Principales aplicaciones de la UHPH

• Leche y derivados
– Leche de consumo
– Yogurt
– Queso fresco
– Queso madurado

• Licuados vegetales
– Bebidas
– Productos fermentados

• Zumos
 LECHE
CARACTERÍSTICAS MICROBIOLÓGICAS,
FISICOQUÍMICAS Y ENZIMÁTICAS.
ESTUDIO DE LA VIDA ÚTIL
 UHPH: Described effects on some milk
constituents and enzymes
Constituent Described effect References
Thiebaud et al., 2003; Hayes et al.,
Decrease in average size
Fat globules and formation of clusters
2003; Pereda et al., 2007; Zamora, et
al. 2007
Hayes et al., 2003; Datta et al., 2005;
Denaturation, although big
Whey proteins variability in the results
Sandra et al., 2005; Bouaouina et al.,
2006; Zamora et al., 2007
Decrease in average size Sandra et al., 2005; Hayes et al.,
Casein micelles 2003

Inactivation at 200 MPa at

Lactoperoxidase ≥ 40 ºC
Datta et al., 2005; Pereda et al., 2007

Hayes et al., 2003; Datta et al., 2005;

Plasmin Inactivation
Pereda et al., 2008

Alkaline Inactivation at 200 and 300 Hayes et al., 2003; Picart et al., 2006;
phosphatase MPa at 30 ºC and 40 ºC Pereda et al., 2007

Lipoprotein Inactivation at outlet

Datta et al., 2005
lipase temperature >70 ºC
 UHPH: Described effects on some
functional properties of milk

Properties Described effects References

Lower curd firmning rate,

Rennet coagulation higher curd firmness and Hayes et al., 2003; Zamora
properties higher curd yield in et al., 2007
treatments at one stage

Stabilizing properties of Better stabilizing and

Bouaouina et al., 2006
whey proteins increased foaming ability

Cheddar cheese: Higher

cheese yield and more
Guerzoni et al. 1999; Kheadr
Cheese-making compact structure
et al., 2002; Lanciotti et al.,
properties Ripenned goat cheese: 2004
different proteolytic and
lipolytic profiles
 Modificación de la temperatura de la leche durante los
tratamientos UHPH

Temperatura de Presión T1 (ºC) T2 (ºC)

entrada (Ti) homogenización
(MPa)
100 43,0 + 1,0 64,7 + 1,5

40ºC
200 46,7 + 0,6 84,0 + 1,0

300 50,0 + 1,0 103,0 + 1,0

200 39,0 + 0,0 78,7 + 1,2

30ºC
300 42,7 + 0,6 97,3 + 0,6
 Leche: Efectos del tratamiento

Psychotrophic
Total counts* Lactococci*
Treatment bacteria*
Raw 5.1 ± 0.5 4.9 ± 0.3 5.3 ± 0.3
Pasteurised 0.4 ± 0.7 0.5 ± 0.8 ND
200 MPa 30°C 1.3 ± 0.1 1.0 ± 0.4 0.2 ± 0.4
200 MPa 40°C 0.9 ± 0.4 0.9 ± 0.4 0.3 ± 0.5
300 MPa 30°C 1.0 ± 0.5 1.0 ± 0.4 0.1 ± 0.2
300 MPa 40°C 1.0 ± 0.6 0.8 ± 0.4 0.3 ± 0.5
* Log cfu/ml

PhD Thesis: Pereda, 2008

 Leche: Tamaño de partícula

Milk fat globules

Conventional
homogenisation

UHPH
200 MPa Raw
Volum (%)

Particle size (mm)

PhD Thesis: Zamora, 2009

 Leche: evolución del pH durante
el almacenamiento

Evolución del pH

6,78
.

