Separation and Purication Technology 48 (2006) 94105

The effects of both preparation method and season on the supercritical extraction of ginger
S. Balachandran a , S.E. Kentish a, , R. Mawson b
a

Co-operative Research Centre for Bioproducts, Department of Chemical and Biomolecular Engineering, University of Melbourne, Vic. 3010, Australia b Food Science Australia, Sneydes Road, Werribee, Vic. 3030, Australia

Abstract Experiments were conducted to study and compare the supercritical extraction of oleoresin from fresh, oven-dried and freeze-dried ginger samples. When new season water-rich fresh ginger rhizomes were utilised, higher extraction yields resulted relative to both the oven-dried and freeze-dried samples. Mathematical modelling of experimental data conrmed that this yield enhancement resulted principally from an enhancement in the effective diffusivity within the ginger particle. However, when a more brous and less moist old season ginger supply was utilised, the mass transfer rate in all samples declined. Further, the reduction in effective diffusivity was most pronounced for the fresh ginger sample. HPLC analysis showed that the ginger extract obtained from the oven-dried feed contained proportionately less gingerols and more shogaols due to thermal degradation. This conrms that the utilisation of fresh ginger rhizomes for supercritical extraction would lead to a more pungent and naturally avoured extract. However, there are a number of signicant disadvantages to this approach from a manufacturing perspective that would need to be considered. 2005 Elsevier B.V. All rights reserved.
Keywords: Supercritical extraction; Mass transfer; Moisture level; Ginger; Shrinking core model

1. Introduction The extraction of soluble essences from natural products using supercritical uids has signicant advantages over conventional methods such as steam distillation and Soxhlet extraction [13]. Higher solubilities, mass transfer rates and selectivities make supercritical uid extraction (SFE) more attractive. The selectivity of the compound to be extracted is dependent on the density of the supercritical uid, which can be altered by varying process conditions. The low latent heat of evaporation and high volatility of the solvent makes the extract free from residual solvent. Moreover, extraction can be conducted at low temperatures, which helps preserve thermally degradable food products [46]. Supercritical carbon dioxide (Sc. CO2 ) is considered to be a particularly useful solvent in the extraction of essential oils from vegetable substrates due to its environmentally

Corresponding author. Tel. +61 3 8344 6682; fax: +61 3 8344 4153. E-mail address: sandraek@unimelb.edu.au (S.E. Kentish).

benign nature. However, one limitation to the use of this uid for extraction is its non-polar nature. The use of co-solvents and modiers has been studied by various researchers [79] to overcome this limitation. Water and solvents such as ethanol, isopropanol, acetone and methanol are among the more acceptable co-solvents for food grade products. The advantage of water over organic solvents is that the extraction process remains clean and non-toxic. Organic solvents are highly ammable, which further restricts their use in industry. A number of research groups have also considered the inuence of the substrate moisture level on extraction yield using Sc. CO2 . Hubert and Vitzthum [10] observed that the normal moisture content of tobacco had to be increased by up to 25% to affect nicotine extraction. Zosel [11] reported that pre-soaking coffee beans aids the removal of caffeine. Similar pre-soaking characteristics are reported by Nguyen et al. [12] to increase the extraction of vanilla from vanilla beans. Leeke et al. [13] studied the degree of extraction of essential oil from a model herb (Origanum vulgare L. ssp. Virens letswarrt)

1383-5866/$ see front matter 2005 Elsevier B.V. All rights reserved. doi:10.1016/j.seppur.2005.07.008

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Nomenclature ap Bi C Ci Csat De Ds Kf m q q q0 r rc R t u VCO2 Vg X Xi Y specic surface area of the particle (m2 /m3 ) Biot number, Kf R/De solute concentration in the bulk uid phase (kg/m3 ) solute concentration in the pore volume of the solid (kg/m3 ) saturation concentration of the solute in the CO2 phase (kg/m3 ) effective solid diffusivity (m2 /s) solid diffusivity (m2 /s) external mass transfer coefcient (m/s) partition coefcient (m3 of ginger/m3 of CO2 ) solid phase solute concentration (kg/m3 ) average solid phase solute concentration (kg/m3 ) initial solid phase solute concentration (kg/m3 ) radial co-ordinate in particle (m) radius of the core (m) radius of the solid particle (m) time (s) supercial velocity (m/s) volume of carbon dioxide in the vessel (m3 ) volume of ginger in the vessel (m3 ) dimensionless concentration in the bulk uid phase, C/Csat dimensionless concentration within the pore volume, Ci /Csat dimensionless concentration in the solid phase, q/qo

