) liquid–solid ratio and ethanol concentration on ABTS scavenging activity; (i) liquid–solid ratio and temperature on ABTS scavenging activity.
pine needles extracts and the extraction parameters are quadratic. The response surface plots are shown (Fig. 2a and i), and they show a great similarity with the results of extraction yield. Antioxidant activity of the pine needles extract appears to have the highest response in the range of 40–50% v/v ethanol concentration with respect to different extraction parameters. Extraction temperature causes a linear increase in the antioxidant activity which is similar to the result of extraction yield. Liyana-Pathirana and Shahidi [34] reported that the extraction rate of thermally stable antioxidants at elevated temperature was higher than the decomposition of less soluble antioxidants, which could be confirmed in the present study. The chemical degradation and decomposition during the extraction is the main mechanism leading to the decrease in extraction yield as well as antioxidant activity [35], therefore, a proper setting of extraction temperature could result in a maximum response, reported as 70 °C in this study. Liquid-to-solid ratio effects on the antioxidant activity of the pine needles extracts are much more significant than other extraction parameters due to the result of extraction yield [36] (see Table 3). 3.2.4. Verification of the models and optimization of extraction yield and antioxidant activity Table 1 lists that the experimental values were close to the predicted values. The optimal conditions obtained using the models
were as follows: 36.1% v/v ethanol concentration, 70 °C extraction temperature and 50 ml/g liquid–solid ratio. Under optimal conditions, the model predicted a maximum response of 39.6 quercetin equivalent (QE) mg/g DW of TFC, 44.1 gallic acid equivalent (GAE) mg/g DW of TPC, 151.8 Trolox equivalent (TE) μmol/g DW of DPPH scavenging activity, 315.5 mmol FeSO4 /g DW of FRAP and 7464 TEAC mg/g DW of ABTS scavenging activity. The extraction conducted at the optimal conditions yield: 38.4 ± 1.3 QE mg/g DW of TFC, 46.9 ± 0.7 GAE mg/g DW of TPC, 167.2 ± 3.3 TE μmol/g DW of DPPH scavenging activity, 309.9 ± 7.8 mmol FeSO4 /g DW of FRAP and 7277 ± 102 TEAC mg/g DW of ABTS scavenging activity. The experimental results correlate well with the model prediction. 3.2.5. Comparison with commercial products The extract from pine needs at the optimal extraction conditions (36.1% v/v aqueous ethanol, 70 °C and 50 ml/g of liquid–solid ratio) from 220 °C-puffing pretreated pine needles was compared its TPC, TFC and DPPH scavenging activity with those of the extract from original pine needles and from the two commercial products. At optimal extraction conditions, the puffing pretreatment enhanced TFC from 0.154 to 0.768 QE mg/ml, TPC from 0.211 to 0.938 GAE mg/ml, and DPPH scavenging activity from 1264 to 3344 TE μM. The Yamato Enzyme and Golden Enzyme contain no TFC but have 0.716 and 0.899 GAW mg/ml of TPC, respectively. The
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010
ARTICLE IN PRESS
JID: JTICE
[m5G;December 5, 2016;15:52]
P.-S. Chiang et al. / Journal of the Taiwan Institute of Chemical Engineers 000 (2016) 1–8
7
Table 3 Comparison of pine needles extracts with commercial products on phenolic compounds contents and antioxidant activity.
Untreated pine needles 220 °C-puffing pretreated pine needles Yamamoto enzyme (from fruit and vegetables with 60-d processing time) Golden enzyme (from fruit and vegetables with 90-d processing time)
corresponding DPPH scavenging activity data are 3198 and 2763 TE μM, respectively. Restated, the present pretreated-extracted sample has comparable quantity of TPC and antioxidant activity as the two commercial products. It is worth noting that the two tested commercial products were made from fruit and vegetables at a few months processing time. Conversely, the present sample is from pine needles, a biomass that are not consumable by human beings, unlike the fruit and vegetables adopted by the commercial products. Additionally, the total processing time for the present samples is within 24 h, much shorter than the long fermentation times needed for the commercial enzyme products. The tested process can yield extract with high TFC/TPC quantities of high antioxidant activities with short processing time.
4. Conclusions The optimal extraction parameters for puffing-pretreated P. morrisonicola to yield maximum PTC and PFC equivalents and autioxidant activities were identified using response surface methodology. Under optimum conditions (70 °C extraction temperature, 50 ml/g liquid–solid ratio and 36.1% v/v ethanol concentration), the extract has the following readings: 38.4 ± 1.3 QE mg/g DW of TFC, 46.9 ± 0.7 GAE mg/g DW of TPC, 167.2 ± 3.3 TE μmol/g DW of DPPH scavenging activity, 309.9 ± 7.8 mmol FeSO4 /g DW of FRAP and 7277 ± 102 TEAC mg/g DW of ABTS scavenging activity. The present extract from pine needles was noted to have comparable TPC equivalent and DPPH scavenging activities as noted from commercial products derived from fruit and vegetables.
