2nd Mercosur Congress on Chemical Engineering

th
4 Mercosur Congress on Process Systems Engineering

ASSESSMENT OF ULTRAFILTRATION AND VACUUM

EVAPORATION ON WHEY PROTEIN CONCENTRATION
L. Serpa1, F.F. Rauber1, A.J. Cichoski1 and M. Di Luccio1*
1
Programa de Mestrado em Engenharia de Alimentos – Universidade Regional Integrada do Alto Uruguai e das Missões

Abstract. In Brazil, unlike the rest of the world, cheese whey is still considered a residue of low importance
on the nutritional point of view, and is often directed to animal feeding, to wastewater treatment of dairy plants or
simply discharged in the environment. In the USA, 90% of cheese production residue is used in manufacturing of
ingredients destined to human nutrition, which meant more than 500,000 tons of whey in 2001. From 1998 to
2001, Brazil imported more than 140,000 tons of dry whey, due to the lack of internal production. Low pressure
evaporation can minimize drastically operation costs of the concentration process, since cheese whey contains
more than 93% of water. Many protein fractions of milk are thermally sensitive and the increase in temperature
may also favor reactions of proteins with sugars present in the whey. Therefore, vacuum evaporation can reduce
protein losses caused by long exposure of whey to high temperatures. Ultrafiltration has also been used in many
applications in the dairy industry. Such separation process presents an additional advantage over vacuum
evaporation because it allows whey proteins to be concentrated without concentrating lactose that permeates
ultrafiltration membranes. The recovery of proteins from whey would help to reduce the organic load discharged
by dairy industry in the environment and also to obtain a high value product that can be fractioned and used as
ingredient in special food products. In this context, the present work aimed to compare the concentration and
recovery of cheese whey albumin by ultrafiltration and vacuum evaporation processes, evaluating the effect of
process operation parameters on the quality of the protein concentrate obtained.

Keywords: whey protein, vacuum evaporation, ultrafiltration.

1. Introduction

In cheese manufacturing usually a large amount of whey is produced. This byproduct contains low amount of
fat, minerals, hydrosoluble vitamins and proteins. The whey proteins include -lactoalbumin and globulins, which
show molar mass around 17,000 Daltons and 170,000 Daltons, respectively. These proteins are highly digestible
and thus adequate for human nutrition during early age and for patients with digestion disturbances (Sgarbieri,
1996).
Currently approximately 50% of cheese whey in the world is applied in production of fermented beverages,
juices, bakery, used in animal nutrition and is usually dehydrated e commercialized as energy and nutritive
supplement (Almeida et al, 2001).
Unlike the rest of the world, cheese whey still is considered as waste in Brazil. In USA a large amount of
dairy industry wastewaters are recovered and 90% cheese production residues are used in manufacturing of
products for human nutrition, reaching more than 500,000 tons in 2001 (USDEC, 2004). From 1998 to 2001,
Brazil imported more than 140,000 tons of dry cheese whey, due to the lack of internal production (Almeida et
al., 2001; Pinheiro et al., 1993). Evaporation at low pressure can minimize operation costs, since cheese whey
contains more than 93% of water (Linden, 1996). Besides, some proteins fractions may be damaged by exposure
to high temperatures and can also react with the sugars present in the whey. In the case of cheese whey, protein

*
To whom all correspondence should be addressed.
Address: Av. Sete de Setembro 1621, Erechim, RS – Brazil

1
 2nd Mercosur Congress on Chemical Engineering
th
4 Mercosur Congress on Process Systems Engineering

fractions may present different stabilities to temperature. The most thermally resistant is -lactoglobulin,
followed by -lactoglobulin and immunoglobulins.
Ultrafiltration has also been used in many applications in the dairy industry (Bassetti, 2003). Like other
membrane separation processes it offers many advantages over conventional technologies. Its application in food
industry is particularly interesting since membrane processes can operate at mild temperatures, avoiding damages
caused by thermal processes, thus, maintaining the original characteristics of the processed products (Bronstein,
1998). Ultrafiltration presents an additional advantage over vacuum evaporation because it allows whey proteins
to be concentrated without concentrating lactose that permeates ultrafiltration membranes.
In this context, the objective of this work was to evaluate the concentration of protein of mozzarella cheese
whey by vacuum evaporation and ultrafiltration, aiming to recover high value products from this dairy industry
residue and reduce its discharge in receiving bodies.

2. Materials and Methods

2.1. Cheese whey

Mozzarella cheese whey was obtained from a local dairy industry. The whey was collected diary and kept at
4°C until use. Each sample was analyzed for pH, titrable acidity (°Dornic), total and soluble (°Brix) solids,
lactose, protein and color.

