J.

of Supercritical Fluids 59 (2011) 5360

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The Journal of Supercritical Fluids

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Extraction of caffeine from Robusta coffee (Coffea canephora var. Robusta) husks using supercritical carbon dioxide
J. Tello, M. Viguera, L. Calvo
Department of Chemical Engineering, Universidad Complutense de Madrid, 28040 Madrid, Spain

a r t i c l e

i n f o

a b s t r a c t
This work evaluated the technical feasibility of supercritical CO2 extraction of caffeine from coffee husks, an abundant residue of the coffee industry. Different pre-treatments (initial humidity and milling) and operational conditions (pressure, temperature, time and ow rate) were studied in a CO2 continuous ow laboratory-scale unit. While prior wetting of the coffee husks was needed, milling was not required to extract the caffeine. The use of higher ow rates and/or operational times resulted in higher extraction rates. The process was favoured with increased operational pressure and temperature due to higher solubility. The maximum extraction yield obtained of this alkaloid was 84% when working at 373 K and 300 bar, using 197 kg CO2 /kg husks. After water washing, the caffeine was at least 94% pure. Comparing world production data, the initial caffeine content and global extraction yield data of other natural sources, this process could be very advantageous for its technological application. 2011 Elsevier B.V. All rights reserved.

Article history: Received 15 June 2011 Received in revised form 22 July 2011 Accepted 22 July 2011 Keywords: Caffeine Extraction Coffee husks Supercritical CO2

1. Introduction Caffeine (1,3,7-trimethylxanthine) is an alkaloid of the xanthine group widely known worldwide due to its occurrence in extensively consumed beverages, drinks and food. Natural sources of caffeine include different varieties of coffee beans (Coffea canephora, Coffea arabica), tea leaves (Camellia sinensis), guaran seeds (Paullinia cupana), mat leaves (Ilex paraguariensis), kola nut seeds (Cola nitida, Cola acuminata) and cocoa beans (Theobroma cacao) [1]. Despite the fact that tea is globally consumed more widely than coffee, coffee is the main source of caffeine in daily consumption given its generally higher caffeine content [2]. Other dietary factors that contribute to daily overall caffeine consumption include foods like certain soft and energy drinks, chocolate, candies and sweets, as well as that contained in some medications [3], such as stimulants, diet aids, painkillers and cold remedies [4]. Numerous studies have reported the effects of caffeine consumption in humans, such as the well-known stimulant effect of low doses of caffeine on the nervous system, which enhances concentration capacity and counteracts tiredness. However, in more sensitive individuals (or under bigger doses of caffeine), this effect could generate episodes of insomnia, anxiety, nervousness, irritability, hostility and mood swings. Some other reported

Corresponding author. Tel.: +34 913944185. E-mail address: lcalvo@quim.ucm.es (L. Calvo). 0896-8446/$ see front matter 2011 Elsevier B.V. All rights reserved. doi:10.1016/j.supu.2011.07.018

physiological effects are stimulation of the gastric and urinary systems and increased heart rate and blood pressure [48]. These negative effects have led to an increasing consumption of decaffeinated coffee, generating the development of various processes that remove this alkaloid from coffee beans. Among these processes, organic solvents with high toxicity (methylene chloride, ethyl acetate), Swiss water decaffeination (which results in a less avourful brew than other methods) and the employment of carbon dioxide in supercritical conditions are commonly used [9]. The latter was successfully developed on an industrial scale in the 1970s, based on Kurt Zosels patent [10], initially run by Cafe HAG and General Foods [9]. Compared with other conventional methods, this process showed better results, both in terms of generating a higher quality product and for being a better process from an environmental point of view. CO2 is a non-ammable and non-toxic solvent that can be easily removed from the nal product. These advantages have driven the subsequent investigation of supercritical extraction of caffeine from other natural sources, such as tea leaves [11], stalks and bre wastes from industrial tea processing [12,13], guaran seeds [14,15], mat leaves [15], cocoa beans [15,16] and coffee oil [17]. On the other hand, the low solubility of xanthines in supercritical CO2 , due to its apolar character, makes the addition of polar cosolvents (such as water or ethanol [13,15,1820]) to the compressed gas an interesting practice. Cosolvent effects are due to specic chemical (hydrogen bonds and acidbase interactions) or physical interactions (dipoledipole or dipole-induced dipole) between the cosolvent and solute, and also to a possible interaction between the solvent and the cosolvent, affecting solvent-solute