Supplementary materials Supplementary material associated with this article can be found, in the online version, at doi:10.1016/j.jtice.2016.10.010. References [1] Kong ZM, Liu ZT, Ding BY. Study on the antimutagenic effect of pine needle extract. Mutation Res Lett 1995;347:101–4. [2] Yen GC, Duh PD, Huang DW, Hsu CL, Fu TYC. Protective effect of pine (Pinus morrisonicola Hay.) needle on LDL oxidation and its anti-inflammatory action by modulation of iNOS and COX-2 expression in LPS-stimulated RAW 264.7 macrophages. Food Chem Toxicol 2008;46:175–85. ˇ [3] Schäfer A, Chovanová Z, Muchová J, Sumegová K, Liptáková A, Dura cˇ ková Z, et al. Inhibition of COX-1 and COX-2 activity by plasma of human volunteers after ingestion of French maritime pine bark extract (Pycnogenol). Biomed Pharm 2006;60:5–9. [4] Yan JK, Wang YY, Ma HL, Wang ZB, Pei JJ. Structural characteristics and antioxidant activity in vivo of a polysaccharide isolated from Phellinus lintenus mycelia. J Taiwan Inst Chem Eng 2016;65:110–17. [5] Li K, Yang X, Hu X, Han C, Lei Z, Zhang Z. In vitro antioxidant, immunomodulatory and anticancer activities of two fractions of aqueous extract from Helicteres angustifolia L. root. J Taiwan Inst Chem Eng 2016;61:75–82. [6] Paun G, Neagu E, Albu C, Moroeanu V, Radu G. Antioxidant activity and inhibitory effect of polyphenolic-rich extract from Betonica officinalis and Impatiens noli-tangera berbs on key enzyme linked to type 2 diabetes. J Taiwan Inst Chem Eng 2016;60:1–7. [7] Heim KE, Tagliaferro AR, Bobilya DJ. Flavonoid antioxidants: chemistry, metabolism and structure-activity relationships. J Nutr Biochem 2002;13:572–84.
TFC (QE mg/ml)
TPC (GAE mg/ml)
DPPH scavenging activity (TE μM)
0.154 ± 0.009 0.768 ± 0.031 NA NA
0.211 ± 0.028 0.938 ± 0.014 0.716 ± 0.042 0.889 ± 0.013
1264 ± 74 3344 ± 165 3198 ± 312 2763 ± 103
[8] Parr AJ, Bolwell GP. Phenols in the plant and in man. The potential for possible nutritional enhancement of the diet by modifying the phenols content or profile. J Sci Food Agric 20 0 0;80:985–1012. [9] Hollman PCH, Katan MB. Dietary flavonoids: intake, health effects and bioavailability. Food Chem Toxicol 1999;37:937–42. [10] Sodeifian G, Sajadian SA, Ardestani NS. Evaluation of the response surface and hybrid artificial neutral network-genetic algorithm methodologies to determine extraction yield of Ferulago angulata through supercritical fluid. J Taiwan Inst Chem Eng 2016;60:166–73. [11] Zhu XW, Wei MP, Xu DP, Guo YH, Xie YF, Yao WR, et al. In vitro and in vivo antitumor effects of the extract of Sapindus spp. J Taiwan Inst Chem Eng 2016;66:27–32. [12] Touriño S, Selga A, Jiménez A, Juliá L, Lozano C, Lizárraga D, et al. Procyanidin fractions from pine (Pinus pinaster) bark: radical scavenging power in solution, antioxidant activity in emulsion, and antiproliferative effect in melanoma cells. J Agric Food Chem 2005;53:4728–35. [13] Kwak CS, Moon SC, Lee MS. Antioxidant, antimutagenic, and antitumor effects of pine needles (Pinus densiflora). Nutr Cancer 2006;56:162–71. [14] Bito T, Roy S, Sen CK, Packer L. Pine bark extract pycnogenol downregulates IFN-γ -induced adhesion of T cells to human keratinocytes by inhibiting inducible ICAM-1 expression. Free Radical Biol Med 20 0 0;28:219–27. [15] Ka MH, Choi EH, Chun AHS, Lee KG. Antioxidative activity of volatile extracts isolated from Angelica tenuissimae roots, peppermint leaves, pine needles, and sweet flag leaves. J Agric Food Chem 2005;53:4124–9. [16] Park YS, Jeon MH, Hwang HJ, Park MR, Lee SH, Kim SG, et al. Antioxidant activity and analysis of proanthocyanidins from pine (Pinus densiflora) needles. Nutr Res Pract 2011;5:281–7. [17] Zeng WC, Zhang Z, Gao H, Jia LR, He Q. Chemical composition, antioxidant, and antimicrobial activities of essential oil from pine needle (Cedrus deodara). J Food Sci 2012;77:824–9. [18] Rakotondramasy-Rabesiaka L, Havet