2.2. Vacuum evaporation

A vacuum evaporator (“Stephan Geiger” 3678, model UMMSK-12), consisting in a 8 Liter chamber with
temperature control through a partial vapor jacket with manual valve for controlling temperature and pressure.
The stirring system was equipped with two stirrer blades at 2 and 4 cm from the bottom of the chamber and a
digital control.
For evaluation of the effect of temperature, pressure and agitation a complete 23 factorial design was
performed. The range of the investigated variables was determined after some preliminary experiments and is
presented in Table 1. Each evaporation experiment was carried out for 30 minutes. The responses were analyzed
the concentration factor, protein, lactose, total solids in function of the factors in Table 1. Central points were
performed in triplicate to evaluate non-linearity and experimental error (Chang et al., 2002). The data were
treated with the aid of Statistica 5.0 (Statsoft Inc., Tulsa, OK, USA). All analyses were performed considering a
level of 95% of confidence (p<0.05).
After the experiment a sample of 100 mL of the concentrate was taken and used in physico-chemical
determinations. Of each sample were analyzed: pH, acidity (Tronco, 1997), density (Behmer, 1996), °Brix, total
solids (Tronco, 1997), lactose (Silva, 1997), protein by Kjedahl method (AOAC, 1975) and colorimetric
(Bradford, 1976) and color using a Minolta CR 400 colorimeter.

E-mail: diluccio@uricer.edu.br
2
 2nd Mercosur Congress on Chemical Engineering
th
4 Mercosur Congress on Process Systems Engineering

Table 1. Values of coded levels and real values used in the complete factorial design (23) used to evaluate cheese whey
concentration by vacuum evaporation

Independent Coded Levels

Variables -1 +1 0*
Temperature (°C) 40 50 45
Manometric Pressure
-0.4 -0.8 -0.6
(kgf.cm-2)
Stirring rate (rpm) 170 230 200
* Central Point

2.3. Ultrafiltration

Ultrafiltration experiments were carried out in a 100 L pilot plant at Senai-Chapecó (SC, Brazil). The plant
consists in a tangential hollow fiber ultrafiltration module with area 4.4 m2 and molecular weight cut-off
(MWCO) of 10,000 Daltons. The UF unit is equipped with temperature, pressure and flow rate controllers.
The effect of temperature, pressure and flow rate on permeate flux and physico-chemical characteristics of
permeate were evaluated using a 23 complete factorial experimental design. Central points were here also
performed in triplicate to evaluate non-linearity and experimental error (Chang et al., 2002). The range of the
factors investigated in this experimental design was chosen based on limitations of the equipment and are
presented in Table 2.
Each experimental run began and ended with a 60 min. clean water permeation with total recycle of permeate
stream. The flux was followed during regular time intervals. After clean water permeation the system was purged
and then fed with cheese whey. 60 min permeation with total recycle was carried out, followed by a 6-fold
concentration. Flux was also followed in whey permeation and samples were collected for physico-chemical
analyses.
Module cleaning was carried out right after each experiment using a CIP system. The CIP consisted in a triple
flush with clean water for 10 min each, followed by recirculation of a 6 wt% aqueous NaOH solution during 60
min. A new triple water rinsing was carried out, followed by recirculation with a 0.01 wt% aqueous solution of
H3PO4.

Table 2. Values of coded levels and real values used in the complete factorial design (23) for evaluation of UF
concentration of cheese whey

Independent Coded Levels

Variables -1 +1 0*
Temperature (°C) 40 50 45
Manometric Pressure
-0.4 -0.8 -0.6
(kgf.cm-2)
Stirring rate (rpm) 170 230 200
* Central Point

3
 2nd Mercosur Congress on Chemical Engineering
th
4 Mercosur Congress on Process Systems Engineering

3. Results and Discussion

3.1. Concentration by vacuum evaporation

Table 3 presents the matrix of the experimental design with the real and coded variables and the respective
responses. The responses that were analyzed statistically were the concentration factor and protein content. Table
4 shows the results of lactose and color of each experimental run. In concentration by vacuum evaporation both
protein and lactose are concentrated and little change could be observed in the ratio protein/lactose.
The concentrated samples also showed color variation in relation to the raw whey. It was not possible to
correlate the color variation (∆E) with lactose or protein content. However, when the color parameter is
separately compared with protein content in the concentrates, a correlation of color with protein could be
observed.

