Dr Buenaventura Guamis_UAB

1. Centre Especial de Recerca Planta de Tecnologia dels Aliments
AB Biotics
Felnuti
Ypsicon
CERPTA
www.cerpta.com

2. AB-BIOTICS
üProject Management R&D projects for companies.
üIn company R&D for development of functional
Main clients belong to the Food Functional and Food
ingredients, mainly probiotics for application in
supplements.
functional foods and food supplements.
üHistoric 5 years in operation and constant growth.
üCompany subsidiary dedicated to the development of üSubsidiary dedicated to developing a braking approach
advanced genetic tools for Medical Diagnosis (SNP, in oncology: membrane-lipid therapy.
CNV) üPartnerships with laboratories at Drug Discovery to
üThe company has appropriate human and technical ensure the inclusion of new molecules in the future.
resources to conducting genetic analysis.

4. YPSICON
Process-Plant Design, Installation &
Maintenance

5. CERPTA
The main research area at CERPTA
is the application of new
technologies, nanotechnologies
and technofunctionality for the
improvement of food safety, food
nutritional value and the design
and production of functional foods.

6. Centre Especial de Recerca Planta de Tecnologia dels Aliments
Fundamentos de alta
pressäo:aplicaçöes e equipamento
para a indústria de alimentos
www.cerpta.com

7. Origin of European Projects of HP
-La Grande-Motte Congress Montpellier- France
September of 1992

8. High Pressures
• Hydrostatic High
Pressures
• Dynamic High
Pressures (UHPH)

9. Hydrostatic High Pressures
Abismo Challenger 120 MPa
11 MPa = 9,869 atm = 10 bar = 10,197 kg/cm2

10. Hydrostatic High Pressures
11 MPa = 9,869 atm = 10 bar = 10,197 kg/cm2
Paul Regnard 1891

11. HISTORY
* Hite y col. (1899), Bacterial inactivation in foods
(milk, meat products and fruit juices).
* Giddings y col. (1929), Viruses inactivation.
* Timson y Short (1965), raw milk bacteria
inactivation.
* Inorganic materials (ceramics, metals, steels,
sintetics mat.) 70th.

12. HHP Biochemical and Biophysical.
Conclusions
• HP induces changes in the size, number, hydration,
composition and light-scattering properties of casein
micelles in HP-treated milk.
• Dairy whey hydrolysates obtained by pepsin and trypsin in
combination with HP treatment could be used as a source of
peptides in hypo-allergenic infant formulae.
• HP induces the association of whey proteins with casein
micelles which positively affects rennet properties of milk

13. HHP Biochemical and Biophysical.
Conclusions
• The amount of milk protein associated with the milk fat globules was
increased by HP treatment. HP-induced aggregation and denaturation of
agglutinins and lipoproteins are likely to have significant effects on HP-
induced changes in the creaming characteristics of milk
• High pressure treatment induces tertiary structural changes of BSA, but no
effect the secondary structure. We concluded that the pressure-induced
elimination of BSA allergenicity seemed to be related to the tertiary
structural change of BSA.
• The pressure-induced solubilisation of αS1- and αS2-caseins, essentially
located in the core of the micelles, suggests that high pressure
destabilised micelles including their internal structure.
• Unfolding of myosin and actin could be induced in extracted myofibrillar
protein with simultaneous treatment at 200 MPa and 40°C.

14. HHP Biochemical and Biophysical.
Conclusions
• Treatments of 500 MPa combined with storage at 4 ± 1 °C produce high
stability of lycopene when tomato puree was pressurized.
• In egg white proteins, pressure induces an increase in turbidity, surface
hydrophobicity, exposed SH content and susceptibility to enzymatic
hydrolysis, while it results in a decrease in protein solubility, total SH
content, denaturation enthalpy and trypsin inhibitory activity.
• Pressures of 300 MPa and above cause denaturation of β-conglycinin
(7S) and glycinin (11S) in soy milk. High pressure induces the formation
of tofu gels that have gel strength and a cross-linked network
microstructure.

15. HHP Biochemical and Biophysical.
Conclusions
• The solubility of dietary fibre in white cabbage can be
affected by high pressure temperature treatment, which may
be of importance when producing foods with specific health
effects.
• The effect of high-pressure processing (HPP) on cell wall
polysaccharides in berries was investigated.
• Compared to treatment at atmospheric pressure, pectic
polysaccharides were degraded to a larger extent when
HPP was used.