6,76

6,74
pH (unidades)

6,72

6,70

6,68

6,66
0 3 6 9 12 15 18 21

Tiempo de almacenamiento ( días)

200 30ºC 200 40ºC 300 30ºC 300 40ºC PAST

 Leche: enzimas

Tratamiento Lactoperoxidasa (U/ml) Fosfatasa alcalina (U/ml)

Leche cruda 1,34 a ± 0,03 35% 0,88 a ± 0,01

actividad
residual
Pasteurizada Nd c Nd b
0,46 b ± 0,01
200 MPa, 30ºC 0.46 b ± 0.01 Nd b

200 MPa, 40ºC 0,01 c ± 0,00 Nd b

300 MPa, 30ºC Nd c Nd b

300 MPa, 40ºC Nd c Nd b

Leche: marcadores térmicos

Furosine

Whey proteins

Lactulose

PhD Thesis: Pereda, 2008

 LECHE

CARACTERÍSTICAS DE COAGULACIÓN ENZIMÁTICA

QUESO FRESCO
Coagulación enzimática:curvas de coagulación
Queso fresco
Producción
Queso fresco
 Queso fresco: microbiología

• No pathogens

• Almost no coliforms

• No lactobacilli

• Very low counts of

yeasts & moulds
Coagulación enzimática:proteínas del suero
Coagulación enzimática: rendimiento bruto, humedad de
la cuajada y rendimiento de sólidos totales
 Queso fresco: microstructura

Confocal laser scanning microscopy

UHPH
300 MPa 30°C Water typology

Past Homo-Past

PhD Thesis: Zamora, 2009

 LECHE
CARACTERÍSTICAS DE COAGULACIÓN ÁCIDA
YOGUR
 Coagulación ácida
10 ml inoculated milk

80 4,5
70 4
60 D1 3,5
Signal (mA)

Signal (mA)
AR 3
50
2,5
40
2
30 D0
1,5
20 1
10 CT 0,5
0 0
30 60 90 120 150 180 210 240
Time (min)

• Coagulation time
• Aggregation rate
• Network density
 Coagulación ácida: velocidad de agregación

Tratamiento Velocidad de agregación

Control 0,62±0,01d
Control+ES 0,89±0,01a,b
30ºC 40ºC
100 MPa 0,88 ±0,13a,b 1,0 ±0,04a

200 MPa 0,96 ±0,1a 0,89 ±0,08a,b

300 MPa 0,97 ±0,03a 0,99 ±0,03a

 Coagulación ácida: densidad de la red (firmeza)

Calculado de las curvas de coagulación (43ºC)

Firmeza:
- Cristalización de la grasa Fenomenos a
- Interacción puentes de hidrógeno temperaturas bajas

30ºC 40ºC

50
Signal (mA)

40
30
20
10
0
100 130 200 230 300 330 HT HT+SMP
Treatm ent (MPa)
 Coagulación ácida: capacidad de retención de agua
yogures

Whey expelled by centrifugation

30ºC 40ºC

80 a
a a a
% whey expelled (w/w)

70 a a
b c c
60 c c c c
c
50
40
30
20
10
0
100 130 200 230 300 330 HT HT+SMP
Treatm ent (MPa)
90ºC, 90s
15 MPa

Protein-protein interactions 3% SMP added

Fat-protein interactions: fat Whey proteins denaturation by heat
globules behave like casein micelles
 Coagulación ácida: textura
yogures

30ºC 40ºC

2,5 a
b b
bc bc
c c c
2 c
Firmness (N)

1,5 d
e e
1 e e

0,5

0
100 130 200 230 300 330 HT HT+SMP

Treatment (MPa)

3% SMP added
Protein-protein interactions
Whey proteins denaturation by heat
Fat-protein interactions
 Coagulación ácida: microstructura
yogures
HT+SMP 200 MPa 300 MPa

10μ 1010
10μ
μμ
10μ 1010μ
10μ
μ

Branched Denser microstructure

microstructure
Low amount of DWP interacting with caseins.
Partial micelle disintegration leads to a higher
The presence of DWP on the casein
solvation of these proteins, but also to the
surface hinders the formation of large
solubilization of CCP
casein aggregates.

WP-CN linkages CN-CN linkages (casein fusion)

Coagulación ácida: evolución de la viscoelasticidad de
yogures durante el almacenamiento en refrigeración

G’ values were higher in gels from HT+SMP:

– Higher number of bonds because of 3% more of total solids
– Linkages between WP and CN are via disulphide bonds, which are
stronger than CN-CN unions present in UHPH gels (hydrophobic and
electrostatic interactions)

G’ values slightly increased and tan (δ) values decreased with time due to
particle rearrangements that consisted on the formation of more bonds per
junction of the network.
0,26
0,25