Greek letters dimensionless time, De /R2 t dimensionless radial coordinate of the particle, r/R c dimensionless core radius in the particle, rc /R

with varying bed moisture levels. They observed that extraction was enhanced due to changes in the transport properties of the solvent and a decrease in intraparticle resistance of the solute inside the particles. Fahmy et al. [14] studied the effect of modiers (water, methanol and acetonitrile) in the extraction of solutes from clay, soil and plant matrices. They showed that the swelling properties of water can also enhance the extraction process. Conversely, an inhibiting effect of water on extraction has also been reported. Antunes et al. [15] observed an increase in extraction of organochlorines from freeze-dried sh samples compared to fresh sh llet samples, but could not explain the inhibiting effect of water. Smyth et al. [16] observed a decrease in extraction of naphthalene from a soil matrix, when the moisture content of the matrix was increased from 10 to 20%. A condition

termed water bridging which lowers the interfacial areas and increases path length was considered to be controlling the mass transfer process. Recently, Yoda et al. [17] reported a lower yield from Stevia rebaudiana containing 510% moisture at supercritical conditions, but obtained higher yield at subcritical conditions. While most of the literature has shown that a high moisture content has a positive effect on extraction processes, the focus has tended to be on analytical aspects such as the amount of moisture required and the composition of the resultant extracts. Information on the effects of moisture content on the intrinsic kinetic parameters is lacking. In this work, the inuence of water on the extraction kinetics, solubility parameters and overall yield of the soluble essence from a typical herb is studied. The model herb used was Ginger (Zingiber Ofcinale Roscoe), which belongs to the Zingiberceae family. Ginger is a food crop used worldwide as a spice and food-avouring agent. Its neutraceutical properties have been of interest in food processing and in the pharmaceutical industries [18]. The essential oil consists of monoterpenes and sesquiterpenes, which contribute to the characteristic avour of ginger. The more volatile oleoresin is responsible for the pungent avour of ginger and is a source of anti-oxidants. The main pungent compounds are the series of homologues called gingerols, which are prominent in fresh ginger. 6-Gingerol(5-hydroxyl-1-(4hydroxy-3-methoxyphenyl)decan-3-one) is the most abundant constituent in the gingerol series [19]. Most works in the scientic literature consider the supercritical extraction of oleoresin from dried ginger. Spiro and Kandiah [20] studied the extraction kinetics of sun-dried Jamaican ginger at various temperatures and pressures. They reported the rate constant of the extraction process as a function of three stages: a short washing stage, a longer fast extraction stage and a slow extraction stage. They note that water appeared to play a role in the extraction kinetics, but conclude that the increase in solvent density would be dominating rather than the entraining effect of water. Roy et al. [21] propose a shrinking core model to explain the extraction of ginger oil from freeze-dried ginger. They showed that solvent density and solute vapour pressure dominate the mass transfer rate. Yonei et al. [22] studied the inuence of various process parameters on the extraction rate of Chinese type ginger. Martinez et al. [23] solved the differential mass balance model with logistic equations to explain their results. While these workers consider dried ginger, it is well known that during the industrial drying process, gingerol dehydrates to a series of homologues called shogaols [19,24], which have less pungent properties and lower antioxidant activity. Only Bartely et al. [25,26] have considered the extraction of the more pungent fresh ginger. While they did not collect any kinetic data, they identied the chemical composition of the extract using gas chromatography (GC)mass spectrometry (MS) and GCelectrospray mass spectrometry (EMS). They show that as a result of the drying process, approximately 50% of the gingerols convert into shogaols

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and the remainder to zingerone and hexanal. They argue that the compositional changes that occur during drying would be perceived by the consumer as a decrease in the pungency and that supercritical uid extraction could produce a avour extract that preserved the natural characteristics of the fresh product. The main objective of this work is thus to further investigate the effect of the moisture content and drying method on the supercritical extraction of a pungent oleoresin from ginger. We aim to determine whether the use of a fresh ginger substrate could improve both the quality and quantity of the extract and the efcacy with which it could be produced.

batches were freeze dried using a commercial scale freeze drier which was operated for 4872 h at 20 C and high vacuum. Further samples from both batches were oven dried in a laboratory oven for 6 h at 60 C, to replicate the industrial drying conditions commonly used in Australia. It was assumed that the freeze drying process produced a product of negligible water content. Based on the total weight loss during freeze drying, the initial moisture content of the original sample was then calculated as 96 wt% for Batch A and 83.5 wt% for Batch B. Similarly, the residual water content of the oven dried samples was calculated to be <1 wt% for both batches. 2.2. Experimental method

2. Experimental 2.1. Materials and sample preparation Two different batches of fresh ginger rhizomes were bought from Queensland through a local market during February of 2003 and 2005. While these batches were purchased in the same calendar month, Batch A was a new season product with a white-yellow esh appearance. Conversely, Batch B was old season ginger from the previous harvest and was characterised by a more brous and darker appearance. Both batches were refrigerated at 4 C to avoid loss of freshness. Each rhizome was peeled and cut into uniform size cylinders using an industrial cutting machine. The cylinder height was adjusted to ensure the total volume was equivalent to that of a sphere of identical diameter. Samples from both A 150 ml stainless steel batch extractor was used in all experiments. This vessel is immersed in a temperature controlled water bath. As shown in Fig. 1, the extraction vessel is coupled with an online UVvis spectrophotometer with independent temperature control for kinetic extraction studies. A magnetic gear drive pump (model 1805/IEC71, Micropump) provides circulation through a stainless steel high-pressure cell with sapphire windows (Graseby Specac, UK) which was placed within the spectrophotometer. By adjusting the valve positions, the ow can be directed through a baseline loop that bypasses the sample vessel, or through a sampling loop that includes the extractor. At the beginning of each experiment, the extraction vessel was packed with 3 mm glass beads and the required amount of ginger. The unit was then heated to the desired temperature.

Fig. 1. Process ow sheet for Sc. CO2 extraction experiments.