16. HHP Enzyme Conclusions
• Carrot PME is much more thermostable and pressure-stable in carrot
pieces than in carrot juice or purified form
• The catalytic activity of carrot PME was highly dependent on the
temperature and pressure applied. In model and food systems
(shredded carrots), optimal PME activity was registered at 50 °C in
combination with pressures of about 300–500 MPa
• Soybean whey proteins hydrolysed at high pressure could be used as
sources of peptides with low antigenicity when incorporated as food
ingredients.
• High pressure combined with suitable enzymatic activity could be a
useful tool for obtaining hydrolysates with low immunoreactivity to be
used in special foods (hypoallergenic foods).

17. HHP Enzyme Conclusions
• High pressure-induced inactivation of the indigenous milk enzymes
alkaline phosphatase (ALP), γ-glutamyltransferase (GGT) and
phosphohexoseisomerase (PHI) was studied in the pressure range 400-
800 MPa at temperatures between 5 and 40°C. With respect to pressure
stability the following ranking was observed: ALP>GGT>PHI.
• Combined thermal and high pressure inactivation of tomato
lipoxygenase occurs at pressures in the range of 100-650MPa combined
with temperatures from 10-60°C, and followed first-order kinetics. In the
high-temperature/low-pressure range, (T50°C and P300MPa) an
antagonistic effect is observed
• Individual and total carotenoids, and provitamin A carotenoids, were
significantly higher in HP tomato e than in the untreated and other
treated tomato es
• High pressure processing constitutes an effective technology to
inactivate the enzymes in fruit juices. Pressures higher than 400 MPa
can be combined with mild heat (<50 °C) to accelerate enzyme
inactivation.

18. Microorganisms

19. B. cereus ATCC 9139
Tratamientos AP Código
300 MPa / 15 min / 30ºC
300
400 MPa / 15 min / 30ºC
400
60 MPa / 210 min / 30ºC
G
60 MPa / 210 min / 30ºC + 300 MPa / 15 min / 30ºC G+300
60 MPa / 210 min / 30ºC + 400 MPa / 15 min / 30ºC G+400

20. B. cereus ATCC 9139
3
2,5
Reducción Log(No/N)
2
Sin aditivos
1,5 Nisina 1
Nisina 2
1 Lisozima
0,5
0
300 400 G G+300 G+400
Tratamientos AP

21. Evolución de recuentos de B. Cereus, 15 días a 8ºC.
Serie N2
7
C
Recuentos de B. Cereus
6 300
(log ufc/g)
400
5
G
4 G+300
G+400
3
0 5 10 15
días

22. Lethality in CIN of three strains of Y. enterocolitica inoculated
in model cheese and pressurized at 20ºC for 10 min at day 0.
Treatment Lethality (log No – log N)
Serotype
(MPa) Mean† CI‡
0 - -
300 ³3.36b ±0.41
O:1
400 ³3.36b ±0.41
500 ³3.36b ±0.41
Log (No/N)
0 - -
300 1.94c ±0.45
O:3
400 ³5.03a ±0.17
500 ³5.03a ±0.17
0 - -
300 3.48b ±0.17
O:8
400 ³4.28a ±0.40
500 ³4.28a ±0.40

23. Behaviour of Y. enterocolitica in model cheese after high
hydrostatic treatment (a) serotype O:1, (b) serotype O:3 and (c)
serotype O:8.
a
10
CIN Control
TALControl
9 CIN 300 MPa
TAL 300 MPa
8 CIN 400 and 500 MPa
TAL 400 and 500 MPa
7
log (CFU/g)
6
5
4
3
2
1
0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Time (Days) of storage at 8ºC

24. Behaviour of Y. enterocolitica in model cheese after high
hydrostatic treatment (a) serotype O:1, (b) serotype O:3 and (c)
serotype O:8.
b
10
CIN Control
TAL Control
9 CIN 300 MPa
TAL 300 MPa
8 CIN 400 and 500 MPa
TAL 400 and 500 MPa
7
log (CFU/g)
6
5
4
3
2
1
0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Time (Days) of storage at 8ºC