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Fig. 1. High pressure apparatus used for the supercritical uid extraction of caffeine from coffee husks. PI, pressure indicator; TI, temperatura indicator; TIC, temperature indicator and controller; TC, temperature controller; BPR, back pressure regulator.

interactions. These effects result in accelerating and making the extraction easier [15]. Regarding the operational conditions, the solvent power of supercritical CO2 is highly dependent on pressure and temperature, variables that can be independently modied. Thus, different authors have reported higher yields when working at high pressures, due to the enhanced CO2 solvent power, and when working at higher temperatures [14,16,2022]. Among all the natural sources previously cited, coffee could be the most important worldwide from an economical point of view. FAO estimated its world production in 2009 at more than eight million tons [23]. In order to be used and consumed, green coffee is processed by two different methods, the wet or dry method. On average, for every ton of clean coffee produced, 1 ton of husks are generated during dry processing, whereas, for wet processing, 0.28 tons of parchment husks, 2 tons of pulp and 22.73 tons of wastewater are generated [24]. Obviously, this great amount of residues represents a serious source of pollution and environmental problems. Recent efforts to solve this problem include activities like the production of organic fertilisers [25], using these residues as a substrate for the synthesis of biomass [26] and bio-ethanol [27], for the production of a diversity of products through fermentation (citric acid [28], pectins, pectic enzymes and vinegar [29]), as a natural source of anthocyanins [30], for the growth of mushrooms, as an adsorbent [27] and as a source of caffeine by percolation with alcohols and water [31]. Alternatively, coffee husks have been used for animal feed, because of their high cellulose content, which can be metabolised by ruminants. However, given its structure and its high content of lignin, silica [31], polyphenols, tannins (5% [26]) and caffeine (1.3% [26]) [32], this use is limited. Thus, the most common nal use for husks is in coffee mills to provide the energy needed for the nal drying of the coffee beans [31]. The aim of this work was to evaluate the feasibility of extracting caffeine from coffee husks using carbon dioxide at supercritical conditions, as a means of using this residue to obtain high-value caffeine that may be subsequently used in the food, pharmaceutical, veterinary or cosmetic industries.

2. Materials and methods 2.1. Materials Whole coffee husks of C. canephora var. Robusta were obtained from Manaus (Brazil). They were previously characterised in Natraceutical group by standard methods, with a fat content of 1.0%, a moisture content of 16.4% and a caffeine concentration of 1.1%. This material was stored in dry conditions at room temperature during the investigation. Pressurised liquid carbon dioxide (99.998% in purity) was kindly donated by Carburos Metlicos S.A. Caffeine, 99.99% in purity and used in the solubility experiments, was provided by the Natraceutical Group. The packing agents were round perforated plastic beads, with an average diameter of 2 mm. They were also used to create the inert bed in the net caffeine solubility measurement experiment. Redistributors were made from a stainless steel mesh with same area as the cross section of the extractor. 2.2. Pre-treatment To evaluate the inuence of the condition of the raw material on the extraction rate, two different pre-treatments were performed. First, coffee husks were ground by the commercial stainless steel blades of a coffee grinder. Second, the material (raw or ground) was humidied to the desired percentage of humidity (measured by weight) with ultrapure water. Thus, the following pre-treatments were assayed: 1. 2. 3. 4. 5. 6. Whole coffee husks with a natural humidity of 16%. Whole coffee husks with a humidity of 32%. Whole coffee husks with a humidity of 48%. Whole coffee husks with a humidity of 64%. Ground coffee husks (32% humidity) loaded with redistributors. Ground coffee husks (32% humidity) loaded with redistributors and packing agents (36%, w/w).