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Liquid carbon dioxide was delivered to a syringe pump (ISCO 260D, USA) and pressurised to the desired pressure. The baseline loop was then pressurised to the desired operating pressure. Baseline spectral measurements for pure CO2 were recorded in the spectrophotometer. Experiments only proceeded if this baseline reading was within 0.05 of zero. The extraction vessel was then pressurised with CO2 . This pressurization takes approximately 810 min and spectral data collection commenced only at the end of this stage. The spectral data was recorded at 2 min intervals by the spectrophotometer as extraction proceeded. Pressure, temperature and ow rates were checked periodically to ensure consistency. After completion of the extraction process, CO2 was depressurised through a needle valve to a Dreschel bottle containing 30 ml of methanol. This methanol sample was ltered and used for HPLC analysis. The CO2 was vented to atmosphere through a rotameter (MFG Co. Ltd., UK) and a gas ow meter (Alexander Wright and Co. Ltd., UK), which was used to verify the amount of CO2 used in the experiment. The unit was cleaned with pure CO2 and organic solvents. Finally, a baseline scan was performed to conrm that no solute remained. Experiments were conducted at 160 bar pressure and 40 C temperature with three different particle sizes of fresh, ovendried and freeze-dried ginger samples. Previous work has indicated that these conditions are the most suitable for ginger extraction [26]. Higher temperature can cause gelatinisation of starch, which may block the cell walls and disturb mass transport of the solute [27]. Most researchers [18,19,28] have considered the pungent compounds, which are principally gingerols and shogaols, to have a spectral absorbance peak at 280 nm. This assumption is used in this work to record kinetic data. The solvent concentration was calculated using BeerLamberts law [29] from the absorbance recorded at 280 nm in the spectrophotometer, using an extinction coefcient of 3000 l/(mol cm) [28]. In preliminary experiments, the quantity of ginger required to give acceptable absorbance readings was estimated. It was found that for Batch A, much less ginger was
Table 1 Physical property data used as input to the models Pressure (bar) Temperature (K) CO2 density (kg/m3 ) CO2 viscosity (kg/(m s)) CO2 -gingerol diffusivity in supercritical phase (m2 /s) External mass transfer coefcient Kf (m/s) Supercial velocity in bed (m/s) Batch A ginger Fresh Bulk density (kg/m3 ) Sample mass (g) Sample dry mass (g) Initial gingerol content (wt% of dry solids) 1060 9 0.45 2.4 Freeze-dried 74 1 1 2.4

required (on a dry solids basis) for the fresh ginger than for the oven-dried ginger and the freeze-dried sample required an intermediate quantity. The exact quantities used are given in Table 1. Some runs are repeated in duplicate to conrm the accuracy of the experimental method. These duplicate runs suggested that concentration data could be replicated to within 10% deviation. 2.3. Chromatographic analysis High-performance liquid chromatography (HPLC), gas chromatographymass spectrometry (GCMS) and thin layer chromatography (TLC) have all been used in the past to analyse supercritical ginger extract [30,31]. The pungent compounds can be separated as groups by TLC on silica gel plates, but individual components cannot be well resolved. With GCMS, pungent compounds can undergo spontaneous thermal degradation and dehydration. Connell [19] has concluded that GC is an unsatisfactory tool to examine gingerol and shogaol levels. Conversely, other researchers [26,32] have successfully used GCMS to analyse the concentration of the essential oil components. HPLC is considered to be the best analytical tool by many researchers [30,33] to quantify particular compounds of similar absorbance that are present in low concentrations within an extract. In the present work, a Shimadzu liquid chromatography system equipped with a model LC-10 AT VP solvent delivery pump, a Rheodyne 7125 injector (loop volume of 20 l) and a SPD-10AV VP variable wavelength UVvis detector was used. A reversed phase column (Exil ODS RP-18, 25 cm 4.6 mm, 5 m) was purchased from SGE. Analysis was conducted using an isocratic method with a mixture of HPLC grade methanol and water (70:30, v/v). The analysis time was 30 min. The mobile phase ow was 1 ml/min and detection was based on UV absorption at 280 nm. The compounds were identied and quantied based on retention times using both published data from the literature and a 6-gingerol HPLC standard purchased from Wako Pure Chemical Industries, Osaka, Japan (99% purity).

160 313 796.4 0.06869 103 7.2 109 6 105 1.9 103 Batch B ginger Oven-dried 578 1.74 1.71 Fresh 1077 9 1.5 2.2 Freeze-dried 196 1.5 1.5 2.2 Oven-dried 714 1.74 1.71

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3. Mathematical model development is The basic assumptions made in developing a mathematical model replicate those used by other researchers. That is: (a) The extraction vessel is considered to be lled with homogeneous spherical particles of uniform size. (b) Several compounds will be simultaneously extracted from the ginger. However, the model considers the behaviour only of a single pseudo-component, characterized by 6-gingerol with a spectrophotometric absorbance peak at 280 nm. A similar assumption has been made by a number of workers [18,19,28]. (c) There is a continuous low ow through the extractor because of the ow loop through the spectrophotometer. While this ow is assumed to enhance the external mass transfer coefcient through standard correlations, the extractor is otherwise considered as a well-mixed reactor with minimal axial and radial concentration gradients and negligible pressure drop. (d) The system is isothermal and isobaric. (e) The physical properties of Sc. CO2 are constant. Further development of mathematical models requires consideration of the specic microscopic structure of the solid. In the case of the fresh ginger particle, the solute is evenly dispersed through a water-lled matrix. During extraction, this solute will migrate through this water-rich phase to the particle surface where it will desorb into the supercritical uid phase. Conversely, in a dried particle, the supercritical uid must rst diffuse into the particle pore structure. As the extraction vessel is not evacuated prior to experiments, any residual air content within the pores may retard this solvent penetration. Desorption of solute into the solvent will then occur within the particle pores, followed by migration through Sc. CO2 uid phase to the particle surface and beyond to the bulk phase. 3.1. Solid diffusion model for fresh ginger For fresh ginger, we assume that the gingerol solute is already contained within the water-rich matrix. The rate determining steps are diffusion to the particle surface, characterised by a diffusion coefcient (Ds ) and external mass transfer, characterised by a uid phase mass transfer coefcient (Kf ). Desorption from the water-rich phase into the supercritical phase is assumed not to be rate controlling but characterised by an equilibrium relationship: C(R) = mq(R) (1)