25. Behaviour of Y. enterocolitica in model cheese after high
hydrostatic treatment (a) serotype O:1, (b) serotype O:3 and (c)
serotype O:8.
c
10
CIN Control
TAL + CIN
9 CIN 300 MPa
TAL 300 MPa
8 CIN 400 and 500 MPa
TAL 400 and 500 MPa
7
log (CFU/g)
6
5
4
3
2
1
0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Time (Days) of storage at 8ºC

26. Lethality and counts in Sorbitol Mac Conkey agar of E. coli
O157:H7 inoculated in model cheese and pressurized at 12ºC
for 10 min at day 0.
Counts log (CFU/g)
Lethality (log No – log N)
Treatment
(MPa)
Mean† CI‡ Mean† CI‡
7.30
0 0.22 - -
300 3.71 0.88 3.59a 1.08
400 n.d - ³6.30b 0.22
500 n.d - ³6.30b 0.22

27. Behaviour of E. coli O157:H7 inoculated in model cheese after
HHP treatments at 300, 400 and 500 MPa.
10
Control TAL
Control Sorbitol
9 300 MPa TAL
300 MPa Sorbitol
8 400 MPa TAL
500 MPa TAL
7 400 and 500 MPa Sorbitol
Log (CFU/g)
6
5
4
3
2
1
0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Time (Days)

28. HHP Effects on Microorganisms
Conclusions
• For the inactivation of spores of B. subtilis and Clostridium sporogenes,, the
addition of nisin to the plating medium appeared to be synergistic in some
instances when combined with pressurization at elevated temperatures and
reduced pH. The adition of 0.1% sucrose laurate, may be dramatic synergistic
effects.
• HHP treatments at 5 degrees C induced more E. coli inactivations than those at
25 degrees C in liquid whole egg, from results of approximately 3 log reductions
of E. coli and over 5 log reductions of Pseudomonas and Paenibacillus, HHP
treatment of LWE (liquid whole egg) at 5 degrees C is regarded to be as
effective as conventional thermal pasteurization
• In skim milk suplemented with the lactoperoxidase-hydrogen peroxide-
thiocianate (LP) system considerable further inactivation of food borne bacteria
occurred in the first hours after pressure treatment.

29. HHP Effects on Microorganisms
Conclusions
• In minced chicken, vacuum packaging favour the growth of lactic acid
bacteria allowing the generation of desirable organisms as well as the
growth of spoilage indicators. High pressure treatment combined with
vacuum packaging and effective cold storage proved to be a very
effective tool for enhancing the microbial quality.
• Reduction of microorganisms increase when the rate of pressurization
and depressurization is increased.
• The high pressure pasteurisation processes are capable to inactivate
more than 5 log decades of the viable microorganisms present originally
in raw juice and product is free of coli-form bacteria, yeast, moulds and
salmonella during 30 days of storage at the chilled room temperature
conditions (temperature up to 5 °C). The high-pressure treated broccoli
juices are comparable in sulforaphane content and anti-mutagenic
activity with frozen version.

30. HHP Effects on Microorganisms
Conclusions
• The addition of nisin to cell suspensions after HP treatment, produce
irreversible effects in sublethal damages.
• Pressure inactivation of L. lactis is strongly temperature dependent,
baroprotection by sucrose occurs at any temperature but the
baroprotective effects of NaCl is temperature dependent.
• Whit combination of PEF and HHP processes, the non-treated spores
gradually turned into phase-dark spores and finally germinated and
changed into vegetative cells, while the spores subjected to PEF/HHP
treatment did not transfer to the phase-dark stage, meaning no
germination

31. HHP Effects on Microorganisms
Conclusions
• As expected, the rate of spore inactivation increased with
increasing pressure and temperature.
• A small fraction of the spore populations survives conditions
of up to 120°C and 1.4 GPa in isothermal treatments.
Because of this tailing and the fact that pressure-
temperature combinations stabilizing bacterial endospores
vary from strain to strain, food safety must be ensured in
case-by-case studies demonstrating inactivation or non
growth of C. botulinum with realistic contamination rates in
the respective pressurized food and equipment

32. Hydrostatic High Pressure Machinery
HP Cylinder
Heating/Cooling circuit
Product
Water exit Pressure intensifier
Decompression valve
Water entry
Low
pressure
pump