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7. Ground coffee husks (32% humidity) loaded with redistributors and packing agents (50% w/w). 2.3. Apparatus description and extraction process Supercritical uid extraction was conducted using the laboratory scale system previously described by Calvo et al. [33], consisting of a CO2 feeding line, a 50 mL capacity 316 ss extractor, a pressure control device and a sample collection system, as shown in Fig. 1. The CO2 was fed from the pressured container in the liquid phase, then cooled in a temperature-controlled bath (Selecta, Frigiterm-30) to 258 K and impelled by a cooled-head membrane pump (Milroyal D, Dosapro Milton Roy). This refrigeration prevented pump cavitation in the pressurisation process. The pressurised CO2 is pre-heated in a coil located inside a heating jacket prior to entering the vessel. The vessel is also thermally conditioned by another heating jacket; both with a temperature control of 1 K and recorded by a type K thermocouple placed inside the reactor in direct contact to the uid and the solid. Pressure was read by a Bourbon manometer with an accuracy of 5 bar. The control of the pressure and ow rate was achieved by both a heated micrometering valve (Tescom, Serie 26-1700) and the pump. A safety rupture disk set at 380 bar prohibited the pressure from exceeding the prescribed values. The amount of CO2 per unit of time was determined in a mass ow meter (Alicat Scientic, M-10SLPM-D) connected at the end of the line with an accuracy of 0.5 g/min. Coffee husks, previously weighed and adequately pre-treated, were loaded in the extractor, forming a x bed. When using whole husks 8.01 0.28 g were used. When using milled coffee husks, the quantity of 15.75 1.64 g was loaded into the extractor. Redistributors made of stainless steel mesh having the same size of the cross section of the extractor, were introduced approx every 1.5 cm in the tests done with the ground material. Then, the extractor was closed and pre-heated. After that, the CO2 was pumped in, and, once the desired pressure was reached, the back pressure regulator (BPR) was opened, providing a continuous ow through the bed. After the valve, the CO2 was depressurised, the solvent power of the supercritical uid dropped and the extract precipitated in a previously weighed glass ask. When the established treatment time was over, the apparatus was depressurised, the extract weighed and the coffee husks unloaded. The total amount of CO2 circulated was read by the mass ow meter. Since caffeine is a solid product, part of it stuck to the walls of the equipment. To recover it, after each experiment, a washing procedure was conducted. To do this, a portion of cotton was soaked in 40 mL of ultrapure water and loaded in the extractor. Then, CO2 at 323 K was own through the extractor at a pressure of about 60 bar (container pressure) with a ow rate of 23 g/min for 20 min. The procedure was repeated twice. The collected water with the corresponding dissolved caffeine was placed in an oven at 323 K. Once the water was evaporated, the precipitate was weighted, properly labelled and stored for its subsequent composition and purity analysis. Similar washing methods with polar solvents were previously [34]. used by Saldana 2.4. Analysis of caffeine content on the extracts The determination of caffeine in the extracts was performed by HPLC following the protocols stated by Ic en and Gr [12]. Briey, extracts were diluted with ultrapure hot water, ltered under vacuum, cooled and analyzed by means of an Agilent 1100 Series chromatograph equipped with an UVvisible detector set at 320 nm, and a C-18 column. The mobile phase used was acetonitrile (30%, v/v) (Merck) at 0.8 mL/min ow rate. Calibrations

were performed using standard caffeine solutions at different concentrations. 2.5. Measurement of caffeine solubility To measure the solubility of net caffeine in pure CO2 , a given amount of caffeine was charged in the extractor on an inert bed made of plastic beads. The rest of the operation was performed as described in Section 2.4. 2.6. Determination of results Extraction yield data were expressed as the percentage of total extract obtained with respect to the initial content of fats and caffeine. Extraction Yield (%) : Extract (g ) 100 [Fats (g ) + Caffeine (g )]initial

Caffeine yield data were expressed as the percentage of caffeine in the extracts (obtained from purity data) with respect to the initial content of caffeine. Caffeine Yield (%) : Extract (g ) Caffeine content (%) Initial Caffeine (g )

