HIGH HYDROSTATIC PRESSURE: A METHOD WHICH INCREASES EFFICIENCY OF PHENOLIC COMPOUNDS EXTRACTION FROM FRUITS HAVING ADHESIVE NATURE E.M. ALTUNER (1)*, C. İŞLEK (2), T. ÇETER (1), H. ALPAS (3) (1) Kastamonu University, Faculty of Science and Arts, Department of Biology, Kuzeykent, TR-37100, Kastamonu, TURKEY, (2) Niğde University, Faculty of Science and Arts, Department of Biology, TR-51240, Niğde, TURKEY, (3) Middle East Technical University, Food Engineering Department, TR-06800, Ankara, TURKEY * Corresponding author:
[email protected]
ABSTRACT High Hydrostatic Pressure Processing (HHPP) has application areas other than food engineering, such as extraction of active ingredients from natural biomaterials. There are several studies about extracting phenolic compounds from substances having adhesive nature. In one of these studies extraction at room temperature, heat reflux and HHPP were used to extract flavonoids from propolis, where the extraction yield was 4.70±0.21, 4.56±0.11 and 5.10±0.14 (%w/w) respectively. In another study HHPP was used to extract phenolic compounds from Maclura pomifera fruits having very high adhesive nature and the results were compared with shaking at room temperature and soxhlet extraction. The highest amount of phenolic compounds extracted was 913.173 µg GAE/mL in 500 MPa by using solvent cocktail and the lowest amount was 316.877µg GAE/mL in soxhlet extraction by using methanol. As a result it could be concluded that HHPP can be a very effective extraction method for plant materials having adhesive nature.
1. INTRODUCTION High pressure is cold isostatic super high hydraulic pressure that ranges from 100 to 800 MPa or even more up to 1000 MPa (Altuner et al., 2006). High pressure extraction (HPE) is a novel non-thermal process which can be used effectively in order to extract active ingredients from plant materials (Zhang et al., 2005; Tokuşoğlu and Swanso, 2011; Altuner et al., 2012a; Altuner et al., 2012b). HPE is proven to be fast and more effective than most of other extraction methods (Zhang et al., 2004; US Food and Drug Administration Center [USFDAC], 2011, Altuner et al.; 2012b). This is true since pressurized cells show increased permeability due to the mass transfer theory (Dornenburg and Knorr, 1993; Ahmed and Ramaswamy, 2006; Zhang et al., 2005; Yan, 2002; Altuner et al., 2012b). High pressure increases the rate of dissolution. Since there is large differential pressure between the cell interior and the exterior of cell membranes, a rapid permeation is observed under HPE (Zhang et al., 2005). This increases the solvent penetration through the broken membranes into cells or increases the mass transfer rate due to increased permeability (Shouqin et al., 2004), which means the higher the hydrostatic pressure is, the more solvent can enter into the cell. As they enter, the more compounds can permeate the cell membrane which could cause the higher yield of extraction. The aim of this study is to put forward how high hydrostatic pressure processing increases efficiency of phenolic compounds extraction from fruits having adhesive nature.
2. METHODOLOGY There are several studies about extraction of phenolic compounds from substances having adhesive nature. Extraction of active substances some plant materials are not very easy because of their adhesive nature. Especially as the extracts stick inside the extraction medium such as Eppendorf tubes, huge amounts of active substance lost, which cause low amount of extraction yield. In order to get high amount of extraction yield of phenolic compounds, scientists try different types of extraction methods. HPE is one of those extraction methods (Altuner et al., 2012b). Although propolis is not a plant material, it is a strongly adhesive resinous biomaterial (Shouqin et al., 2004). According to some previous studies at least 200 compounds have been identified in propolis such as phenolic acids, flavonoids, phenolic esters, terpenes, and etc (Walker and Crane, 1987; Greenway et al., 1991; Bankova et al., 2000). There are some reports about the extraction methods of flavonoids from propolis (Paul and Morita, 1971; Richard, 1992; Chen and Zhou, 1996; Krell, 1998; Noble, 1998; Peter et al., 1998; Han et al., 2000; USFDAC, 2000; Gu et al., 2001; Lin et al., 2002; Murad et al., 2002; Yan, 2002), such as extraction at room temperature, heat reflux extraction and HPE (Shouqin et al., 2005). Shouqin et al. (2005) investigated various experimental conditions of the HPE, such as solvents, ethanol concentration (35-95%, v/v), HPE pressure (100-600 MPa), HPE time (1-10 min) and solid/liquid ratio (1:5-1:45 gcm-3), to optimize the extraction process.
On the other hand, Maclura pomifera fruit is a plant material having adhesive nature. Therefore, extraction of active substances from M. pomifera fruit is not very easy too. Altuner et al. (2012b) compared extraction of phenolic compounds by HPE, shaking at room temperature and soxhlet extraction. In this study, samples were pressurized at 250 and 500 MPa at room temperature for 10 min. Methanol (MeOH) and a solvent cocktail (dH2O:ethanol:methanol:acetone:CH2Cl2 - 1:2.5:2.5:2:2) were chosen as extraction solvents. Total phenolic content was expressed as µg Gallic Acid Equivalence/mL.
3. RESULTS AND DISCUSSION As a result of the study conducted by Shouqin et al. (2005), it was observed that the extraction yield of flavonoids from propolis was 4.70±0.21 (%w/w) for extraction at room temperature, 4.56±0.11 (%w/w) for heat reflux and 5.10±0.14 (%w/w) for HPE. Thus, the extraction yield with HPE for 1 min was higher than extraction at room temperature for 7 days and heat reflux extraction for 4 h respectively. Altuner et al. (2012b) found that the highest amount of phenolic compounds extracted was 913.173 µg GAE/mL by 500 MPa HHPP by using solvent cocktail and the lowest amount of phenolic compounds extracted was 316.877µg GAE/mL by soxhlet extraction by using MeOH. All methods were put in order from the highest amount to the lowest amount of extraction yield as 500 MPa HHPP - solvent cocktail, 500 MPa HHPP - MeOH, 250 MPa HHPP - solvent cocktail, shaking at room temperature with solvent cocktail, 250 MPa HHPP - MeOH, soxhlet extraction with solvent cocktail, shaking at room temperature with MeOH and soxhlet extraction with MeOH.
4. CONCLUSION As a result it can be concluded that HPE can be selected as a very effective extraction method for any biomaterials having adhesive nature such as propolis or M. pomifera fruit.
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