Journal of Food Engineering 76 (2006) 128138 www.elsevier.

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Production of rectied grape musts by ion exclusion chromatography

Vu Hong Thang
a

a,b

, Senad Novalin

a,*

Department of Food Science and Technology, University of Natural Resources and Applied Life Sciences (BOKU), Muthgasse 18, A-1190 Vienna, Austria b Department of Fermentation Technology, Hanoi University of Technology, 1 Dai Co Viet Road, Hanoi, Vietnam Received 22 December 2004; accepted 27 April 2005 Available online 6 July 2005

Abstract Traditionally, rectied grape musts were produced by passing the claried juice over a series of cation and anion exchange resins to remove salts, organic acids and nitrogenous materials. This process has lower productivity and causes environmental issues due to the requirements of regenerating ion exchangers. In order to solve these problems, a new process for the production of rectied concentrated grape musts from grape juice was proposed. Instead of using conventional rening processes, e.g., ion exchange-based method, rectied grape musts could be simply produced in a single step of ion exclusion chromatography. The eect of ow rate and loaded volume of feed on the separation performance was investigated. Grape sugars were easily separated from inorganic salts as well as organic acids. High purity (more than 99.9%) of rectied must was obtained with high recovery yield (approximately 99.0%). Organic acids were not detected in the rectied must while total content of inorganic ions was lower than 0.70 g/kg of total sugars. The amount of total polyphenolic compounds varied from 47.1 to 104.7 mg/kg total sugar (as mg of gallic acid equivalent). However, the eect of cations other than potassium on the separation performance requires a long-time investigation. 2005 Elsevier Ltd. All rights reserved.
Keywords: Grape juice; Rectied grape musts; Preparative chromatography; Ion exclusion; Desalting

1. Introduction To produce a high-quality wine, it is important to obtain a ne balance between the various chemical constituents, especially between the sugars and acid content. Adjustment of the must prior to fermentation enables the winemaker to start the fermentation with all juice components in balance. When the grape musts do not have a sucient potential alcohol content, it is necessary to increase their sugar concentration by enrichment, which plays an important role in wine production. Enrichment is an oenological practice, which is used to increase the natural alcoholic strength of a wine. Grape must is enriched by adding sugar-containing products,
Corresponding author. Tel.: +43 1 360066288; fax: +43 1 360066250. E-mail address: senad.novalin@boku.ac.at (S. Novalin). 0260-8774/$ - see front matter 2005 Elsevier Ltd. All rights reserved. doi:10.1016/j.jfoodeng.2005.04.055
*

which may be sucrose or grape musts (concentrated must or rectied concentrated must). Rectied concentrated must (RCM) is a concentrated solution of whole grape sugar, used for the natural enrichment of wines. RCM is the ideal product for this enrichment because it respects the particularities and the aromatics as well the identity of every variety of wine grapes. Containing only grape sugar, RCM does not change the characteristics of wine to which it is added. RCM can also be used to sweeten wines and for fermentation in the production of lightly bubbly and sparkling wines. It is a product of identical composition to grapes as opposed to sucrose. The latter although when added produces the same sweetening eect it remains an outside element to the composition of wine, being a derivative of the beetroot or cane. In wine contests, comparison between wines where RCM was used towards the others upgraded by sucrose, typically favours

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the former. Moreover, the use of RCM is permitted to all wine-makers within EU countries while the users of it enjoy the communitarian subsidies in accordance with regulations pertaining to the upgrading process. Enrichment using sucrose is a technique applied traditionally in a large number of wine-growing regions in the centre and north of the EU countries. In these countries, the governments have to pay considerable amount to support the wine makers who use RCM or rectied musts for enrichment (Commission Regulation (EEC) No. 1059/83 and (EC) No. 625/2003). However, in the south countries, like Italy, the use of sucrose in wine enrichment is forbidden (Commission Decision 89/228/EEC). It was agreed to uphold the ght against the use of sucrose, which should be replaced by enrichment with rectied concentrated must. However, due to the higher cost than sucrose, this idea is still under discussion. It is believed that if the price of RCM is reduced, the use of sucrose for enrichment could be prohibited. On the other hand, wine-growing countries in the EU have often an over-production amount of grape each year, which is being used to produce low-quality wines and other low-price products. For more economic aspects as well as for a sustainable development, an ecient utilization of this amount has to be taken into account. The RCM is a product currently obtained from grape must subjected to rectication by passing the claried juice over a series of cation and anion exchange resins to remove salts, organic acids and nitrogenous materials. The resultant juice feed also may be passed over an adsorbent resin or carbon bed to remove colour and reduce odours (Junge, 1986). The impurities are stated by CEE regulations No. 8822/87 and No. 2253/98 (Pirrone, 2000). The obtained liquid is then concentrated in a falling lm, multi-stage plate evaporator functioning under vacuum at low boiling temperatures, to produce grape sugar syrup of approximately 6870 Bx. The product obtained is a clear, uncolored or yellowish liquid, containing glucose and fructose in the same proportion as in the original must, and traces of other constituents (Pompei, 1982). With this process, the big disadvantage is that the ion exchange resins require regeneration and therefore a lot of reagents will be released. Besides ion exchangers, electrodialysis was also tested for removal of ionic compounds (Bidan, 1994; Escudier et al., 1985). Chromatographic separation is a widely used unit operation in sugar or sugar derivative production. The rst industrial batch systems were built in 1960s and 1970s. In the mid 1980s, the continuous simulated moving bed (SMB) process was applied in separation of molasses. The sequential SMB process, which can simultaneously recover multiple product fractions, was introduced in the late 1980s (Paananen & Kuisma, 2000). In sugar and other industries, ion exclusion chromatography is a proven economical alternative to the use of ion exchange resins for the separation of sugars from