The equation for the diffusion within the spherical particle 2 q 2q q = Ds + t r 2 rr

(3)

The initial and the boundary conditions for Eqs. (1)(3) are C[t = 0] = 0 (0 r R) q[r, 0] = q0 q =0 (r = 0; t 0) r q = Kf [C C(R)] (r = R; t 0) Ds r These equations were converted into a dimensionless format and then solved using a nite difference approach with the Mathematica 4.1 software package. 3.2. Shrinking core model for freeze-dried ginger For freeze-dried ginger we use the shrinking core model developed by Goto et al. [34], but adapted to a batch extraction process. With this model, an inner non-extracted core shrinks as extraction progresses. Solute is desorbed from the boundary of the non-extracted core, resulting in a sharp dissolution front with a saturated uid concentration (Csat ) at this point. The solute then diffuses through the pore structure with an effective diffusivity (De ). This is followed by mass transfer through the bulk uid phase with a mass transfer coefcient (Kf ). The rate of solute mass transfer into the bulk uid phase can be expressed in terms of the gas phase mass transfer coefcient (Kf ) Vg dC = ap Kf [Ci (R) C] dt VCO2 (4)

Eqs. (2) and (4) are similar, except that the mass transfer is from the solid bulk in the rst case and from the pore volume in the latter case. Similarly, the rate of solute mass transfer from the solid phase can also be written in terms of the gas phase mass transfer coefcient (Kf ): d q = ap Kf [C Ci (R)] dt Diffusion in the pore is given by: De r2 r r2 Ci r =0 (6) (5)

The differential mass balance in the bulk uid phase surrounding each particle is Vg ap Kf [C(R) C] dC = dt VCO2 (2)

The average solid phase concentration ( q) is described as a function of particle diameter: q rc = qo R

3

(7)

In Eq. (2), ap = 3/R is the specic surface area of the particle.

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The initial and boundary conditions are given by: C[t = 0] = 0 (0 r R) q[r, 0] = q0 Ci [r, t ] = Csat (r = rc ; t 0) Ci De = Kf [C Ci (R)] (r = R; t 0) r The above equations and the initial and boundary conditions can be expressed in terms of the following dimensionless variables as: 3BiVg 1X dX = d VCO2 1 Bi(1 1/c )
3 Y = c

(8) (9)

The respective initial and boundary conditions are: X[ = 0] = 0 (0 c 1) Y [c , = 0] = 1 X = Bi[bX Xi (1)] (c = 1; 0) c (0 c 1; = 0) Xi = 1 The above dimensionless differential equations and their respective initial and boundary conditions were solved numerically using a fourth order RungeKutta method within the Mathematica 4.1 software package. While the freeze-dried ginger was modelled using this approach, we elected not to model the oven-dried results. This data tended to be less reproducible than that obtained for fresh and freeze-dried samples. We believe this increased variability arose from the less uniform particle shapes and size distributions that resulted from the oven drying process. 3.3. Estimation of properties for theoretical analysis In order to utilise the above models, the viscosity of Sc. CO2 was estimated using the empirical correlation of Jossi et al. [35]. The binary diffusion coefcient for gingerolcarbon dioxide at supercritical conditions was calculated by the method proposed by Takahashi [36]. The estimated values are shown in Table 1. As a major portion of the extractor bed was dominated by glass beads, it is best to calculate the bed void fraction based on the packing of these beads. A value of 0.5 was calculated from the ratio of the volume occupied by glass beads to that of the total volume of bed. This bed void fraction was used to calculate the interstitial velocity of solution through the bed. The external mass transfer coefcient Kf , was estimated from the empirical correlation reported by Wakao et al. [37] for solids and liquids or gases at low pressure regions. The correlation is valid for 3 < Re < 3000, which was within the range of our experimental conditions. The bulk density of individual ginger particles is calculated from the ratio of mass to volume.