0,24
tan

0,23

0,22

0,21
HT+SMP 200 300

Treatm ent
1 7 14 21 28

Favoured by cold temperature and overacidification which intensify

intreactions between CN by means of CCP solubilization.
 Coagulación ácida: sinéresis yogures durante el
almacenamiento
a a
a a
60 a
b
b
% Whey expelled 50 c b b
c b
40 c c c
30
20
10
0
1 7 14 21 28
Storage time (days)
HT+SMP 200 300

In UHPH samples:
1- Increased nº of micellar particles
2- High solvation of these particles
3- High dispersion of the fat fraction

In agreement with G’ and tan(δ) values, the increase in the number of

junctions resulted in structures more capable of supporting centrifugal forces.
 Coagulación ácida: evolución de la textura de yogures
durante le almacenamiento

Yogurts from UHPH-treated milk were firmer than those from HT+SMP

Treatment Storage Time (days)

1 7 14 21 28
HT+SMP 1.42a 1.42a 1.43a 1.37a 1.42a
200 MPa 1.83b 1.79b 1.81b 1.86b 1.87b
300 MPa 1.99c 2.13c 2.27c 2.33c 2.23c

• UHPH gels are more homogeneous, compact and less porus than HT+SMP
gels

• The fat fraction is much more dipersed in UHPH gels and so, is completely
incorporated into the network

Texture test is not sensitive enough to reflect changes that occur at

particle-particle level
 LICUADOS VEGETALES
BEBIDAS
PRODUCTOS FERMENTADOS
 Licuados vegetales: Microbiología
Efectos del tratamiento

Almond drink Soy drink

Treatment Total counts* Spores* Total counts* Spores*
Control 4.4 ± 0.5 4.2 ± 0.1 4.2 ± 0.2 3.2 ± 0.1
UHT ND ND ND ND
Pasteurised 4.0 ± 0.2 3.6 ± 0.2 3.3 ± 0.2 2.3 ± 0.1
200 MPa 55°C 3.2 ± 0.2 3.2 ± 0.1 3.1 ± 0.1 2.1 ± 0.1
200 MPa 65°C 2.1 ± 0.1 1.8 ± 0.1 1.8 ± 0.2 1.0 ± 0.1
200 MPa 75°C ND ND ND ND
300 MPa 55°C ND ND ND ND
300 MPa 65°C ND ND ND ND
300 MPa 75°C ND ND ND ND
* Log cfu/ml
PhD Thesis: Cruz, 2008
 Licuados vegetales:
Microbiología durante el almacenamiento

6
4.2
Log counts (cfu/mL)

5
3.2
4

2
0.3
1

0
0 7 14 21 28

Days

6,9

6,8
Figure 10. Pasteurized soymilk (-♦-), 200 MPa,
55ºC (-■-) and 200 MPa, 75ºC soymilks (-▲-).
pH

6,7

6,6

6,5
0 7 14 21 28

Days
 Licuados vegetales: bebidas

Particle size (mm) of vegetable drinks: Sauter

Particle size distribution in soy drink Diameter (d3,2)

1,4
300 MPa
1,2

1
200 MPa
0,8

Pasteurized 0,6 Almond milk

UHT Soymilk
0,4

Raw 0,2

eu T

75
l

20 5

20 5
75

30 5
65
20 d
ro
UH

5
e

0,
nt

riz

0,

0,

0,

0,

0,
30
Co

30
st
Pa
 Licuados vegetales: bebidas

Raw UHT UHPH

PhD Thesis: Cruz, 2008

 Licuados vegetales:
hidrofobicidad licuado de soja

Caracterización de la superficie hidrófoba de la proteína

Figure 16. Protein surface hydrophobicity of BP (―), UHT-treated soymilk (―) and UHPH-treated soymilk at 300 MPa 80ºC (―).
 Licuados vegetales:
Estabilidad coloidal licuados de soja
Cruda

18%

Pasteurizada

13%

200 MPa, 75ºC

7.6%
Micrografías TEM de los licuados de soja PB, UHT y UHPH. (gg) glóbulos de grasa, (ap) agregados
de proteína, (agg) agregados de glóbulos de grasa, (apg) agregados de proteínas-glóbulos de
grasa, (p) proteína (q) agregados de inclusión de microgotas de grasa en proteína.
 Licuados vegetales:
Oxidación licuado de soja

Oxidación lipídica: Actividad lipoxigenase no detectada.