non-sugars. The sugar compounds or other compounds to be puried, usually non-ionized, are adsorbed on the surface of the resins and recovered by elution. The resin is chosen to have the same charge as the impurities that are therefore not retained. Several of the many advantages of chromatography can be mentioned such as reduction of the euent volume and of the pollution load, avoidance of chemical regenerants or a signicant decrease of energy consumption. However, unlike molasses or sugar syrups, the pH of grape juice is quite acidic (pH = 3.54.0), therefore the performance separation is still not clear yet. In this work, production of rectied grape must using ion exclusion chromatography was preliminarily investigated. Some important operating parameters such as ow rate of eluent and loaded volume of feed were discussed.

2. Materials and methods 2.1. Grape must White grape was pressed and the pressed juice then was microltered by 0.2 lm polypropylene membrane (MicrodynNadir, Germany). The concentrated juice was obtained by evaporating the microltered juice in vacuum condition (60 C, 80150 mmHg). All samples were stored in a frozen room at 30 C for further use. The compositions of grape juices are shown in Table 1. 2.2. Adsorption isotherms The adsorption isotherm was determined by using a 1:10 (w:v) ratio of resin and starting solution (Tong
Table 1 Composition of grape must and concentrated grape must Parameter Bx Conductivity, mS/cm pH Glucose, g/l Fructose, g/l Total polyphenolic compounds, mg/l (as gallic acid equivalent) Tartrate, g/l Malate, g/l Citrate, g/l K+, g/l Na+, g/l NH , g/l 4 Ca2+, g/l 2+ Mg , g/l SO2 , g/l 4 PO3 , g/l 4 Grape must 15.0 3.15 4.00 75.20 76.30 223.10 Concentrated grape must 37.0 7.50 3.90 201.78 203.99 479.20

2.10 2.50 0.20 2.60 0.03 0.07 0.09 0.05 1.23 0.65

5.64 6.47 0.50 7.32 0.07 0.15 0.01 0.01 3.38 1.69

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et al., 2004). Aqueous solutions containing glucose and fructose at dierent concentration were used as starting solutions. Finally, all vials containing the resin and starting solutions were placed for 24 h in a water bath at 60 C. Sugar concentrations in the liquid phase at equilibrium condition were measured by HPLC and then, the amount of adsorbed sugars per gram of wet resin was calculated using the following equation: C C 0 C E V W 1

where C* is the amount of sugar adsorbed by the resin (g/g dry resin); C0 is initial concentration of glucose/ fructose in starting solution (g/l); CE is glucose/fructose concentration at equilibrium (g/l); V is initial volume of starting solution (L); and W is weight of resin (g). 2.3. Preparative chromatography

backwashed with reverse osmosis water to remove ne particles. The air trapped in the resin was removed and the plunger was lowered to the top of the resin bed. This step was followed by downwardly washing with water until the pH at the outlet of column was similar to that of water. After the preparation of column, chromatographic separation was carried out. The feed solution was introduced into the column via a sample loop or a peristaltic pump. A constant temperature circulator was used to maintain the temperature (at 60 C) of water in the jacket. Reverse osmosis water was used as eluent, which was downwardly pumped. Fractions were automatically collected by an automatic sample collector and analyzed later. Conductivity and pH of euent were monitored online. The separation process was controlled by a computer with software PrepCon. 2.4. Determination of dead volume

A pilot scale preparative chromatography unit made by Gold Fish GmbH, Austria (Fig. 1) was used. The jacketed 100 2.5 cm (inner diameter) column from Kronlab (Sinsheim, Germany) equipped with adjustable plungers was charged with the cation exchange resin Finex CS16 GC in potassium form (Finex Oy, Finland). The resin bed volume (BV) was 0.45 l. Resin was mixed with water treated by reverse osmosis for 24 h at room temperature, and oating resins were removed. Resins then were packed into the column. The resin bed was

The dead volume was determined by placing 10 ml of 1 g/l Blue Dextran (molecular weight of approximately 2,000,000) solution at the top of the column, allowing material to ow through the column at a xed ow rate of 2.04 cm/min. The signal was measured at the absorbance of 620 nm and the time required for passing Blue Dextran through the column was recorded. From the obtained data, the dead volume was calculated to be 180 ml (0.4BV).