33. STANSTED-INMAPA

35. STANSTED

37. Industrial Machinery
Discontinuous Industrial Machine from EMBUTIDOS ESPUÑA (OLOT,
GIRONA)

38. AVURE

39. Hyperbaric

41. Semi-continuous Equipment

43. Treatment Cost of HHP
â Se debe considerar
– Costes amortización (relacionado con Pmáx del equipo)
– Mantenimiento y personal
– Consumo energía
Coste de inversión equipo AP para alimentos a 600 MPa
0,6-3 millones €
Tratamiento de 6000 L/h a 600 MPa, con un factor de eficiencia de
volumen del 50%, puede costar entre =0,0 5-0,25 €/L según sea la
productividad. El coste estimado está basado en una amortización a 5
años, con un 15% de interés, e incluye costes de tratamiento y
mantenimiento del equipo.

46. ESPUÑA (Spain): El pionero
• Primer producto cárnico HPP en el mundo.
• Lanzó en 1998 el primer jamón cocido loncheado
tratado por HPP.

47. MARTIKO (España)
Wave 6000/55
para productos de pato

48. ABRAHAM - ALEMANIA
Hiperbaric 150
Proscuitto alemán exportado a USA

50. KRAFT FOODS (Ameriqual) - USA
Wave 6000/300
Loncheado Natural

51. MAPLE LEAF (Canadá)
Wave 6000/300 para platos precocinados

52. Hiperbaric 55
MRM (España)

53. Moira Macs (Australia)
Hiperbaric 135

54. Vegetable products
• Higienización y aumento de la vida útil
• Conservación del color, sabor y vitaminas.
• Reducción de la actividad de la PPO
• Reducción de la retrogradación del arroz
Pa is Año Producto
Japan 1990 Mermeladas y salsas de frutas y verduras.
Japan 1994 Arroz pre-cocido e hipoalergénico.
USA 1997 Productos derivados del aguacate: guacamole
y salsas.
Italy 2001 Mermeladas de frutas.
USA 2002 Productos derivados del aguacate.
Mexico 2003 Productos derivados del aguacate..
Mexico 2003 Productos derivados del aguacate.
Mexico 2003 Productos derivados del aguacate.
USA 2003 Aros de cebolla.
Canada 2003 Productos a base de manzana: mermeladas y
salsas.
USA 2004 Tofu.
Spain 2005 Productos vegetales RTE.
USA 2006 Salsa de tomate.
USA 2007 Ensaladas
Australia 2008 Puré de frutas

55. Guacamole HPP

56. VERFRUCO (Mexico)
Hiperbaric 55
Guacamole y productos de aguacate

57. Hiperbaric 420
Sandridge (USA)
Para “wet salads” - PP

58. SANDRIDGE FOODS (USA)
Nuevo producto “premium” en 2011

59. SimplyFresco (USA)
Salsas
Stand Up Resealable Doy pack

60. CHIC FOODS
Hiperbaric 55 - PP

61. LEAHY ORCHARD: CANADA
• Puré de frutas
• Estable a temperatura
ambiente
http://www.hc-sc.gc.ca/fn-an/gmf-agm/appro/nf-
an108decdoc-eng.php

62. Drink and Juices
• Higienización y aumento de la vida útil
• Conservación del color, sabor y vitaminas.
• Reducción del amargor en el zumo de pomelo.
Compañia País Año Producto
Pokka Japan 1991 Zumo de uva.
Wakayama Japan 1992 Zumo de mandarina
Ulti France 1994 Zumos de cítricos.
Jumex Mexico 2000 Zumos de cítricos y smoothies
Ksun Lebanon 2001 Zumo s de frutas.
Lovitt Farms USA 2001 Zumo de manzana.
Frubaça Portugal 2001 Mezcla de zumos de manzana y cítricos.
Ata Italy 2001 Zumos de frutas y vegetales.
Avomex USA 2002 Zumos de naranja y limón y smoothies
Beskyd Czech Republic 2004 Zumos de remolacha, broccoli,
manzana y zanahoria.