To determine the initial concentration of gingerol compounds in each batch, about 0.1 g of ground freeze-dried ginger was mixed with 30 ml of methanol in a conical ask and stirred using a shaker table. Samples were taken every 24 h and the spectrophotometric absorbance recorded at 280 nm. Sampling continued until this absorbance reached a constant value. Generally, extraction was complete in 24 h. On this basis, the initial amount of gingerols/pungent compounds was calculated as 2.4 wt% (on a dry solids basis) for the new season Batch A ginger and 2.2 wt% for the old season Batch B ginger. These values are consistent with literature values [18,24] that vary from 1.9 to 3 wt%. It is normally found that the gingerol in old season ginger is higher than that in the new season product. While on a dry ginger basis this is not reected in the present results, on a total solids basis, the gingerol is signicantly higher in Batch B, because of the lower water content of this material. On a total solids basis, the respective gingerol contents in the fresh ginger of Batch A and Batch B are qo = 1.3 kg/m3 and 3.9 kg/m3 , respectively. Further, the Batch B ginger had probably been stored for a number of months, and such storage is also known to lead to a reduction in gingerol content [24]. It was difcult to use this approach to determine the quantity of gingerol in the fresh ginger samples. However, Zhang et al. [24] have shown that contrary to oven drying, freeze-drying does not result in a signicant loss of gingerol. Consequently, we utilised the freeze-dried values in our fresh ginger modelling.

4. Results and discussion Figs. 2 and 3 show the concentration of pungent compounds (the pseudo-component gingerol) as a function of both time and particle size for fresh and freeze-dried ginger samples from Batch A. Results with all ginger samples showed a signicant dependence upon particle size which is consistent with the work of other researchers [21,22]. Of greater interest in the present work is that the water abundant fresh ginger from Batch A shows a signicant

Fig. 2. Concentration of pungent compounds in the bulk uid phase for different particle sizes of Batch A fresh ginger. Continuous curves from model. Amount of ginger used (dry basis) = 0.45 g.

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Fig. 3. Concentration of pungent compounds in the bulk uid phase for different particle sizes of Batch A freeze-dried ginger. Continuous curves from model. Amount of ginger used (dry basis) for 4.02 mm particle size = 1.5 g, and for 5.8 and 7.7 mm particle size = 1 g.

Fig. 5. Yield of pungent compounds vs. extraction time for Batch B, based on the on-line spectrophotometer results.

increase in the extraction of oleoresin over that of the dried ginger. In these experiments, it was observed that extraction was higher from fresh ginger even though the extractor was loaded with 23 times less solids on a dry weight basis. Fig. 4 shows the extraction yield (i.e. the grams of pungent compounds extracted divided by the grams of dry solids in the feed) as a function of time for fresh, freeze-dried and oven-dried ginger of around 6 mm particle size. This relative increase can be related to three possible factors: (a) the dissolution of water into the supercritical phase enhancing the solubility of gingerol through an entrainer type mechanism. Under the experimental conditions, 0.074 g of water would be expected to dissolve in the working volume of Sc. CO2 (110 ml) at equilibrium [38]. This is less than 1% of the total water present in the fresh ginger feed; (b) the water-swollen ginger providing less mass transfer resistance to the intra-particular movement of gingerol to the particle surface; (c) a signicant loss of gingerol from the oven dried feed as a consequence of the drying process itself.

To conrm whether these results persisted across a range of ginger rhizome conditions, and in order to obtain HPLC results, the above experiments were repeated with Batch B. However, as shown in Fig. 5, in this case, the results are reversed with freeze-dried ginger providing a higher yield than the fresh product. All yields are also signicantly lower with Batch B. 4.1. Modelling results The best t to the fresh ginger data was obtained by varying the effective diffusivity (Ds ) and partition coefcient (m) in the solid diffusion model until the sum of squared errors was minimised. In dimensionless format, the data for different particle sizes collapse onto a single curve, making this optimisation relatively straightforward. Fig. 2 shows the best t curves obtained while Table 2 summarises the optimised Ds and m values and the standard errors obtained for both batches. The predicted effective diffusivity for Batch A is two orders of magnitude higher than that of Batch B ginger. This indicates that the resistance to solute diffusivity is signicantly higher in Batch B, which would lower the yield at any given time relative to Batch A experiments. Further, it appears that the optimised partition coefcient is also lowered for Batch B experiments. This decrease in the partition coefcient may reect the lower water level in this batch. As there is less water available, and the mass transfer rate of water into the supercritical phase will also decline, this water may not be as available to act as an entrainer and enhance the equilibrium concentration of gingerol. However, the change in the partition coefcient may also indicate that the use of a linear equilibrium relationship is too simplistic. As shown in Table 4, once the gingerol concentrations in the solid phase are taken into account, the model predicts comparable saturation concentrations in the supercritical phase. In modelling the freeze-dried ginger the unknown parameters are the effective diffusivity (De ) and saturation concentration value (Csat ). The best t was again obtained by varying these parameters until the sum of squared errors was

Fig. 4. Yield of pungent compounds vs. extraction time for Batch A, based on the on-line spectrophotometer results.

S. Balachandran et al. / Separation and Purication Technology 48 (2006) 94105 Table 2 Predicted effective diffusivity and partition coefcient values for different batches of fresh ginger Fresh ginger Batch A Partition coefcient m of Effective Diffusivity Ds (m2 /s) Particle size (mm) (m3 ginger/m3 of CO2 ) 4.7 0.0023 2.5 1.6 1011 6 0.0016 Batch B

101

7.9 0.0010

1.0 4.8 1013 6.5 0.0011

Standard error in concentration (kg/m3 )

Table 3 Predicted effective diffusivity and saturation concentration value for different batches of freeze-dried ginger Freeze-dried ginger Batch A Saturation concentration Csat Effective rst stage Diffusivity De (m2 /s) second stage Particle size (mm) Standard error in concentration (kg/m3 ) 3.0 1.4 1012 9.5 1013 5.7 9.3 104 Batch B 5.0 9.2 1013 4.4 1013 6.3 1 103