Fig. 1. Pilot-scale preparative chromatography system.

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2.5. Analyses Glucose and fructose were analyzed by HPLC (Dionex Submit, USA) equipped with a RI detector and an Aminex HPX-87C column (Bio-Rad, CA, USA). The column temperature was maintained at 85 C by using a column oven. The mobile phase was 2.5 mM CaCl2 at a ow rate of 0.7 ml/min. Organic acids were analyzed by HPLC (Dionex Submit, USA) equipped with a RI detector and an Aminex HPX-87H column (Bio-Rad, CA). The column temperature was maintained at 65 C by using a column oven. The mobile phase was 10 mM H2SO4 at a ow rate of 0.6 ml/min. The concentration of inorganic anions was analyzed using ion chromatography (Dionex DX-120, USA) with ED40 conductivity detector, IonPac AS 14 column using NaHCO3/Na2CO3 1 mM/3.5 mM at 1.2 ml/min, 25 C as mobile phase. The concentration of inorganic cations was analyzed using ion chromatography (Dionex DX120, USA) with ED 40 conductivity detector, IonPac CS 12A column using methanesulfonic acid 20 mM at 1.2 ml/min, 25 C as mobile phase. The analysis is evaluated by the software Peaknet Control and Chromelon (Dionex, USA). Total polyphenolic compounds were measured by the FolinCiocalteu method adapted to a micro scale (Arnous, Makris, & Kefalas, 2002). In a 1.5-ml Eppendorf tube, 0.79 ml distilled water, 0.01 ml sample appropriately diluted, and 0.05 ml Folin-Ciocalteu reagent were added and vortexed. After exactly 1 min, 0.15 ml of sodium carbonate (20%) was added, and the mixture was vortexed and allowed to stand at room temperature in obscurity, for 120 min. The absorbance was read at 750 nm, and the total polyphenolic content was calculated from a calibration curve, using gallic acid as standard (50800 mg/l). Results were expressed as mg/l gallic acid equivalents (GAE).

0.20 0.16
C*. g/g of resin

Simulated equilibrium isotherms 0.12 0.08 0.04 0.00 0 50 100 150 200 250 CE. g/L

Fig. 2. Adsorption isotherms of glucose and fructose on Finex CS16GC resin (potassium form). C*amount of sugar adsorbed by the resin; CEsugar concentration in liquid phase at equilibrium; (d) experimental data of glucose; (s) experimental data of fructose.

3. Results and discussion 3.1. Equilibrium isotherms and column separation The isotherms of glucose and fructose in a mixture of these sugars were investigated. The adsorption isotherms are shown in Fig. 2. It can be seen that this is not a linear isotherm. It should be noted that the frequently made assumption of linear and uncoupled isotherms of glucose and fructose on cation exchange resin might not valid at high concentration and it aects considerably the predicted steady-state prole within a process chromatography system. This in turn leads to operational parameters that, if based on the model, result in less than optimum separation. Saska, Clarke, Wu, and Iqbai (1991) reported that the isotherms of

glucose and fructose in a mixture of these two sugars were not linear under conditions of high concentrations and the non-linearity of the isotherms was explained by considering the limited accessibility of the internal surface of the resin for the sugar molecules. The isotherms show that there was almost no separation between glucose and fructose. It can qualitatively be seen in Fig. 2 that there is no signicant dierence between the ratio of partition coecient of glucose and fructose. Contrarily, in the case of resin in calcium form the ratio of partition coecient of glucose and fructose was very high (Saska et al., 1991). Classically, separation of sugars is done with resins in the calcium form. The resins in the calcium form lead to a fructosecalcium complex while glucose does not form complex with the resin, thus the separation can easily be done. In case of resins in the potassium form, the separation occurs with the principle of ion exclusion, therefore both glucose and fructose are eluted with a similar time. The column separation was tested using microltered grape must. Fig. 3 shows a good separation between sugars and non-sugars in column separation of grape sugars. Glucose was eluted slightly faster than fructose but in practice it can be considered that both sugars were eluted together. The reason is that since the cation exchange resin is in potassium form there is almost no separation between glucose and fructose. This result agreed with that obtained in the equilibrium isotherm determination. With the elution volume of 1.0 BV for sugar fraction, the separation process is quite fast. 3.2. Eect of ow rate on separation Flow rate is one of the most important parameters in process chromatography. In practice, the throughput is maximized by increasing the ow rate to their limits. The eects of dierent ow rates on the separation