1,4
i
1,2
(meq peroxide/L sample)

1
Lipid hydroperoxides

h
hj j
0,8 k
k
k
0,6
b l
cd d de
0,4 ce c ce
g a
a
0,2

0
BP Past UHT 200MPa 200MPa 200MPa 300MPa 300MPa 300MPa
55ºC 65ºC 75ºC 55ºC 65ºC 75ºC

Figure 6. Hydroperoxides values (mean ± SD) of BP, heat treated (pasteurized and UHT) and
UHPH soymilks at day 1 ( ) and day 15 ( ).
a-e Different letters above bars of day 1 indicate that samples are significantly different (P < 0.05).
g-l Different letters above bars of day 15 indicate that samples are significantly different (P < 0.05).
 Licuados vegetales:
Inhibidores de tripsina

9.25 g of TI/L

3
a 80%
2,5

b 60.8% b
2 b b b b
g of TI/L of sample

1,5
c
1

0,5

0
BP Past UHT 200MPa 200MPa 200MPa 300MPa 300MPa 300MPa
55ºC 65ºC 75ºC 55ºC 65ºC 75ºC

Figure 7. Trypsin inhibitor values (mean ± SD) of BP, heat-treated and UHPH soymilks.
a-c Different letters above bars of each treatment indicate that samples are significantly different (P < 0.05).
 Licuados vegetales: esterilización
300 MPa, 75 ºC

Análisis microbiológico: Recuento total: 3.18 ± 0.73 log cfu/mL

Esporas: 2.18 ± 0.53 log cfu/mL
Producto base
Enterobactérias: 0.49 ± 0.19 log cfu/mL
Bacillus cereus: 2.29 ± 0.78 log cfu/mL

6,9

6,8
pH

6,7

6,6
0 30 60 90 120 150 180

Days

Figure 16. pH measurements of UHPH soymilk (-♦-) and UHT soymilk (-■-).
 Licuados vegetales: esterilización
Evolución durante el almacenamiento

Tamaño de partícula Sedimentación

8
Parameters Day UHT UHPH b b
b b
7
D50A 1 0.45 ± 0.03xa 0.17 ± 0.01ya ab
ab
ab
6
0.47 ± 0.06xa 0.18 ± 0.01yb

% Solids sedimentation
90 a
180 0.32 ± 0.06xb 0.17 ± 0.01ya 5

d3,2B 1 0.36 ± 0.02xa 0.23 ± 0.19xa 4

a a
90 0.35 ± 0.02xa 0.17 ± 0.01ya 3 ab a
ab ab ab b
180 0.26 ± 0.05xb 0.16 ± 0.01ya 2
d4,3 C 1 14.03 ± 0.72xa 6.14 ± 1.64ya 1
90 15.46 ± 2.03xab 5.07 ± 1.15ya
0
180 16.57 ± 1.94xb 6.38 ± 0.83ya 1 20 40 60 90 120 150 180
a-b Days
Different superscripts in the same column are significantly different (P < 0.05).
x-yDifferent superscripts in the same row are significantly different (P < 0.05).
A Mean values ± SD (µm) of diameter below which 50% of the volume of particles are Figure 17. Solids sedimentation of UHT-treated soymilk
found. ( ) and UHPH-treated soymilk ( ).
B Mean values ± SD (µm) of average diameter (surface weighted mean diameter). a-b Different superscript for the same sample are
C Mean values ± SD (µm) of average diameter (volume weighted mean diameter). significantly different (P < 0.05).
 Licuados vegetales: productos fermentados

Streptococcus thermophilus Lactobacillus delbrueckii spp bulgaricus

10,00 10,00
9,00 9,00

Log (UFC/ml) .
Log (UFC/ml) .

8,00 8,00
7,00 7,00
6,00 6,00
5,00 5,00
4,00 4,00
3,00 3,00
1 7 14 21 28 1 7 14 21 28

Tiempo (días) Tiempo (días)

95ºC 15 min 200 MPa 50ºC 300 MPa 50ºC 300 MPa 50ºC, 15 s 95ºC 15 min 200 MPa 50ºC 300 MPa 50ºC 300 MPa 50ºC, 15 s

Supervivencia de cultivos iniciadores en el yogurt de soja elaborados con licuado de soja tratado a 95ºC, 15 min y por UHPH.
Licuados vegetales: productos fermentados

Raw Pasteurised 200 MPa 300 MPa

Día1 95ºC 15 min Día 28

10 μm 10 μm

200 MPa 50ºC

Micrografías por microscopía confocal de la matriz proteica

10 μm 10 μm de yogures de soja elaborado con licuado de soja tratados a
300 MPa 50ºC 95ºC, 15 y por UHPH del día 1 y 28 de almacenado.