63. Preshafruit – Donny Boy (Australia) con sus zumos HPP
ganó:
- Premio al mejor zumo
- Premio a mejor concepto de bebidas
At Beverage Innovation Awards ceremony, Drinktec 2009

64. Preshafruit – Donny Boy (Australia)

65. Coldpress (England) - PET

66. PET
Felixia-PET
FRUBAÇA (Portugal) –
PET/HDPE

67. Macè (Italy)
Hiperbaric 55 - Zumos & smoothies

68. EVOLUTION FRESH (USA)
Hiperbaric 420 en zumos - HDPE

69. Dairy Products
ü Destrucción de los microorganismos patógenos
ü Incremento de la productividad y reducción de costes.
ü Higienización y aumento de la vida útil
ü Estabilización de productos sin aditivos
ü Conservación de las emulsiones
País Año Producto
España 2007 Rellenos de sandwich con queso
EEUU 2007 Jerky cheese
N Zelanda 2009 Calostro

70. www.hiperbaric.com
RODILLA (España)
Rellenos para sándwich basado en queso o mayonesa y con ingredientes
Wave 6000/120
PET

71. NEW IMAGE Natural Health (NZ)
Hiperbaric 55
Bebida a base de calostro - HDPE

72. Fish Products
ü Extracción de la carne cruda de mariscos y crustáceos,
procesada pero no cocida.
ü Desarrollo de nuevos productos.
ü Incremento de la productividad y reducción de costes.
ü Higienización (inactivación de Vibrio)
ü Incremento de la vida útil.
País Año Producto
USA 1999 O stras
USA 2001 O stras
USA 2001 O stras
USA 2001 O stras
Canada 2004 Pescado
Canada 2004 Langosta
N. Zealand 2004 Mejillones en mitades.
Italy 2004 Bacalao desalado.
Spain 2004 Salmón, atún y merluza RTE
USA 2005 Langosta
Korea 2006 Mariscos
Canada 2006 Pescado
Japón 2007 Mariscos
Nota : letra azul = equipos de NC Hyperbaric

73. Supermarket in USA

74. High Pressure High Temperature (HPHT or PATS):
Vasija de 55 litros - 630 MPa - hasta +120ºC bajo presión
HIPERBARIC 55HT
ü Diseño horizontal:gradiente de temperatura reducido
ü Control independientre de temperatura del agua de presión
ü Sistema de carga y descarga automática
ü Control de temperatura de la vasija de presión
ü Sistema de calentado/enfriado

75. Ultra High Pressure Homogenization

76. 1900. First Homogenizers

77. 1902. Gaulin

78. Aims of Conventional Homogenizers
• Emulsion Stabilization
• Improved Taste
• Improved Texture
• Milk, Cream and Ice creams
• Pressures up to 50 MPa

79. 1980 High Pressure Homogenization is
Born (HPH)

80. HPH

81. HPH
Up to 150 MPa

82. HPH Conclusions
• More Stable Emulsions
• Improved Texture in Yogurts
• Microbial Counts Reduction

83. 1990 First Prototypes of Ultra High
Pressure Homogenization
200-250 MPa

84. 2003. First Valves and Prototypes of
UHPH with Double Intensifier Capable of
Working up to 350 MPa

85. Destrucción de microorganismos
HPH- UHPH
(a) (b)
TEM
Listeria monocytogenes inoculate in
PBS (0.01 M, pH 7.2)
(a) Untreated cells
(b, c, d) Treated Cells at 100, 200
and 300 MPa at 25ºC
(Vachon et al., 2002) (c) (d)

86. UHPH Effects on colloidal foods
Ø Particle size reduction Ø Microbial reduction
Ø Modification of colloidal Ø Enzymatic activity
structures reduction
Increasing physical
stability Food preservation
Without treatment 100 MPa
200 MPa 300 MPa
(Vachon et al., 2002)

87. UHPH and Milk and Dairy Products

88. Microbial inactivation by UHPH
Microbiological quality of goat milk for making
cheese treated by UHPH (log cfu/ml)
Treatment
Microbiota RA 100 200 300
Total bacteria 6.41 ± 1.38 a 5.54 ± 0.57 b 1.80 ± 0.52 c 1.39 ± 0.15 c
Psychrotrophic
bacteria 6.49 ± 1.36 a 5.62 ± 0.67 b 1.34 ± 0.21c 1.21 ± 0.36 c
Enterobacteriaceae 2.01± 0.85 a 0.96 ± 0.40 b ND ND
Lactobacilli 3.64 ± 0.38 a 3.12 ± 0.22 b ND ND
Lactococci 6.35 ± 1.46 a 5.41 ± 0.54 b 1.62 ± 0.56 c 0.95 ± 0.15 d
ND: not detected (under detection limit)
Quevedo et al. (2011)