JL, Porte C, Fauduet H. Solid–liquid extraction of protopine from Fumaria officinalis L. – Analysis determination, kinetic reaction and model building. Sep Purif Technol 2007;54:253–61. [19] Khiari Z, Makris DP, Kefalas P. An investigation on the recovery of antioxidant phenolics from onion solid wastes employing water/ethanol-based solvent systems. Food Bioprocess Technol 2009;2:337–43. [20] Durling N, Catchpole O, Grey J, Webby R, Mitchell K, Foo L, et al. Extraction of phenolics and essential oil from dried sage (Salvia officinalis) using ethanol–water mixtures. Food Chem 2007;101:1417–24. [21] Ofori-Boateng C, Lee KT, Saad B. A biorefinery concept for simultaneous recovery of cellulosic ethanol and phenolic compounds from oil palm fronds: Process optimization. Energy Conver Manag 2014;81:192–200. [22] Cissé M, Bohuon P, Sambe F, Kane C, Sakho M, Dornier M. Aqueous extraction of anthocyanins from Hibiscus sabdariffa: Experimental kinetics and modeling. J Food Eng 2012;109:16–21. [23] Franco D, Sineiro J, Pinelo M, Núñez MJ. Ethanolic extraction of Rosa rubiginosa soluble substances: Oil solubility equilibria and kinetic studies. J Food Eng 2007;79:150–7. [24] Kozempel MF, Sullivan JF, Craig JC, Konstance RP. Explosion puffing of fruits and vegetables. J Food Sci 1989;54:772–3. [25] Sullivan JF, Konstance RP, Aceto NC, Heiland WK, Craig JC. Continuous explosion-puffing of potatos. J Food Sci 1977;42:1462–3. [26] Huang CY, Wu SJ, Yang WN, Kuan AW, Chen CY. Antioxidant activities of crude extracts of fucoidan extracted from Sargassum glaucescens by a compressional-puffing-hydrothermal extraction process. Food Chem 2016;197:1121–9. [27] Park YS, Jung ST, Kang SG, Heo BG, Arancibia-Avila P, Toledo F. Antioxidants and proteins in ethylene-treated kiwifruits. Food Chem 2008;107:640–8. [28] Adefegha SA, Oboh G. Enhancement of total phenolics and antioxidant properties of some tropical green leafy vegetables by steam cooking. J Food Process Preserv 2011;35:615–22. [29] Brand-Williams W, Cuvelier ME, Berset C. Use of a free radical method to evaluate antioxidant activity. LWT Food Sci Technol 1995;28:25–30. [30] Benzie IFFF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of ‘Antioxidant Power’: The FRAP assay. Anal Biochem 1996;239:70–6. [31] Arnao MB, Cano A, Acosta M. The hydrophilic and lipophilic contribution to total antioxidant activity. Food Chem 2001;73:239–44. [32] Wang J, Sun B, Cao Y, Tian Y, Li X. Optimisation of ultrasound-assisted extraction of phenolic compounds from wheat bran. Food Chem 2008;106:804– 810. [33] Yang YC, Li J, Zu YG, Fu YJ, Luo M, Wu N, et al. Optimisation of microwave-assisted enzymatic extraction of corilagin and geraniin from Geranium sibiricum Linne and evaluation of antioxidant activity. Food Chem 20 0 0;122:373– 380.
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010
JID: JTICE 8
ARTICLE IN PRESS
[m5G;December 5, 2016;15:52]
P.-S. Chiang et al. / Journal of the Taiwan Institute of Chemical Engineers 000 (2016) 1–8
[34] Liyan-Pathirana C, Shahidi F. Optimization of extraction of phenolic compounds from wheat using response surface methodology. Food Chem 2005;93:47–56. [35] Pompeu DR, Silva EM, Rogez H. Optimisation of the solvent extraction of phenolic antioxidants from fruits of Euterpe oleracea using response surface methodology. Bioresour Technol 20 09;10 0:6076–82.
[36] Prasad KN, Hassan FA, Yang B, Kong KW, Ramanan RN, Azlan A, et al. Response surface optimisation for the extraction of phenolic compounds and antioxidant capacities of underutilised Mangifera pajang Kosterm peels. Food Chem 2011;128:1121–7.
Please cite this article as: P.-S. Chiang et al., Antioxidant phenolic compounds from P inus mor r isconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters, Journal of the Taiwan Institute of Chemical Engineers (2016), http://dx.doi.org/10.1016/j.jtice.2016.10.010