10 μm 10 μm

300 MPa 50ºC/15 seg

10 μm 10 μm
 Licuados vegetales: productos fermentados
Evolución de la viscoelasticidad

Dia1 Día 28
3000
3000
2500 2500
G' G'

G´G" (Pa) .
G´G" (Pa) .

2000 2000
1500 1500
1000 1000

500 G" 500 G"

0 0
0,1 0,15 0,22 0,32 0,46 0,68 1 0,1 0,15 0,22 0,32 0,46 0,68 1
f(Hz)
f(Hz)
95ºC 15´ 200 MPa 50ºC 300 MPa 50ºC 300 MPa 50ºC/15 s 95ºC 15´ 200 M Pa 50ºC 300 M Pa 50ºC 300 M Pa 50ºC/15 s

Parámetros G' G" (Pa) de yogures de soja elaborados con licuados de soja tratados a 95ºC, 15 min y por UHPH
del día 1 y 28 de almacenamiento.
ZUMOS FRUTAS
Enzymes and bioactive compounds
Orange juice:
Pectin methyl esterase (PME)
Biocompounds: carotenoids, flavonoids,
polyphenols & antioxidant activity
Apple juice:
Polyphenol oxidase (PPO) & Pectin methyl
esterase (PME)
Hydroxymethylfurfural (HMF)
Ascorbic acid and vitamin C
 Zumos: naranja
actividad pectinmetilesterasa

≤0.7 seconds
PME Remaining activity (%)

100 a a a 30 seconds
b
80

60

40

20 c F P
c c c 100 200 300
c c c c c c
0
100/10
10ºC 100/20
20ºC 200/10
10ºC 200/20
20ºC 300/10
10ºC 300/20
20ºC 1min 2min
100 MPa 200 MPa 300 MPa 90ºC
Treatments

PhD Thesis: Velázquez, 2011

 Zumos: naranja
compuestos bioactivos

Fresh 200 MPa 300 MPa Pasteurised

Carotenoids
147.4 ± 54.0a 136.3 ± 67.5ab 106.8 ± 66.5ab 87.3 ± 33.6b
(mg/100mL)
Flavonoids
12.3 ± 2.1a 21.0 ± 7.4b 23.0 ± 8.3b 14.1 ± 3.8a
(mg/100mL)
Polyphenols
77.1 ± 12.2a 75.9 ± 9.7a 72.0 ± 10.3a 62.4 ± 17.2b
(mg/100mL)
FRAP (mM) 9.7 ± 1.9a 8.7 ± 1.8b 8.1 ± 1.5b 8.7 ± 1.4b
TEAC (mM) 6.1 ± 0.9a 6.4 ± 1.0a 6.1 ± 0.9a 5.6 ± 0.8b

PhD Thesis: Velázquez, 2011

 Zumos: manzana
actividad enzimática y daño térmico

Hydroxymethylfurfural (HMF)
Polyphenol oxidase (PPO) &
Pectin methyl esterase (PME)

Treatment PPO PME

200 MPa 40% 20.9/43.1%
300 MPa ND ND
Pasteurised ND ND

PhD Thesis: Suárez, 2011

 Zumos: manzana
Componentes bioactivos

Ascorbic Acid Total Vit C b-Carotene

(mg/L) (mg/L) (mg/L)
Treatment
Raw 0.22 ± 0.03 a 6.77 ± 1.10 b 11.38 ± 0.59 d
200 at 4°C 0.23 ± 0.04 a 6.17 ± 0.71 b 9.11 ± 0.81 bc
300 at 4°C 0.23 ± 0.03 a 6.29 ± 0.76 b 7.46 ± 0.85 ab
200 at 20°C 0.23 ± 0.02 a 6.47 ± 0.88 b 7.48 ± 0.09 ab
300 at 20°C 0.24 ± 0.03 a 6.49 ± 0.88 b 7.67 ± 0.35 ab
Pasteurised 0.23 ± 0.04 a 0.76 ± 0.14 a 6.82 ± 0.43 a

PhD Thesis: Suárez, 2011

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