89. Milk fat globule disruption
UHPH Conventional
homogenization
200 MPa 15-20 MPa
RA
X30.000
Particle size (µm)

90. Milk fat globule membrane
Transmission electron microscopy

91. Rennet coagulation aptitude
Quevedo et al. (2011)

92. Rennet coagulation
Confocal microscopy

93. Fresh cheese from UHPH milk
Cheesemaking
UH PA HP
Zamora et al. (2011)

94. Yogurt from UHPH milk
Yogurt making
Treatment Storage Time (days)
Firmness (N) 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
• Yogurts from UHPH-treated milk
are firmer than those from HT+SMP
HT+SMP 200 MPa
200 MPa
200 MPa
•UHPH gels are more
homogeneous, compact and less
porous than HT+SMP gels
•The fat fraction is much more
dipersed in UHPH gels and so, it is
completely incorporated into the
network
10µ 1010µ
10 µ
10µ
µ

95. Yogurt from UHPH milk
Yogurt making
ü Good aptitude of milk to acid coagulation without needing to add skim milk powder
ü Great firmness and water retention capacity during storage at cold temperatures
ü Low acidity and excellent sensory characteristics
Serra et al. (2007, 2008, 2009)

96. Effects of UHPH on milk
ü Similar microbial quality to high pasteurized milk (90ºC for 15 s) with a
treatment of 200-300 MPa and Ti = 30ºC or possibility to sterilization with 300
Mpa and Ti=75ºC compared to UHT milk.
ü Great physical stability against creaming during storage at cold temperatures
ü Whey protein denaturation and aggregation (UHPH at 300 MPa: 37%; PA at
90ºC for 15 s: 47%; b-Lg)
ü Slight reduction of micelle casein size
ü Slight mineral equilibrium alteration (transfer of soluble calcium and
phosphate to colloidal phase)
ü Fat globule disruption and composition of milk fat globule membrane altered
ü No lipolysis phenomena due to LPL inactivation by UHPH (200-300 MPa)
ü Good technologic aptitude to rennet and acid coagulations
Pereda et al. (2006, 2007, 2008, 2009) and Zamora et al. (2007)

97. UHPH and Vegetable Milks

98. Composition of vegetable milks
% w/w
Almond milk Soy milk
Dry matter 5.18±0.01 6.53±0.39
Protein 1.15±0.04 2.68±0.17
Fat 1.93±0.07 1.92±0.18
carbohydrates 1.99±0.02 1.35±0.05
ashes 0.11±0.04 0.18±0.05

99. Microbiological analysis
Almond milk Soy milk
Treatment Total counts* Spores* Total counts* Spores*
Control 4.4 ± 0.5 4.21 ± 0.07 4.21 ± 0.16 3.18 ± 0.06
UHT ND ND ND ND
Pasteurized 4.0 ± 0.18 3.60 ± 0.20 3.27 ± 0.17 2.27 ± 0.01
(1)
200, 55 3.21 ± 0.16 3.20 ± 0.03 3.06 ± 0.05 2.05 ± 0.07
200, 65 2.03 ± 0.07 1.83 ± 0.01 1.76 ± 0.17 1.02 ± 0.01
200, 75 ND ND ND ND
300, 55 ND ND ND ND
300, 65 ND ND ND ND
300,75 ND ND ND ND
(1) MPa, ºC: inlet temperature of UHPH-treated samples
* cfu/ml

100. Temperatures in UHPH
UHPH Inlet Outlet
treatments temperature T in the high temperature
of sample pressure of sample
(ºC) valve (ºC) (ºC)
(MPa)
200 55±0.5 105.7 ± 0.58 27.1 ± 1.0
300 55±0.5 128.3 ± 1.53 27.3 ± 1.1
200 65±0.5 111.7 ± 1.15 27.0 ± 1.0
300 65±0.5 130.7 ± 1.15 26.2 ± 0.8
200 75±0.5 117.0 ± 2.00 25.6 ± 2.7
300 75±0.5 135.7 ± 1.53 26.2 ± 2.2
Time less than 0.3 sec.