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25 Non-sugars

V.H. Thang, S. Novalin / Journal of Food Engineering 76 (2006) 128138

2

Non-sugar concentration, g/l

20 15 10 5 0 0 0.3 0.6 0.9 1.2 1.5 Bed volume Glucose Malate Phosphate Fructose Citrate Sulfate

1.6 1.2 0.8 0.4 0

40 30 20 10 0 0 0.3 0.6 0.9 1.2 1.5 Bed volume

8 6 4 2 0

Tartarate Potassium

Sugars

Non-sugars

Fig. 3. Column separation of grape sugars on Finex CS16GC resin (potassium form). Conditions: temperature = 60 C, ow rate = 10 ml/min, Vfeed = 10 ml, column dimension (Dinner H) = 2.5 100 cm.

performance of sugars were investigated. In all experiments, an amount of 0.044 BV of three-time concentrated grape must was used as feed. The feed were pumped into the top of column, and then reverse osmosis water was downwardly pumped into the column as eluent. The samples were collected in 10 ml fractions for later analysis o-line. The obtained data were pre140
Sugar concentration, g/l

sented in Fig. 4, it can be seen that the volume retentions were almost not changed but the base of the peaks was broader. This will cause a decrease in the resolution between sugar and non-sugar peaks. As shown in Table 2, when the ow rate increased, the maximum concentration decreased. The maximum concentration decreased from 120.73 to 70.49 g/l when
140 15 12 9 6 3 0 0 0.3 0.6 0.9 1.2 1.5 Bed volume

12 9 6 3

Sugar concentration, g/l

120 100 80 60 40 20 0 0 0.3 0.6 0.9 1.2 1.5 Bed volume

140

120 100 80 60 40 20 0

15

140
Non-sugar concentraion, g/l

15 12 9 6 3 0 1.5

c
S ugar c onc entration, g/l
120 100 80 60 40 20 0 0 0.3 0.6 0.9 1.2 1.5 Bed volume Sugars Non-sugars 3 0 9 6 12

Sugar concentration, g/l

120 100 80 60 40 20 0 0 0.3 0.6 0.9 1.2 Bed volume Sugars

Non-sugars

Fig. 4. Separation of sugars from non-sugars at dierent ow rates of elution. Conditions: temperature = 60 C, Vfeed = 20 ml, column dimension = 2.5 100 cm (Dinner H), ow rate = 1.02 cm/min (a); 2.04 cm/min (b); 4.08 cm/cm (c) and 6.12 cm/min (d).

Non-sugar concentration, g/l

Non-sugar concentration, g/l

Non-sugar concentration, g/l

15

Non-sugar concentration, g/l

Sugars

50

10

b
Sugar concentration, g/l

Sugar concentration, g/l

V.H. Thang, S. Novalin / Journal of Food Engineering 76 (2006) 128138 Table 2 Eect of ow rate on resolution and productivity Flow rate, cm/min 1.02 2.04 4.08 6.12
a

133

Resolution 1.34 1.12 1.04 0.87

Maximum concentration peak, g/l 120.73 94.53 79.11 70.49

Productivitya, kg/m3h 12.85 25.17 49.11 71.77

Productivity was considered as the feed volume treated per unit volume of resin per unit time.

ow rate increased from 1.02 to 6.12 cm/min, respectively. Contrarily, the productivity increased signicantly with the increase of ow rate. The calculated resolutions decreased remarkably from 1.34 to 0.87 when the ow rate increased from 1.02 to 6.12 cm/ min, respectively. However, the productivity increased signicantly as long as the increase of ow rate. It should be noted that in large-scale chromatography, the productivity is usually more important than the resolution. Table 3 shows more in detail the eect of ow rate on the separation between sugars and non-sugars. It is clear that the productivity increased proportionally with ow rate. In all runs, the ratios of glucose and fructose were close to that of original grape must (glucose/fructose = 0.99). This proved that there was no separation between grape sugars and this agreed with the results obtained in isotherm experiments. However, at the same level of recovery yield, the concentration of sugar fraction decreased with the increase of ow rate. With the yield more than 99%, this concentration decreased from 49.06 to 33.63 g/l when the ow rate increased from 1.02 to 6.12 cm/min. It should be noted that after rectifying, the demineralised grape juice will be evaporated, therefore the lower concentration of sugar fraction, the higher energy demand for concentration step would be required.
Table 3 Eect of ow rate on the separation of sugars from non-sugars Flow rate, cm/min 1.02 Yield, % 99.16 98.90 96.56 99.43 98.57 96.57 99.38 98.34 96.57 99.13 98.05 96.09 Puritya, % 99.93 99.94 99.95 99.94 99.96 99.97 99.94 99.95 99.96 99.87 99.95 99.95 Sugar concentration, g/l 49.06 52.42 59.71 44.42 49.91 56.42 36.65 40.09 44.00 33.63 34.77 39.45