2.7. Experimental standard deviation To assess the reproducibility of the results, two experiments were repeated six times each, establishing an experimental standard deviation of 9% in extraction yield and 7% in caffeine purity. All data are the average of at least two experiments. 3. Results and discussion This section rst presents the effect of pre-treatment of the raw material on the yield of the extraction process. Thus, the initial moisture content and the particle size of the raw material were explored. The use of agents to prevent the agglomeration of the bed was then investigated to enhance contact. Regarding the variables that affect the extraction process, different values of pressure, temperature and operational times combined with different CO2 ow rates were studied. Caffeine extraction from coffee husks is shown as a curve representing the yield results versus the circulated CO2 mass. Caffeine solubility data were obtained to assess the outcome of our collection procedure, comparing our results with those previously reported by other authors [35,36]. Finally, a study of the composition of the extracts in terms of caffeine purity was done. 3.1. Caffeine solubility measurement In order to verify the reliability of the experimental data obtained with our apparatus, a measurement of the solubility of net caffeine in pure CO2 at 333 K and 200 bar was done, comparing the results obtained with those previously reported by Johannsen et al. [36]. The results are shown and Brunner [35] and Saldana in Fig. 2. Our result closely agreed, validating our extraction and washing procedure. Caffeine solubility showed a direct relationship with operation conditions. The higher the pressure was, the higher the amount of extracted caffeine. This result was attributed to the increased density of the solvent and, consequently, to its solvent power. On the contrary, the effect of temperature depended upon the working pressure, with retrograde behaviour at pressures lower than about 200 bar. This pressure, where the solubility isotherms cross,

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6 5

4 3 2 1 0 100

150

200

250

300

350

400

Pressure (bar)
Fig. 2. Solubility of caffeine in supercritical CO2 at different pressures and temperatures ( : 313 K, Johannsen and Brunner [35]; : 333 K, Johannsen and Brunner [35], et al. [36], 323 K, Saldana : 353 K, Johannsen and Brunner [35]; +: 313 K, Saldana et al. [36], and : 333 K, this work). et al. [36], : 343 K, Saldana

is known as the crossover pressure [37]. Below it, the caffeine solubility decreased as temperature increased. Above this value, an increase in temperature increased solubility [17]. It is well-known that this behaviour is a consequence of the opposing effects of the solvent density variation and the solute vapour pressure. Below the crossover pressure (about 200 bar), small increases in temperature result in drastic decreases in solvent density and consequently, on solvent capability. As an example, at 180 bar, caffeine solubility decreases from 1.16 to 0.87 g/kg CO2 when augmenting the temperature from 323 to 343 K [36]. As the process pressure moves away from the solvent crossover pressure, its density become less sensitive to temperature changes and the effect that prevails is the increase in vapour pressure. Thus, important growths on caffeine solubility are seen over 200 bar in the isotherms as the pressure increases. This change in the inuence of temperature may also be seen in the isotherms obtained with CO2 mixed with cosolvents, although in this case, the crossover pressure acquires a higher value. For example, for mixtures of CO2 isopropanol (5%) or CO2 ethanol (5%), the crossover pressure increases to 210 and 230 bar, respectively [37]. 3.2. Effect of coffee husks initial moisture content In industrial supercritical CO2 decaffeination processes, a previous pre-wetting of the coffee beans is needed in order to make the process feasible [9,38,39]. Similarly, we found that it was necessary to wet the coffee husks prior to extraction, since hardly any caffeine was removed when working with coffee husks as received, even at
30