101. Sterility Study
Samples were taken aseptically into sterilized bottles and then
incubated at 30ºC for 20 days.
Treatment—Ti Tm-less 0.5sg Day 2 Day 7 Day 20
200 MPa 65ºC 111,7 coagulated
200 MPa 75ºC 130,7 coagulated
300 MPa 65ºC 117,0 - 3.97 ± 0.08 coagulated
300 MPa 75ºC 135,7 - - -
UHT (142ºC- 6 sec.) - - -

102. Sediment formation measured by turbiscan

103. Particle size analysis
Particle size (mm) of vegetable milks: Sauter
Particle size distribution in soymilk 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
0
eu T
75
l
20 5
30 5
20 5
75
65
20 d
ro
UH
6
5
5
e
0,
nt
riz
0,
0,
0,
0,
0,
30
Co
30
st
Pa

104. Stability index by centrifugation
(% w/w of sedimented particles)
10
9
8
7
6
5
Almond milk
4
Soymilk
3
2
1
0
Co UH Pa 20 20 20 30 30 30
nt T st 0, 0, 0, 0, 0, 0,
ro eu 55 65 75 55 65 75
l riz
ed

105. Oxidation (meq/L hydroperoxides)
0,45
0,4
0,35
0,3
0,25
0,2 Almond milk
0,15 Soymilk
0,1
0,05
0
T
l
75
ed
55
65
75
55
65
ro
UH
nt
0,
riz
0,
0,
0,
0,
0,
Co
30
eu
20
20
20
30
30
st
Pa

106. Reasons for applying UHPH to vegetable
milks
Ø Alternative to dairy milk
Ø Health benefits: fat fraction composition, rich in antioxidants,
balanced nutritional profile
Ø Processed by conventional heat treatments, specially UHT, which
implies heat damage
Ø Stability problems, specially sedimentation of solid particles and
creaming of fat globules

107. UHPH and Juices

108. TEMPERATURES DURING PROCESSING
Effect on temperature in apple juice during UHPH treatment at
different inlet temperature (Ti).
Temperatures (ºC)
Ti Pressures
T1 T2 To
(ºC) (MPa)
0 6.5 0.2 14.2 2.5 15.5 2.3
100 6.4 0.5 38.5 2.6 19.5 0.8
4
200 6.9 0.3 63.8 1.6 22.3 3.1
300 7.9 0.5 85.8 2.1 29.5 3.7
0 19.5 0.9 20.3 1.6 21.3 1.2
100 19.3 0.5 45.5 0.5 27.8 1.3
20
200 20.0 0.6 72.5 1.9 30.8 1.7
300 20.4 0.5 90.7 4.2 31.5 1.5
Mean Standard Deviation of three independent experiments (n=3) .
(T1) temperature just before the high-pressure valve, (T2) temperature
just after the high-pressure valve, (To) the outlet temperature. ~0.5 s

109. PHYSICOCHEMICAL ANALYSIS Apple
Juice
• BI • HMF
2,5
0,20 2,0
0,18
HMF mg/L)
1,5
0,16
(
0,14
1,0
0,12
Browning Index
0,10 0,5
0,08 20º C
0,0
4º C
0,06
Ctrl
Past
0,04 100 MPa
200 MPa
300 MPa
0,02
0,00
20º C
4º C
PA>>UHPH=R
Ctrl
Past
100 MPa
200 MPa 0,040
300 MPa
0,035
0,030
HMF ( g/L)
0,025
m
0,020
0,015
PA<R=100=200=300
0,010
0,005 20º C
0,000 4º C
Ctrl
100 MPa
200 MPa
300 MPa

110. Microbial analysis
8
.
Log (cfu/mL)
6
4
2
0
1 15 30 45 60
Days
Figure 1. Microbial population (log cfu/mL) of raw and treated apple juice during storage at 4 ºC.
The present data are the mean value of three experiments standard deviation (n=6). Where
A)Total Count B)Psichrotrophs C)Mould and Yeast D)Lactobacilli E)Enterobacteria and F)Faecal
Coliforms at differents treatments. Raw(¨), 100 MPa at Ti=4 ºC (¡), 100 MPa at Ti=20 ºC (•),
200M Pa at Ti=4 ºC ( ), 200 MPa at Ti=20 ºC (*), 300 MPa at Ti=4 ºC ( ), 300 MPa at Ti=20 ºC (+).