Regarding the Italian standard for rectied concentrated musts, the total inorganic ions in the product must not be higher than 1.2 g/kg total sugar (Bidan, 1994; Weger, 1985). Similarly, in Austria, in order to receive the nancial aid for enrichment of wines using RCM, the total cation amount in RCM must be lower than 8 meq/kg total sugar (MerkblattBMLFUW, Austria, 2003). The results in Table 3 show that in all runs the ow rate has no signicant inuence on the exclusion of inorganic ions. In all cases, the residue of inorganic ions in sugar fraction can satisfy the above criterion. At ow rates up to 6.12 cm/min the amount of total inorganic ions was lower than 0.70 g/kg total sugar, much lower than the EU standard. The above regulations also regulated that the total organic acid content (as tartaric acid) must be lower than 1.0 g/kg total sugar (Bidan, 1994; Weger, 1985). However, in all runs, organic acids (tartaric, malic, citric acids) were not detected in sugar fractions. Another important parameter is the total polyphenolic compounds, which should be in the range of 40100 mg/kg total sugar. It can be seen that in all runs, the total polyphenolic content varied from 49.1 to 94.1 mg/kg total sugar. For production scale chromatography, the amount of interested compound that can be separated per unit time is an essential criterion. Throughput, the weight of substance isolated in a given period of time, depends on parameter such as column dimensions and eluent ow rate. In other words, resolution is not always the primordial factor in preparative chromatography (Stead, 1998). 3.3. Eect of feed volume on separation In process chromatography, the product of interest should be processed as quickly and cheaply as possible. In order to increase the productivity of process

Glucose: fructose 0.99 0.99 1.00 0.99 0.99 0.99 0.99 0.99 0.99 0.99 0.99 0.99

Dilution factor 8.3 7.7 6.8 9.1 8.1 7.2 11.1 10.1 9.2 12.1 11.7 10.3

Inorganic ions, g/kg total sugars 0.68 0.54 0.53 0.61 0.44 0.35 0.58 0.46 0.40 0.65 0.49 0.49

Total polyphenols, mg/kg total sugar 79.4 62.4 47.1 83.5 60.5 49.1 83.8 65.9 58.2 94.1 81.3 71.2

2.04

4.08

6.12

Purity was dened as total sugars (glucose and fructose) divided by the sum of sugars, inorganic and organic ions.

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chromatography, the operation of process in overloaded mode is often used. Mass overload refers to overload of a column due to total mass of sample injected and can be used to increase the throughput of a preparative chromatography separation as long as the degree of overloading is controlled. One important factor, which limits the increase of concentration, is solubility. Mass overloading can be done by overloading feed volume or increasing the feed concentration. It is well known that the usage of large concentrations is usually advantageous. Thus, in general the maximum possible concentrations are most attractive. However, there is a constraint, which is solubility. In process chromatography, the concentration of feed was increased as high as possible. In practice, the feed concentration is brought up to the level, which is about 10% less than the solubility range because a chromatographic enrichment occurs during the separation process (Strube, Haumreisser, Schmidt-Traub, Schulte, & Ditz, 1998). In the sugar industry, the concentration of feed used for process chromatography is around 60% (w/w). In case of grape juice, such high concentration cannot be used due to the low solubility of tartrate. Since the grape juice contains a relatively high amount of potassium hydrogen tartrate, which has low solubility (6 g/l), the

grape musts could be concentrated about three-time. In this work, the sugar concentration used for feed was about 410 g/l. Because the maximal concentration of feed reached a limit, overloading volume is expected to improve the productivity. It was reported that overloading volume results when the volume of sample injected exceeds 1020% of the column dead column (Stead, 1998). In this work, the eect of feed volume on the separation performance was investigated with the same ow rate of eluent (2.04 cm/min) at dierent level 11.11%, 16.67%, 22.22% and 50.00% of the column dead volume. The results are shown in Fig. 5. It is known that the purity will be decreased as the column capacity is exceeded and solute peak shape is lost. The choice of sample loading for preparative chromatography will depend on many factors; for example, the nature of the components that are present in the sample and their chromatographic proximity to the component of interest, the purity of product required, the column capacity, and the total amount of sample to be processed. It can be found that loaded volume has a similar eect on separation like ow rate. In Fig. 5, it is obvious that the volume retentions were very slightly dierent but the base of the peaks was broader. Consequently, the separation eciency decreased due to the increasing overlapped area.