the most extreme conditions allowed by the apparatus (350 bar and 373 K) (sample 1, Fig. 3). Knowing the initial moisture content of the coffee husks provided (16.4%), higher hydration values were tested, i.e. 32%, 48% and 64% (samples 2, 3 and 4, in Fig. 3, respectively). The results indicated that increasing the amount of water up to 32% resulted in higher extraction yields, but exceeding this percentage was not benecial. For example, when the water content was 64%, the extraction resulted in the removal of only 5% of the extractible compounds originally present in the husks. Moreover, this high moisture level generated a malfunction of the apparatus, with the formation of ice crystals in the outlet valve of the extractor, which interrupted the normal ow of CO2 . Because of this, coffee husks at 32% moisture were chosen as the starting material for the following experiments. In light of these results, it can be concluded that the hydration of coffee husks prior to the extraction process is a critical factor. This fact could be attributable to the hydrolytic rupture of hydrogen bonds which link the caffeine to the natural matrix on which it is adsorbed, liberating it to be swept out by the supercritical CO2 [39]. Furthermore, water could contribute to swelling of the cell membrane, leading to the enhancement of solute diffusion out of the plant tissues [11,40]. In fact, we observed an important volume augmentation of the husks after wetting. Another indirect reason could be the entrainer effect of the water dissolved by the CO2 during the extraction, facilitating the dissolution of polar compounds [40] such as caffeine. Supporting this hypothesis, Iwai et al. noticed that solubility of caffeine in supercritical CO2 was found to increase 22% when using CO2 with saturated water at 313.2 K and 150 bar [41]. Similarly, Park et al. found an important increase (from 9% to 75%) on caffeine extraction yield from green tea, when using CO2 wetted with 8.8% water [18]. The lower extraction yield obtained in the test conducted at 64% moisture was presumably due to the preference of the caffeine to remain soluble in the excess aqueous phase, as suggested by Pourmortazavi and Hajimirsadeghi [40]. 3.3. Effect of grinding and use of packing and redistributors In general, grinding is an operation that improves the extraction of compounds because it decreases the particle size, augmenting the contact area with the solvent [42,43]. However, it is a high energy cost operation, so it is important to evaluate the need for this procedure. In order to assess the impact of grinding the raw material, yields obtained at xed conditions in both whole and ground coffee husks were compared (samples 2 and 5, in Fig. 3, respectively). The results show that grinding did not improve the process performance, but also hindered the operation. This could be because milling the coffee husks together with the addition of water generated a very

Solubility (g/kg CO 2 )

Extraction Yield (%)

25 20 15 10 5 0

3 6 7 4 5

1
W (16,4% moisture) W (32% moisture) W (48% moisture) W (64% moisture) G (32% G (32% G (32% moisture) + R moisture) + R + moisture) + R + P (36%) P (50%)

Fig. 3. Effect of different pre-treatments on the extraction yields with supercritical CO2 at 200 bar and 333 K with a mass ratio of 24 g CO2 /g husks (sample 1 extracted at 350 bar and 373 K) (W, whole coffee husks; G, ground coffee husks; R, redistributors; P, packing).

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100 90

Table 1 Experimental conditions to obtain increasing amounts of CO2 for Fig. 4. CO2 ow (g/min) Time (min) 180 120 180 120 CO2 mass (g) 116 295 436 702

80

Extraction Yield (%)

70 60 50 40 30 20 10 0 0 200 400 600 800

0.6 2.5 2.4 5.8

CO2 mass (g)

Fig. 4. Comparison between the extraction curves obtained for pure caffeine ( ) and for the total extract from coffee husks (). Conditions: 333 K and 200 bar (broken line: linear tting; solid line: second grade polynomial tting).

saturated. (The curve was drawn based on the solubility value under the same operating conditions. This comparison demonstrates the difculty of caffeine extraction from coffee husks. For example, a quantity of 150 g of CO2 was enough to achieve total extraction of pure caffeine if it was completely accessible, while for coffee husks, this amount could only retrieve 15% of the extractible compounds (that is, a mixture of caffeine, fats and other minor compounds.) The low extraction yield achieved may be attributed to two reasons. First of all, it could be due to the capacity of caffeine to form complexes with certain compounds naturally present in coffee husks (such as chlorogenic acids) which would prevent its total removal [15,20] even in the presence of water. Second, it could be related to the strong resistance of the internal mass transfer. 3.5. Effect of operating conditions The density of a supercritical uid varies dramatically with operational pressure and temperature. Thus, one of the most characteristic properties of supercritical uids is the wide range of densities that can be attained depending on the pressure and temperature of work. Table 2 shows the estimated density values corresponding to the operational conditions used in this work [48]. Consequently, and due to the direct relationship between the density of a uid with its solvation capacity, gases at supercritical conditions can vary enormously in terms of solvent power by small variations of pressure and/or temperature. On the other hand, the change of these two variables generates signicant changes in CO2 transfer properties, such as viscosity and diffusivity, which therefore impact on the penetrability of CO2 into solid matrices. In general, increasing the operational pressure generates an increment of viscosity and a reduction of the selfdiffusion coefcient, while an increase in temperature causes the opposite effect. Finally, when it comes to plant materials, it is especially important to maintain the temperature in a certain range since it could alter the properties of both the natural source and the solutes being extracted. For all these reasons, when evaluating a supercritical uid extraction process, it is mandatory to study the impact of these two variables. 3.5.1. Effect of temperature on extraction yields As discussed in Section 3.1, the impact of temperature on the solubility of extractible compounds depends on the working
Table 2 Density of supercritical CO2 under the experimental conditions used in this work [48]. Pressure (bar) 60 100 150 200 250 300 300 300 300 300 300 300 Temperature (K) 373 373 373 373 373 373 363 353 343 333 323 313 Density (kg/m3 ) 100 189 333 481 589 662 703 746 788 830 870 910