4.02 3.9 103

7.5 6.7 104

(kg/m3 )

minimised. Fig. 3 shows the best t curves obtained while Table 3 summarises the optimised De and Csat values and the standard errors obtained. While it was possible to t the experimental data for freeze-dried ginger with a single diffusivity value, a much better data t could be obtained by using one value of De for the initial period of the experiments and a lower value in latter stages. This slowing in mass transfer rate has been discussed by Sovova et al. [39] while modelling the extraction of vegetable oil. They consider particles to have two different regions; one region which is broken during sample preparation and the other which has intact cells. Marrone et al. [40] also considered a similar approach to model the extraction of almond seeds. They considered two different solute concentrations that have different mass transfer resistances inside the solid particles. This hypothesis was conrmed by performing scanning electron microscopy of the particles. In this work, the model shows that the transition of effective diffusivity occurred at a constant penetration depth inside all particles. This conrms that the solute is initially extracted from ruptured cells and is unhindered by the cell membranes and walls. In the later stages, extraction occurs from intact cells and hence the diffusional resistance increases. The predicted effective diffusivity for Batch A is again larger than for Batch B ginger but the difference is less dramatic than in the fresh ginger case. Further, the optimised saturation concentration Csat is in this case higher for Batch B than Batch A ginger.

versely, in the freeze-dried ginger model, these pungent components partition into the supercritical phase, internally at the core radius rc . The diffusivity therefore represents diffusion through a supercritical phase. To compare these two diffusivities directly, it is necessary to convert the fresh ginger diffusivity to a parameter based on the supercritical phase. As discussed by Do and Rice [41] the appropriate conversion is: De = Ds m (10)

Similarly, in the freeze-dried modelling, the partition coefcient is not explicitly calculated, rather only the equilibrium saturation concentration, Csat . Again, for comparison purposes, this can be compared to the saturation concentration that would be in equilibrium with fresh ginger, i.e. Csat = mqo (11)

Table 4 summarises the parameters for both fresh and freeze-dried ginger on this basis. The predicted effective diffusivity for the fresh ginger from the water-rich new season Batch A is ve times higher than the value for the freezedried ginger samples. This high diffusivity may be explained on the basis of the structure of the matrix. Batch A contains
Table 4 Comparison of kinetic parameters of fresh and freeze-dried ginger of Batch A and Batch B Kinetic parameters Batch A 6.4 1012 3.2 1.2 1012 3.0 Batch B 4.8 1013 3.9 6.8 1013 5.0

4.2. Comparison of kinetic parameters The diffusivity calculated from the fresh ginger simulations represents the ow of pungent compounds through the water-saturated solid phase, with the partition into the supercritical phase occurring at the outer solid surface. Con-

Fresh ginger Freeze-dried ginger

De (m2 /s) Csat (kg/m3 ) De (m2 /s) Csat (kg/m3 )

The effective diffusivity (De ) and saturation concentration (Csat ) values for the fresh ginger are estimated from Eqs. (10) and (11).

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Table 5 Comparison of predicted effective diffusivity (De ) value and saturation concentration (Csat ) values with literature values Reference works (process conditions) Roy et al. [21]: extraction with freeze dried ginger (245 bar, 40 C) Goto et al. [34]: extraction studies with rapeseed (205 bar, 51 C) Goodarznia and Eikani [42]: extraction with rapeseed, basil, marjoram (100 bar, 40 C) This work: fresh ginger (160 bar, 40 C) This work: freeze-dried ginger (160 bar, 40 C)
a

Literature value De (m2 /s) 2.5 1010 1.5 1010 4.7 1013 Csat (kg/m3 ) 1.25 2.5

Adjusted value to 160 bar, 40 Ca De (m2 /s) 4.2 1010 1.7 1010 2.6 1013 6.4 1012 1.4 1012 Csat (kg/m3 ) 0.65 2.5 3.2 3.0

De values are adjusted using the method of Takahashi [36]. Csat values are adjusted using the empirical model suggested by Chrastll [44].

96% water, so there is low tortuosity and limited obstruction to the ow path. This is in good agreement with the mechanism discussed by Leeke et al. [13] who consider the higher solute diffusivity and the swelling of the matrix as reasons for the enhancement of extraction yield from water lled matrices. When the more brous old season ginger is used in place of Batch A, the diffusivity for both fresh and freeze-dried ginger falls. However, the decline for the fresh ginger product is larger, leading to the reversal in the relative yields from these products. This is consistent with the results of other workers who nd that high water contents are necessary to enhance mass transfer rates. The saturation concentrations in the four cases modelled are comparable, indicating that the entraining effect of water is marginal. The saturation concentration for Batch B is slightly larger and this may be the result of changes in the pungent compound composition between batches. Changes in the relative proportions of the gingerols and shogaols that absorb at 280 nm will affect the partition coefcient for the overall pseudo-component modelled in this work. The predicted kinetic parameters have been compared to literature values and are summarised in Table 5. Predicted effective diffusivity values are two orders of magnitude lower than the values predicted by Roy et al. [21] and Goto et al. [34] but an order of magnitude higher than the values predicted by Goodarznia and Eikani [42] and Reverchon et al. [43]. Reverchon et al. [43] and Roy et al. [21] have pointed out that such differences can be due to different mass transfer mechanisms, cell structure, mechanisms of solute extraction and molecular sizes and hydrophilic properties of the solute.