Table 3. Matrix of the complete 23 factorial design (real and coded variables)

Ratio
Run T(a) (°C) P(b) (kgf/cm2) SR(c) (rpm) CF(d) Proteína(e) (%)
Ptn/Lactose
01 40 (-1) -0.4 (-1) 170 (-1) 1.0059 12.30 0.15
02 40 (-1) -0.4 (-1) 230 (+1) 1.1877 11.83 0.16
03 40 (-1) -0.8 (+1) 170 (-1) 1.0018 12.76 0.15
04 40 (-1) -0.8 (+1) 230 (+1) 1.2121 12.21 0.15
05 50 (+1) -0.4 (-1) 170 (-1) 1.0032 12.58 0.15
06 50 (+1) -0.4 (-1) 230 (+1) 1.0874 12.71 0.14
07 50 (+1) -0.8 (+1) 170 (-1) 1.0482 7.52 0.14
08 50 (+1) -0.8 (+1) 230 (+1) 1.2240 12.26 0.15
09(a) 45 (0) -0.6 (0) 200 (0) 1.0125 11.90 0.16
09(b) 45 (0) -0.6 (0) 200 (0) 1.0813 11.86 0.14
09(c) 45 (0) -0.6 (0) 200 (0) 1.0668 12.62 0.14
Note: atemperature, bpressure, cstirring rate, dconcentration factor, eKjedahl

Table 4. Lactose and color results of each concentrate obtained in experimental design.

Lactose
Run ∆L*(a) ∆a*(b) ∆b*(c) ∆E(d)
(%)
01 5.34 -4.81 0.43 -1.73 5.13
02 5.08 -2.71 0.59 -0.31 2.79
03 5.62 -1.19 -0.08 -0.59 1.33
04 5.79 -4.36 0.38 -0.65 4.42
05 5.68 -1.06 -0.28 -0.22 1.12
06 5.75 -0.16 -1.29 -1.09 1.69
07 6.55 -0.68 -0.36 -0.17 0.79
08 6.06 -1.02 -1.68 -1.03 2.22
09(a) 5.24 -0.54 -0.35 0.15 0.66
09(b) 5.52 0.98 -0.46 -0.30 1.12
09(c) 5.72 0.90 -0.67 -0.17 1.13
Nota: (a) lightness difference; (b)red-green difference, (c) yellow-
blue difference, (d) total color variation ∆E = ∆L + ∆a + ∆b
2 2 2

4
 2nd Mercosur Congress on Chemical Engineering
th
4 Mercosur Congress on Process Systems Engineering

Figure 1 shows the Pareto chart of effects of the factor on concentration factor. It may be observed that stirring
rate presented significant (p<0.05) positive effect. Operating pressure and temperature did not show significant
effects on concentration factor, possibly due to high experimental error observed (Table 3).
p=,05

(3)Stirring ratl
22,36251

(2)Pressure
6,922293

(1)TEMP
-1,53267

-5 0 5 10 15 20 25

Effect (absolute value)

Figure 1. Pareto chart of effects of manipulated variables on concentration factor.

Variability of the raw whey

Each experimental run used a different batch of whey collected at the local dairy industry. This way, it was
necessary the monitoring of physico-chemical characteristics of each whey sample. Table 5 presents the results
obtained in this characterization, showing also the means and standard deviations of the raw material used in this
step of the study.
In the present study the water content varied from 93.1 to 94.3%, total solids between 6.0 to 6.1%, lactose
between 4.82 and 5.43% and protein between 0.77 to 0.91 (Table 6). This values are in same the range of the
characterizations performed by Veisseyre (1988), Linden (1996), Tronco (1997) and Farro (2003). It is worth to
note that the variations between the different samples were mostly below 10%.
Table 5. Physico-chemical evaluation of raw whey samples used in each evaporation experiment.
Ratio
Water TS Lactose º
Run pH ºD Density Protein1 Protein2 Ptn(NKT)
(%) (%) (%) Brix
/Lactose
1 93.5 6.5 6.58 12.0 4.8 8.90 1.02 0.81 0.21 0.17
2 94.3 5.7 6.87 11.0 5.4 9.00 1.02 0.80 0.21 0.15
3 93.5 6.5 6.58 12.0 4.8 8.90 1.02 0.83 0.25 0.17
4 94.3 5.7 6.87 11.0 5.4 9.00 1.03 0.84 0.25 0.16
5 93.1 6.9 6.85 14.0 4.9 9.40 1.03 0.88 0.22 0.18
6 94.0 6.0 6.92 12.0 4.9 9.20 1.03 0.83 0.22 0.17
7 93.1 6.9 6.71 12.5 4.8 9.50 1.03 0.91 0.34 0.19
8 94.0 6.0 6.92 12.0 4.9 9.20 1.03 0.90 0.31 0.18
9(a) 93.1 6.9 6.85 14.0 4.9 9.40 1.03 0.84 0.16 0.17
9(b) 94.0 6.0 6.92 12.0 4.9 9.20 1.03 0.77 0.21 0.16
9(c) 94.0 6.0 6.92 12.0 4.9 9.20 1.03 0.81 0.20 0.16
Average 93.7 6.3 6.82 12.2 5.0 9.17 1027 0.84 0.24 0.17
Std. Dev. 0.5 0.5 0.13 1 0.2 0.21 1.21 0.04 0.05 0.01
% Std. Dev. 0.5 7.8 1.9 8.0 4.4 2.29 0.12 4.76 20.83 5.88
Note: 1NKT (%), 2Bradford (%)