compact bed, so CO2 would pass through preferential channels (probably around the bed) without making efcient contact with the raw material. This negative effect is common in beds made of nely ground raw materials [44]. To eliminate this negative effect, mesh was inserted together with the raw material at different levels in the xed bed to facilitate CO2 radial redistribution, as previously recommended [45]. We also studied mixing of the ground husks with a packing material consisting of inert perforated beads used in two different proportions by weight: 36% and 50% (samples 6 and 7 of Fig. 3, respectively). Its purpose was to increase the void fraction of the bed facilitating the CO2 ow and contact. Our results showed that the use of these agents improved the extraction yield compared to those samples in which they were not employed, but, owing to the fact that the results obtained were not better than those from the whole raw material, we opted to continue the investigation with coffee husks without grinding, which would mean important energy savings at the industrial scale. 3.4. Extraction curve The course of the extraction process can be followed by determining the amount of extract against the time of extraction or the solvent used. If the extractible compounds are easily accessible, the obtained curve presents two different parts; the rst one is a straight line (corresponding to a constant extraction rate) while in the second part, the curves bends down, approaching a limiting value which is given by the total amount of extractible substances [46]. In cases of a low initial concentration of extract in the solid substrate or an extract not readily available for the solvent, transport within the solid dominates from the beginning of the operation. In this case, the extraction curve does not show these two parts, but approaches an asymptote, given by the distribution coefcient corresponding to the initial concentration of the extract in the solvent [47]. Of the latter type are those obtained for the extraction of caffeine from coffee husks using supercritical CO2 . An example obtained at 333 K and 200 bar is given in Fig. 4. The extraction yield increased at an ever-decreasing rate as the mass of CO2 was increased. Increasing amounts of CO2 owing through the xed bed were achieved by increasing the CO2 ow rate and/or by using longer operational times (see Table 1). Fig. 4 also compares the extraction yield with the one that would result if all the caffeine initially contained in the coffee husks would be fully accessible, so compressed CO2 would leave the extractor

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40 35 Extraction Yield (%)

70 60 50 40 30 20 10

Extraction Yield (%)

30 25 20 15 10 5 303

0 0 50 100 150 200 250 300 350

323

343

363

383

Pressure (bar)
Fig. 6. Inuence of pressure on the extraction yield obtained from whole coffee husks at a xed temperature of 373 K, a xed water content of 32% and a total CO2 mass of 429 g ( : Experimental data; Curve line: second grade polynomial tting).

Temperature (K)
Fig. 5. Inuence of temperature on the extraction yield obtained from whole coffee husks at a xed pressure of 300 bar, a xed water content of 32% and a total CO2 mass of 320 g ( : experimental data; solid line: linear tting).