We further predict a signicantly higher saturation concentration of gingerol in the supercritical phase for the freezedried ginger model relative to the values provided by Roy et al. [21]. This may again be caused by sample variation. Further, the results obtained by Roy et al. [21] were at a signicantly higher CO2 pressure and this will affect the composition of the extract. Both Roy et al. [21] and the present work measure gingerol concentration using only the absorbance at 280 nm. This is a crude concentration measurement and changes in the composition of the pungent extractives will inuence the concentration equilibria. 4.3. High-performance liquid chromatography In order to validate the concentration data, HPLC analysis was also completed on the supercritical extracts from Batch B. As indicated in the experimental procedure, the Sc. CO2 is exhausted into a methanol solution after extraction. However, we found that much of the nal extract also deposited in the tubing between the depressurising valve and the methanol collection vessel. We consequently collected this material by washing the tubing with a further quantity of methanol. Both samples were analysed using the UV spectrophotometer in order to quantify the total recovery of oleoresin before analysis with HPLC. As shown in Table 6, these spectrophotometer results indicated that we had recovered between 60 and 75% of the extract based on the nal on-line spectrophotometric results. A number of compounds are eluted during HPLC analysis. Fig. 6 shows the different compounds eluted from fresh,

Table 6 Concentration of 6-gingerol in different samples of Batch B ginger analysed by a HPLC technique Ginger type Spectrophotometeric analysis of total pseudo-component Yield (wt% of dry solids) Fresh ginger Oven-dried Ginger Freeze-dried Ginger Tube wash Separator Tube wash Separator Tube wash Separator 0.070 0.051 0.11 0.048 0.16 0.028 Total yield (wt% of dry solids) 0.12 0.16 0.19 HPLC analysis of 6-gingerol Yield (wt% of dry solids) 0.013 0.012 0.025 0.0011 0.047 0.0026 Total yield (wt% of dry solids) 0.025 0.026 0.05 20 16 26 Percentage of 6-gingerol in pseudo component

Tube wash: solvent from washing the tubings with methanol; separator: solvent from Dreschel bottle.

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Fig. 6. HPLC chromatograms of: (A) fresh ginger extract; (B) oven-dried ginger extract; and (C) freeze-dried ginger extract. Peaks are: (a) 6-gingerol; (b) 8-gingerol; (c) 6-shogaol; (d) 8-shogaol; and (e) not identied.

oven-dried and freeze-dried ginger extract, which are identied based on retention times, in comparison with literature results [30,33]. A qualitative analysis of these chromatograms shows that the fresh ginger extract has a higher proportion of gingerols compared to both the oven-dried and freeze-dried ginger extracts, which are richer in shagoals. Quantication of all these compounds is not possible as 6-gingerol is the only standard available commercially. However, as can be seen from Table 6, 6-gingerol indeed only represents around 20% of the total quantity of compounds that absorb at this wavelength. The proportion of 6-gingerol in the oven-dried sample is lower than that for both the freezedried and fresh samples. This is probably due to thermal degradation of 6-gingerol to shogaols during the oven drying process, as also observed by Bartley et al. [26] and Zhang et al. [24]. 4.4. Other issues While the results presented above show signicant positive benets from the use of a fresh ginger feed, there are some disadvantages to this approach that must also be con-

sidered. Firstly, while the yield may be higher on a dry solids basis, the volume of fresh ginger relative to the oven-dried product is also higher. Given the high capital cost associated with supercritical extraction vessels, the volume of ginger that can be accommodated in a given run must also be considered. Thus, on a volume basis, around 10 times as much oven or air-dried ginger could be accommodated within such a vessel relative to the fresh product. Given this higher load, there would indeed be a greater yield per run from the dried product than from the fresh product. Secondly, it is easier to store and transport a dried product relative to a fresh one. If fresh ginger was to be utilised the extraction facility would need to be located close to the harvesting site. It may also be necessary to add preservatives to any fresh product being stored for long periods and this would negate the fresh product marketing advantage. Finally, this work was based on relatively large particle sizes in order to extract kinetic data. In commercial practice much smaller particle sizes are utilised. The yield enhancement described above for fresh ginger would increase as the particle size is reduced. However, forming discrete particles of very small size from fresh ginger would be difcult and