180

30

180

30

Sugar concentration, g/l

Sugar concentration, g/l

120 90 60 30 0 0 0.3 0.6 0.9 1.2 1.5 Bed volume

180

20 15 10 5 0

120 90 60 30 0 0 0.3 0.6 0.9 1.2 1.5 Bed volume

20 15 10 5 0

30

350

30

c
Non-sugar concentration, g/l S ugar c onc entration, g/l
150 120 90 60 30 0 0 0.3 0.6 0.9 1.2 1.5 Bed volume
Sugars

d
300 250 200 15 150 100 50 0 0 0.3 0.6 0.9 1.2 1.5 Bed volume 10 5 0

20 15 10 5 0

20

Non-sugars

Sugars

Non-sugars

Fig. 5. Eect of feed volume on the separation of sugars from non-sugars. Conditions: temperature = 60 C, ow rate = 2.04 cm/min, Vfeed = 20 ml (a); 30 ml (b); 40 ml (c) and 90 ml (d).

Non-sugar concentration, g/l

25

25

Sugar concentration, g/l

Non-sugar concentration, g/l

150

25

Non-sugar concentration, g/l

b
150 25

V.H. Thang, S. Novalin / Journal of Food Engineering 76 (2006) 128138 Table 4 Eect of loaded volume on resolution and productivity Loaded volume, % of column dead volume 11.11 16.67 22.22 50.00 Resolution Maximum concentration peak, g/l 94.53 122.12 153.46 289.34 Productivitya, kg/m3h 25.61 38.24 49.67 89.95

135

1.12 1.12 0.90 0.86

a Productivity was calculated as sugar amount obtained per unit volume of resin per unit time. The calculations were with the yields of around 96%.

Data in Table 4 show that when the loaded sample increased from 11.11% to 22.22%, the retention volume of sugar peak was not changed. Only when the loaded sample signicantly increased (from 22.22% to 50.00%), the retention volume slightly increased (from 0.822 to 0.889, respectively). Data in the resolution of the sugar and non-sugar peaks decreases signicantly (from 1.12 to 0.86) because increasing loading volume increases the peak width more than the retention volume. When the loaded volume was 50% of the dead volume, the sugar peak came out at a slightly higher retention volume but its width was quite a lot wider. However, it should be mentioned that in practice, overloaded volumes are often used for process chromatography. As expected, the maximum concentration as well as the productivity increased signicantly but not linearly with the increase of loaded volume. When the loaded volumes lower than 22% of the column dead volume, the productivity increased nearly linearly with the increase of loaded volume. But, at the loaded volume of 50%, this result was not observed. Data in Table 5 show in detail the separation performance. Obviously, when the loaded volume of feed was approximately lower than 22% of column dead volume,

there was no signicant dierence in the correlation between the recovery yield and the purity: the same purity levels required a similar recovery yield. However, if the loaded volume of feed increased up to 50% of column dead volume, this correlation changed signicantly. In order to have similar purities like the cases of loaded volume of feeds smaller than 22% of column dead volume, the recovery yield will be reduced from 2.5 to 8.0%. As mentioned above, the total inorganic ions in rectied musts have to be lower than the value of 1.2 g/kg total sugar (Bidan, 1994; Weger, 1985). The results in Table 5 show that, when the loaded volume increased to 22% of the column dead volume, the inorganic residues still satised the above standard. Even when the loaded volume reached 50% of the column dead volume, the inorganic ion content still satised if the recovery yield not exceeds 92%. Another important parameter is the total polyphenolic compounds, which should be in the range of 40100 mg/kg of total sugars. It can be seen from Table 5 that nearly all grape musts have the total polyphenolic compounds in above range. It is well known that, dilution is one of some disadvantages of preparative chromatography process. In order to reduce the dilution factor, either increasing the feed concentration or feed volume is often used. In the case of grape must, since the maximum concentration of feed was limited, overloading feed becomes an eective way to improve the dilution factor. It is obvious that when the loaded volume increased the concentration of sugar fraction increased remarkably, which lead to the decrease of dilution factor. The dilution factor can be lowered as much as 2.0, however the lower dilution factor is the lower ratio of glucose and fructose. At the same degree of recovery yield, the average concentration of sugar fraction increased with increasing the loaded volume. However, this increase was not proportional to increased volume.