pressure, which depends on whether the operation is carried out above or below the crossover point. In Fig. 5, extraction yields from wet and whole coffee husks using CO2 at 300 bar with increasing temperature from 313 to 373 K are shown. The solvent to feed ratio was kept xed at 39 g CO2 /g (experimental runs were obtained at constant ow rates). Under the same treatment conditions, higher yields were obtained when working at higher temperatures, as was expected due to operating above the crossover point, leading to an enhancement in caffeine solubility. This fact has been previously reported by et al. [15], who concluded that the extraction yield of Saldana methylxanthines from various natural sources at 400 bar (above the crossover point) increased with increasing temperature, whereas when working at 100 bar, there was a retrograde behaviour. This result also agrees with those obtained by Kim et al. [11], whose caffeine extraction yields from green tea leaves increased from 54% to 66% by increasing the working temperature from 313 to 353 K while operating at 400 bar. Similarly, in the extraction of caffeine from [34] found that by increasing the working coffee beans, Saldana temperature from 313 to 343 K, the amount of material extracted when working at 280 bar increased, although much of the extract was the essential oil of coffee, which in green coffee beans of the Robusta variety comprises between 9 and 13% (w/w) of the raw material. Another reason for this behaviour is the increase in the mass transfer rate when increasing operational temperature, due to an improvement in the diffusion coefcient [49]. Knaff and Schlnder showed that for a range of only 10 K (from 323 to 333 K), the diffusion coefcient of caffeine in supercritical CO2 varied from 1.506 108 m2 /s to 1.992 108 m2 /s when working at a xed pressure of 137 bar [50].

Furthermore, the colour of the extracts was inuenced by the working temperature; thus, samples obtained at higher temperatures showed a dark brown colour instead of the yellowish-green seen in the samples obtained at lower temperatures. This colour transformation was possibly due to the browning of some of the pigments naturally present in the coffee husks. 3.5.2. Effect of pressure on extraction yields Pressure is another factor that sets the solvent capacity of supercritical CO2 . As discussed in Section 3.1, an increment of the working pressure causes an increase in the density of the compressed gas (see Table 3), which in turn enhances the gas solvent capacity for the removal of the caffeine. To check whether this trend was maintained in the extraction process, different pressure values were explored maintaining a xed temperature of 373 K and a solvent to feed ratio between the compressed gas and the coffee husks of 53 g/g. Results are shown in Fig. 6. Indeed, and as expected, an increase in the operating pressure exponentially increased the efciency of extraction. A raise in pressure from 100 to 300 bar, multiplied by ten the extraction yield. This et result is consistent with those found in literature. Thus, Saldana al. [15] conrmed the effect of variation of pressure on extraction yield: working at 100, 200 and 300 bar resulted in the removal of 0.26%, 60% and 96% of the initial caffeine present in guaran seeds, respectively. Moreover, Kim et al. [11] improved the yield of caffeine extracted from tea leaves from 3% to 28% when increasing the operational pressure from 200 to 400 bar working at a x temperature of 323 K using compressed CO2 modied with water (7%, w/w). Similarly, Ic en and Gr [12] reported increments on the extraction of caffeine from tea stalks from 13.6 to 14.7 mg/g when augmenting the pressure from 150 to 200 bar at 333 K.

Table 3 Total extraction and caffeine yields obtained under different conditions. T (K) 333 353 373 373 373 373 P (bar) 300 300 300 200 300 300 Solvent to raw material mass ratio (gCO2 /g) 35 36 40 53 58 197 Time (min) 120 105 120 100 105 300 Extract yield (%) 24 28 35 29 65 59 Purity of caffeine (%) 63 74 61 65 64 77 Caffeine yield (%) 27 39 40 36 78 84

J. Tello et al. / J. of Supercritical Fluids 59 (2011) 5360 Table 4 Conditions used and caffeine extraction yields from several natural sources obtained using pure supercritical CO2 . Raw material 2009 world productiona (MM tons) 4.2 N. A. 0.8 3.9 3.9 3.9 8.3 8.3 Initial caffeine (%) 0.60.8b 4.3b 0.80.9b 2.8b 1.16 0.92 1.1 1.1 Pressure (bar) 400 400 400 300 250 250 300 300 Temperature (K) 343 343 343 343 333 333 373 373 Solvent to raw material mass ratio (gCO2 /g) 43.7 399 1140 102 924 924 197 58 Yield (%) Ref.

59

Theobroma cacao (cocoa beans) Paullinia cupana (guaran seeds) Ilex paraguariensis (mat leaves) Camellia sinensis (tea leaves) Camellia sinensis (tea stalks) Camellia sinensis (bre wastes) Coffea canephora (coffee husks) Coffea canephora (coffee husks) N.A., Datum not available. a FAO Statistics [23]. b Data obtained from Ref. [1].