3.2. Concentration by ultrafiltration

5
 2nd Mercosur Congress on Chemical Engineering
th
4 Mercosur Congress on Process Systems Engineering

Table 6 presents the results of the factorial experimental design carried out to evaluate the effects of process
variables on permeate flux with and without recycle and the time required to concentrate the feed 6-fold. The
experimental condition that presented the higher flux with recycle was the one operated at higher pressure and
higher flow rate, while for concentration the best condition was run 4, carried out at lower temperature. A
statistical analysis of the results of the experimental design will be presented further in this paper.

Table 6. Matrix of the complete 23 factorial design (real and coded variables) and the response in term of stabilized
permeate flux
Flow Rate Flux with Concentration Flux w/o
Run T (°C) P (kgf/cm2) time (min)
(L/min) recycle** recycle**
01 25 (-1) 0.5 (-1) 45 (-1) 9.95 93.0 6.14
02 25 (-1) 0.5 (-1) 65 (+1) 6.65 90.0 4.77
03 25 (-1) 1.5 (+1) 45 (-1) 14.45 43.2 11.86
04 25 (-1) 1.5 (+1) 65 (+1) 18.08 31.5 15.59
05 45 (+1) 0.5 (-1) 45 (-1) 9.52 68.1 8.59
06 45 (+1) 0.5 (-1) 65 (+1) 9.95 66.1 6.14
07 45 (+1) 1.5 (+1) 45 (-1) 13.25 46.1 12.55
08 45 (+1) 1.5 (+1) 65 (+1) 19.80 38.5 11.73
09* 35 (0) 1.0 (0) 55 (0) 13.55 45.3 11.36
*Central point performed in triplicate.
**Stabilized flux. L/h/m2.

Figure 2 presents the evolution of permeate flux with time of permeation for the 11 runs carried out with total
recycle of permeate stream. It could be noted the reduction of permeate flux, possibly due to membrane fouling.
It is worth to note that in such cases the feed concentration remained constant and the effect of this factor does
not influence the results. In most of the runs flux is rather stable after 20 minutes of permeation. The most critical
conditions for flux decrease were the ones operated at higher pressures, when the effect concentration
polarization (accumulation of solids at membrane surface) is stronger.
The results of statistical analysis of Table 6 can be found in Figure 3. The Pareto chart shows that pressure,
flow rate and their interaction presented a significant (p<0.05) positive effect on permeate flux, while
temperature did not significantly influenced the response. This result confirm the expected since flux is directly
proportional to pressure, which is the driving force to the process, and a high flow rate can decrease the
polarization layer, decreasing mass transfer resistance.
Figure 4 shows the permeate flux during concentration runs at the central point. A further decrease in flux
could be observed when compared to the experiment runs carried out with total recycle, due to the increase in
feed concentration that causes higher polarization concentration effect.

6
 2nd Mercosur Congress on Chemical Engineering
th
4 Mercosur Congress on Process Systems Engineering

30

25 30
Permeate flux (L/h/m 2)

25

Permeate flux (L/h/m2)

20 Run 1
Run 2 20 Run 5
15 Run 3 Run 6
Run 4 15 Run 7
10
Run 8
10
5
5

0 0
0 10 20 30 40 50 60 70 0 10 20 30 40 50 60 70
Time (min) Time (min)

(a) (b)

25
Permeate flux (L/h/m2)

20

15 Run 9
Run 10
10 Run 11

0
-10 10 30 50 70
Time (min)

Figure 2. Permeate flux of cheese whey with complete recycle of permeate: (a) 25°C, (b) 45°C and (c) 35°C.

p=,05

(2)PRES 12,25361

2by3 5,41883

(3)VAZÃO_AL 3,035375

1by3 2,761319

(1)TEMP 1,40765

1by2 -,975805

-2 0 2 4 6 8 10 12 14
Efeitos absolutos

Figure 3. Pareto chart of effects of factors on cheese whey permeate flux.