Table 5 Eect of feed volume on the separation of sugars from non-sugars Feed volume, % of the column dead volume 11.11 Yield, % 99.43 98.57 96.57 98.84 98.20 95.84 98.96 98.12 96.17 96.92 92.07 87.94 Puritya, % 99.94 99.96 99.97 99.92 99.92 99.94 99.80 99.91 99.95 99.52 99.77 99.84 Sugar concentration, g/l 44.42 49.91 56.42 67.22 71.24 80.22 78.77 87.29 96.97 156.11 181.26 194.77 Glucose:fructose 0.99 0.99 0.99 0.99 1.00 1.00 0.98 0.98 0.96 0.95 0.91 0.88 Dilution factor 9.1 8.1 7.2 6.0 5.7 5.1 5.2 4.6 4.2 2.5 2.2 2.0 Inorganic ions, g/kg total sugar1 0.61 0.44 0.35 0.78 0.78 0.56 1.16 0.76 0.54 2.08 1.19 0.95 Total polyphenols, mg/kg total sugar 83.5 60.5 49.1 69.4 63.3 50.6 81.2 69.4 57.4 104.7 81.7 72.8

16.67

22.22

50.00

Purity was dened as total sugars (glucose and fructose) divided by the sum of sugars, inorganic and organic ions.

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3.4. Proposed process for the production of rectied concentrated must (RCM) Based on data mentioned above, a schematically proposed process like Fig. 6 can be given for the production of rectied concentrated must. Comparing with the conventional process, it can be seen that in any case, the evaporation step after rectifying the juice is inevitable. Therefore, the step for removing ions is key technology of the conventional process. In chromatography-based process (proposed process) only one step required for removing inorganic ions while in conventional process, at least cation and anion exchangers are needed. It should be noted that the more separation steps used the lower total recovery yield as well as lower productivity is. In addition, a big disadvantage of the ion exchange-based process is that after a certain time of

operation, the ion exchange resins will be exhausted. Consequently, the regeneration step is often required but much more inorganic salts will be produced as wastes. With this proposed process, the investment for equipment can be minimized because not only the ion exchange columns can be used for chromatography columns, but also other equipments, which are being used for treating juice, can be used for the new process. Only one more concentration step (evaporation 1) is required. In this step, the juice is concentrated to concentration of about 40 Bx, depending mainly on the tartrate content. After the chromatographic separation step, the sugar content of the sugar fraction could be somewhat similar to that of raw grape juices. In conventional process, it can be said that the sugar content after ion exchange steps is quite similar to that of raw juices

Fig. 6. Possibly schematic process for the production of RCM based on ion exclusion chromatography and evaporation.

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(only slightly diluted). Therefore, the total energy consumption for evaporation was estimated to be 1.51.8 fold higher than that in conventional methods (ion exchange-based process). However, this water can be used as eluent for the chromatographic separation step. The dilution problem in chromatographic separation is expected to be improved with the simulated moving bed (SMB) system. It was reported that, in case of fructose/glucose separation (in large-scale), SMB process could reduce up to 2.5 times of solvent requirement and increase the product concentration up to 2.3 times (Strube et al., 1998). Alternatively, reverse osmosis (RO) showed a good potential for concentrating grape musts (Berger, 1991; Ferrarini, Versari, & Galassi, 2001; Mietton-Peuchot, MiLisic, & Noilet, 2002). Mietton-Peuchot et al. (2002) reported that grape musts could be concentrated by RO until when the potential alcohol content reached 15, which is equal to about 300 g/l of sugars. By using RO for pre-concentration of juices, the total energy consumption for concentration/evaporation steps could be much reduced.

were eluted in the salt fraction, could further be separated by similar technology. It was known that at low pH, organic acids are in neutral form, which is easily separated from charged compounds (Kulprathipanja, 1988, 1989; Kulprathipanja & Oroskar Anil, 1991). By doing that, it is possible to recover the main valuable compounds, e.g., grape sugar, tartaric and malic acids.