66 97 96 9 62 66 84 78

[16] [15] [15] [17] [12] [12] This work This work

3.6. Caffeine purity in the extracts Along with the caffeine extracted, other unidentied compounds naturally present in coffee husks were recovered, primarily fatty compounds. Table 3 shows the purity of selected extracts obtained under different conditions. The main component in all the extracts obtained was caffeine, with an average value of 67%. This high selectivity of supercritical CO2 for caffeine in coffee beans has been previously reported [34]. This value was not substantially altered when by Saldana varying experimental conditions. As a result, caffeine yields varied proportionally to total extraction yields. In other words, an increment in operational pressure, temperature and solvent to raw material mass ratio resulted in higher extraction rates, achieving the removal of more than 80% of the initial caffeine when working under the most extreme conditions. On the other hand, the rest of the compounds were easily removable by simple water washing, which generated two immiscible phases: a fatty one with the vast majority of the undesirable compounds, and an aqueous one in which the caffeine was selectively soluble. After evaporating this water, extracts had purity higher than 94%. 3.7. Comparison with other natural sources Table 4 shows a summary of the yields of caffeine extracted from different natural sources with pure supercritical CO2 . The operational conditions employed in the processes are also included. The highest extraction yield of caffeine obtained from coffee husks (about 84%) was a good value, even though it was not as high as those obtained in supercritical extraction from guaran seeds and mat leaves. However, compared to these other sources, coffee husks present further advantages as a raw material for the production of caffeine. First, the world production of green coffee is higher than that of the other natural sources cited, which generates (as commented in Section 1) a large quantity of husks rich in caffeine during the manufacturing process. Second, the procedure requires the use of much lower solvent to raw material mass ratios (60200 g CO2 /g). Consequently, this material is denitely an interesting option for obtaining caffeine on an industrial scale, as well as from an economical and environmental point of view. 4. Conclusions The present study evaluated the technical feasibility of extracting the caffeine naturally present in Robusta coffee husks using CO2 under supercritical conditions, as a novel use of this common and abundant residue from the coffee industry.

The results revealed that to make the process feasible, pretreatment was required, consisting of pre-wetting of the raw material up to 32% moisture, which is close to that needed for coffee beans (30%) and doubled the original amount of water present in the coffee husks. Grinding the coffee husks seemed to be a process that hindered the removal of the compound of interest, due to the strong compaction of the bed. It seems that coffee husks were porous enough to allow the circulation of the gas, easily extracting the available caffeine. On the other hand, the use of greater quantities of CO2 resulted in a large increase in the amounts of caffeine extracted, as the CO2 did not reach saturation. These greater amounts could be achieved either by increasing the CO2 ow rate or by increasing operational times. Optimum relationships should be deducted through an economical analysis. Regarding the operational conditions, it was observed that the more extreme the operational conditions were (pressure and temperature above the crossover point), the higher the extraction yields achieved. Pressure variation was more effective, because it affected the density of the supercritical uid and, therefore, its solvent capacity. Thus, 84% of the initial caffeine was extracted, after treating the material for ve hours with 197 g of compressed gas per g of raw material at 373 K and 300 bar. However, using a ratio as low as of 58 g CO2 /g, 78% of the initial caffeine was recovered under the same conditions. The remaining caffeine seemed to be not removable under the conditions studied, due to the strong interactions existing between caffeine and the plant matrix and the important limitations to internal mass transfer. Caffeine was not extracted pure, but in a mixture of fats and small quantities of pigments, which could be easily separated by simple water washing and subsequent evaporation, resulting in high purity extracts. Thus, coffee husks are a very interesting option as a raw material from an economic and industrial point of view since they are an abundant residue of the coffee industry with a high caffeine content, most of which is simply extracted. The low cost of the raw material and the employment of moderate CO2 ratios would result in low production costs. However, an exhaustive economical analysis should be made to establish if the combination of this benet together with the selling of the caffeine and the husks could counterbalance the relatively high installation costs. Acknowledgement The authors thank Natraceutical Group for the nancial support. References
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