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S. Balachandran et al. / Separation and Purication Technology 48 (2006) 94105 [6] M.A. McHugh, V.J. Krukonis, Supercritical Fluid Extraction: Principles and Practice, 1, second ed., ButterworthHeinemann, Boston, USA, 1994. [7] S.S.T. Ting, S.J. Macnaughton, D.I. Tomasko, N.R. Foster, Solubility of Naproxen in supercritical carbon dioxide with and without cosolvents, Ind. Eng. Chem. Res. 32 (1993) 14711481. [8] T. Wang, H. Wang, Solubility of diethylammonium diethyldithiocarbamate in supercritical carbon dioxide with ethanol as the co-solvent, Chem. Eng. Process. 42 (2002) 6165. [9] J.M. Dobbs, J.M. Wong, R.J. Lahiere, K.P. Johnston, Modication of supercritical uid phase behaviour using polar solvents, Ind. Eng. Chem. Res. 26 (1987) 5665. [10] P. Hubert, O.G. Vitzthum, Fluid extraction of hops, spices and tobacco with supercritical gases, Angew. Chem. Int. Ed. Engl. 17 (1978) 710715. [11] K. Zosel, Separation with supercritical gases: practical applications, Angew. Chem. Int. Ed. Engl. 17 (1978) 702709. [12] K. Nguyen, P. Barton, J.S. Spencer, Supercritical carbon dioxide extraction of vanilla, J. Supercrit. Fluids 4 (1) (1991) 4046. [13] G. Leeke, F. Gaspar, R. Santos, Inuence of water on the extraction of essential oils from a model herb using supercritical carbon dioxide, Ind. Eng. Chem. Res. 41 (2002) 20332039. [14] T.M. Fahmy, M.E. Paulaitis, D.M. Johnson, M.E.P. Mcnally, Modier effects in the supercritical uid extraction of solutes from clay, soil and plant materials, Anal. Chem. 65 (1993) 1462 1469. [15] P. Antunes, O. Gil, M.G. Bernardo-Gil, Supercritical uid extraction of organochlorines from sh muscle with different sample preparation, J. Supercrit. Fluids 25 (2) (2003) 135142. [16] T.J. Smyth, R.G. Zytner, W.H. Stiver, Inuence of water on the supercritical uid extraction of naphthalene from soil, J. Hazard. Mater. 67 (2) (1999) 183196. [17] S.K. Yoda, M.O.M. Marques, A.J. Petenate, M.A.A. Meireles, Supercritical uid extraction from Stevia rebaudiana Bertoni using CO2 and CO2 + water: extraction kinetics and identication of extracted components, J. Food Eng. 57 (2003) 125134. [18] V.S. Govindarajan, Ginger chemistry, Crit. Rev. Food Sci. Nutr. 17 (1) (1982) 196. [19] D.W. Connell, The pungent principles of ginger and their importance of ginger product, Food Technol. Aust. November (1969) 570 575. [20] M. Spiro, M. Kandiah, Extraction of ginger rhizome: kinetic studies with supercritical carbon dioxide, Int. J. Food Sci. Technol. 25 (1990) 328338. [21] B.C. Roy, M. Goto, T. Hirose, Extraction of ginger oil with supercritical carbon dioxide: experiments and modelling, Ind. Eng. Chem. Res. 35 (1996) 607612. [22] Y. Yonei, H. Ohinata, R. Yoshida, Y. Shimizu, C. Yokoyama, Extraction of ginger avour with liquid or supercritical carbon dioxide, J. Supercrit. Fluids 8 (2) (1995) 156161. [23] J. Martinez, A.R. Monteiro, P.T.V. Rosa, M.O.M. Marques, M.A.A. Meireles, Multicomponent model to describe extraction of ginger oleoresin with supercritical carbon dioxide, Ind. Eng. Chem. Res. 42 (5) (2003) 10571063. [24] X. Zhang, W.T. Iwaoka, A.S. Huang, S.T. Nakamoto, J.M. Wong, Gingerol decreases after processing and storage of ginger, J. Food Sci. 59 (6) (1994) 13381343. [25] J.P. Bartley, P. Foley, Supercritical uid extraction of Australian grown ginger (Zingiber Ofcinale), J. Sci. Food Agric. 66 (1994) 365371. [26] J.P. Bartley, A.L. Jacobs, Effects of drying on avour compounds in Australian-grown ginger (Zingiber Ofcinale), J. Sci. Food Agric. 80 (2000) 209215. [27] K.C. Zancan, M.O.M. Marques, A.J. Petenate, M.A.A. Meireles, Extraction of ginger (Zingiber Ofcinale Roscoe) oleoresin with CO2 and co-solvents: a study of the antioxidant action of the extracts, J. Supercrit. Fluids 24 (2002) 5776.

indeed a pulp or slurry is likely to result. The interfacial area for mass transfer between this fresh ginger slurry and the supercritical uid could be reduced relative to the dried ginger case.

5. Conclusion This work has shown that the use of fresh ginger for supercritical extraction provides a greater yield than the oven or air-dried feed currently employed commercially. When the moisture levels within the fresh ginger are high as is the case in new season rhizomes, the extract yield is also enhanced relative to a freeze-dried feed. However, the yield enhancement falls signicantly when the drier and more brous old harvest ginger is utilised. Mathematical modelling of the experimental data shows that these changes are principally related to changes in the effective diffusivity of the extract within the solid matrix. Further work might consider whether mass transfer rates in old season ginger could be enhanced by pre-soaking this material to increase the water load. The use of fresh ginger for supercritical extraction also leads to a product that is richer in the pungent gingerols than the oven dried feed commonly used. The literature suggests that this extract could be better avoured and more pungent, which could be of signicant marketing advantage commercially. However, there are also some manufacturing disadvantages relating to the use of a high volume and perishable feed stream that would need to be considered.

Acknowledgements Financial support for this project from the Co-operative Research Centre for Bioproducts is gratefully acknowledged. Mr. Balachandran receives an International Postgraduate Research Scholarship from the Australian government and a Melbourne Research Scholarship from the University of Melbourne and this support is further gratefully acknowledged. The infrastructure support for this project from the Particulate Fluid Processing Centre, a Special Research Centre of the Australian Research Council, is also gratefully acknowledged.

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