References
Arnous, A., Makris, D. P., & Kefalas, P. (2002). Correlation of pigment and avanol content with antioxidant properties in selected aged regional wines from Greece. Journal of Food Composition and Analysis, 15, 655665. Berger, J. L. (1991). Auto-enrichissement du mout par osmose inverse. Bulletin de lOIV 721722, 189210. Bidan, P. (1994). Techniques delaboration du mout de raisin concentre rectie (M.C.R). Revue Francaise dOenologie, 34(147), 1720. Commission Decision of 30 November 1988 on Decree-Law No 370/87 of 7 September 1987 of the Italian Government converted into Law No 460 of 4 November 1987 on production and marketing, including new standards for the production and marketing of wine sector products. Ocial Journal L 094, 07/0471989, pp. 0038 0042. Commission Regulation (EEC) No 1059/83 of 29 April 1983 on storage contracts for table wine, grape must, concentrated grape must and rectied concentrated grape must. Ocial Journal L 116, 30/04/1983, pp. 00770083. Commission Regulation (EC) No 625/2003 of 2 April 2003 amending Regulation (EC) No 1623/2000 laying down detailed rules for implementing Council Regulation (EC) No 1493/1999 on the common organisation of the market in wine with regard to market mechanisms. Ocial Journal L 090, 08/04/2003, pp. 0004 0031. Escudier, J. L., Moutonnet, M., Barillere, J. M., Benard, P., Cottereau, P., Audinos, R., et al. (1985). Lelectrodialyse: demineralisation et desacidication des mouts de raisin. Fabrication de mout con centres recties. Revue Francaise dOenologie, 25(99), 3944. Ferrarini, R., Versari, A., & Galassi, S. (2001). A preliminary comparison between nanoltration and reverse osmosis membranes for grape juice treatment. Journal of Food Engineering, 50, 113116. Junge, von C. (1986). RTK-Herstellungsverfahren, Zusammensetzung, Kontrollmoglichkeiten. Die Weinwirtschaft/Technik, 3, 8688. Kulprathipanja, S. (1988). Separation of citric acid from fermentation broth with a neutral polymeric adsorbent. US Patent, US4720579. Kulprathipanja, S. (1989). Separation of citric acid from fermentation broth. European Patent, EP0324210. Kulprathipanja, S., Oroskar Anil, R. (1991). Separation of lactic acid from fermentation broth with an anionic polymeric absorbent. US Patent, US5068418. Merkblatt: Beihilfe fur die Anreicherung mit rektiziertem Trauben mostkonzentrat (RTK) bzw. Konzentriertem Traubenmost (konz. TM). Austrian Federal Ministry of Agriculture, Forestry, Environment and Water Management (BMLFUW). Available from: http://www.lebensministerium.at/lebensm/ (accessible in December 2004). Mietton-Peuchot, M., Milisic, V., & Noilet, P. (2002). Grape must concentration by using reverse osmosis. Comparison with chaptalization. Desalination, 148, 125129. Paananen, H., Kuisma, J. (2000). Chromatographic separation of molasse components. In Proceedings of 7th AVH association aymposium, Reims.

4. Conclusions A quite simple technique was developed for rectied grape must production. The eect of ow rate and loaded volume of feed on the separation performance was investigated. The results showed that grape sugar was easily separated from inorganic salts as well as organic acids. Some important parameters were satised with the current standards for RCM. High purity (more than 99.9%) of rectied musts was obtained with high recovery yield (approximately 99.0%). Organic acids were not detected in the rectied must while total content of inorganic ions was lower than 0.70 g/kg total sugar. The amount of total polyphenolic compounds varied from 47.1 to 104.7 mg/kg total sugar (as mg of gallic acid equivalent). Based on the obtained data, a process was proposed for RCM production. This process shows a great potential for industrial-scale production of RCM. From the viewpoint of sustainable production, this process is better than the conventional one because much lesser chemical euents are released. Regarding the present situation of RCM production, this new process is expected not to require signicant addition of investment cost because all equipments, which are being used for ion exchange-based process, can be used for chromatography-based process. However, it is suggested that the eects of cations other than potassium on the lifetime of cation exchange resins should be investigated in detail. Besides the grape sugar fraction, a non-sugar fraction, which contains mainly organic acids and inorganic salts, is also obtained. Tartaric and malic acids, which

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V.H. Thang, S. Novalin / Journal of Food Engineering 76 (2006) 128138 Strube, J., Haumreisser, S., Schmidt-Traub, H., Schulte, M., & Ditz, R. (1998). Comparison of batch elution and continuous simulated moving bed chromatography. Organic Process Research & Development, 2, 305319. Tong, W. Y., Fu, X. Y., Lee, S. M., Yu, J., Liu, J. W., Wei, D. Z., et al. (2004). Purication of L-(+)-lactic acid from fermentation broth with paper sludge as a cellulosic feedstock using weak anion exchanger Amberlite IRA-92. Biochemical Engineering Journal, 18(2), 8996. Weger, B. (1985). Analysenwerte handelsublicher rektizierter Trau benmostkonzentrate. Weinwissenschaft, 40(3), 202205.

Pirrone, L. (2000). Method of solidifying concentrated rectied must and sugar-solutions. European Patent EP 1096006 A1. Pompei, C. (1982). Grape sugar. Technological aspects (production and utilization). Bulletin de lOIV 55, 2552. Saska, M., Clarke, S. J., Wu, M. D., & Iqbai, K. (1991). Application of continuous chromatographic separation in the sugar industry. Part I: glucose/fructose equilibria on Dowex Monosphere 99 CA resin at high sugar concentrations. International Sugar Journal, 93, 223228. Stead, P. (1998). Isolation by preparative chromatography. In R. J. P. Cannell (Ed.), Natural products isolation. Totowa, NJ: Humana Press Inc.