16
14
Permeate flux (L/h/m2)

12
Run 9
10 Run 10
8 Run 11
6 Run 12
Run 13
4
2
0
0 10 20 30 40 50 60
Time (min)

7
 2nd Mercosur Congress on Chemical Engineering
th
4 Mercosur Congress on Process Systems Engineering

Figure 4. Permeate flux of cheese whey during concentration at 35°C.

Table 7 presents the ratio protein/lactose in the raw whey and UF-concentrated whey (mean value of all
experiments). While in vacuum concentration this ratio became almost constant after concentration, here it
increases more than 3 times. This results is in accordance to Farro (2003) that also describes na increase in this
ratio in membrane concentration using UF.

Table 7. Ratio protein/lactose for UF concentration of whey

Raw whey UF Concentrated whey Permeate
Ratio Pt/Lact 0.14 0.63 0.04
% Variation 341

4. Conclusions
Cheese whey concentration was evaluated using two different processes. In vacuum evaporation the condition
that achieved higher concentrate factor was at 50°C, -0.8 kgf/cm2 and 230 rpm, i.e., upper level of temperature,
stirring rate and vacuum.
In ultrafiltration of whey the condition that yielded higher concentration in a short time and highest permeate
flux was at room temperature, 65 L/min and 1.5 kgf/cm2.
The results show that vacuum evaporation presented a lower performance than UF in concentration of the
whey. Depending on the operating conditions ultrafiltration is able to concentrate whey 6-fold in 30 minutes,
while.

References

Almeida, K. E., Bonassi, I. A., Roça, R. O., (2001).Ciência Tecnol. Alim. 21(2).
AOAC. (1975). Official methods of analysis of the association of official analytical chemists. 12 ed. Washington. 2, 15.
Bassetti, F.J.; Peres, L. Petrus, J. C. C; Quadri, M. B.(2003) Aplicação de um modelo numérico na ultrafiltração de soro de
queijo. 4º Congresso Ibero-americano em Ciência e Tecnologia de Membranas. Florianópolis.
Behmer, M. L. A. (1986). Tecnologia do Leite. São Paulo: Nobel,.
Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the
principle of protein – dye binding. Anal. Biochem. 72, 248.
Chang Y.N., Huang, J.C., Lee, C.C., Shih, I.L. and Tzeng, Y.M. (2002), Use of response surface methodology to optimize
culture medium for production of lovastatin by Monascus rubber. Enzyme and Microbial Technology, 30, 889.
Chelini, D. B. P. (2002).Revista Milkbizz Tecnologia Temático Laticínios. 15.
Farro, A. P. C.; Viotto, L. A.. (2003). Redução do teor de gordura do soro de queijo pré-tratado por microfiltração. 4º
Congresso Iberi-americano em Ciência e Tecnologia de Membranas. Florianópolis. CITEM 2003.
Linden, G., Lorient, D. (1996) Bioquímica agroindustrial-Revalorización alimentaria de la producción agrícola.
Zaragosa(España): Acribia.
Pinheiro, A.I.R., Mosquim, M.C.A.V., Feres, P.A. (1993) Rev.Instit. Latic. Cândido Tostes, 48(287), 34.
8
 2nd Mercosur Congress on Chemical Engineering
th
4 Mercosur Congress on Process Systems Engineering

Sgarbieri V.C. (1996) Proteínas em alimentos proteicos, SP: Varela.

Silva, P.H.F., Pereira, D.B.C., Oliveira, L.L., Costa Jr., L.C.G. (1997). Físico-química do leite e derivados. Métodos
Analíticos. Ed. Oficina de Impressão Gráfica e Editora Ltda. Juiz de Fora, MG, 190p.
Tronco, V. M. Controle de Qualidade do Leite. Guaiba: Agropecuaria, 1996.
USDEC, U.S. Dairy Export Council. Disponível em :< http://www.usdec.org>. Accessed em 16 de janeiro de 2004
Veisseyre, R. (1998). Lactologia técnica: composicion, recogida, tratamiento y transformación de la leche. Zaragoza,
España: Acribia.

Acknowledgments

The authors acknowledge CAPES, Escola Agrotécnica Federal de Concórdia, URI-Campus de Erechim,
FAPERGS for financial support to this work; COCEL and TIROL for supplying cheese whey samples and
SENAI-Chapecó, SC for allowing the realization of experimental activities at its pilot UF plant.