111. Enzymes
100 MPA 200 MPA 300 MPA PA
14.4/7.8 20.9/43.1
PME %
ND ND
%
PPO 30% 40% ND ND
0,20
0,18
0,16
0,14
Browning Index
0,12
0,10
0,08
0,06
0,04
0,02
20º C
0,00
4º C
Ctrl
Past
100 MPa
200 MPa
300 MPa

112. Vitamins
Treatment Ascorbic Acid Vit C Total b-Carotene
(mg/L) (mg/L) (mg/L)
R 0.22 0.03 a 6.77 1.10 b 11.38 0.59 d
100 at 4ºC 0.23 0.03 a 6.08 0.33 b 9.71 1.00 cd
200 at 4ºC 0.23 0.04 a 6.17 0.71 b 9.11 0.81 bc
300 at 4ºC 0.23 0.03 a 6.29 0.76 b 7.46 0.85 ab
100 at 20ºC 0.23 0.03a 6.56 0.96 b 7.75 1.00 ab
200 at 20ºC 0.23 0.02 a 6.47 0.88 b 7.48 0.09 ab
300 at 20ºC 0.24 0.03 a 6.49 0.88 b 7.67 0.35 ab
PA 0.23 0.04 a 0.76 0.14 a 6.82 0.43 a
R=UHPH=PA R=UHPH>>PA

113. ANTIOXIDANT CAPACITY
Antioxidan Capacity (ORAC) Antioxidant Capacity (TEAC)
Antioxidant Capacity (FRAP) Antioxidant Capacity (DPPH)
2000 Total phenolics (Folin-Ciocalteau) 60
-1)
Antioxidant Capacity ( mM TE)
Total phenolics (mg GAE L
c
1750
b b b b
a ab ab
1500 55
*
1250
.
1000 ** 50
750
500 45
250
0 40

114. Day: 0 Evolution Day: 15
Control 150 MPa 225 MPa 300 MPa
Control 150 MPa 225 MPa 300 MPa
Evolution Day: 30
Control
Evolution Day: 30
225 MPa
Control 150 MPa 225 MPa 300 MPa

115. It is possible to produce long life apple juice by
UHPH processing coupled with aseptic packaging.
UHPH treatment (300 MPa and Ti=5ºC) was selected
to preserve apple juice and was successfully coupled
to aseptic package technology. The UHPH-treated
juice could exhibe microbiological, nutritional,
physicochemical, biochemical and organoleptic
stability for about 21 months at 4 ºC (refrigeration
temperature).

116. UHPH and Orange Juice

117. Orange Juice
IT 10ºC/HT 30 sec IT 10ºC IT 20ºC
- PME activity as a
1,60
function of the applied
1,20 UHPH treatment.
UPE/ml
0,80
- For comparing also
0,40 control and thermal
0,00 pasteurization treatment
Control 100 MPa 200 MPa 300 MPa 90ºC/1min
data are shown.
Treatments

118. Orange Juice
- Counts of main
microbial groups
before and after
UHPH and thermal
pasteurization
treatments

119. Orange Juice
Antioxidant capacity
Ascorbic acid
Treatments *FRAP (mmol Trolox
(mg/100 ml)
equivalent/l)
Control (Fresh juice) 9,20 ± 1,26 a 51,06 ± 1,18 a
100 (IT 10 ºC/HT 30sec) 8,41 ± 1,23 a 50,48 ± 1,82 a
100 (IT 10 ºC) 7,88 ± 0,79 a 47,47 ± 0,40 c
-Antioxidant power and
100 (IT 20 ºC) 8,23 ± 2,44 a 45,06 ± 0,35 de
total content of ascorbic
200 (IT 10 ºC/HT 30sec) 8,42 ± 1,35 a 48,73 ± 0,70 b
acid in the samples before
200 (IT 10 ºC) 7,78 ± 1,10 a 46,58 ± 0,35 c
and after UHPH and
200 (IT 20 ºC) 7,91 ± 1,71 a 47,62 ± 0,45 c
thermal pasteurization
treatments.
300 (IT 10 ºC/HT 30sec) 7,78 ± 0,99 a 46,87 ± 1,43 c
300 (IT 10 ºC) 7,60 ± 0,88 a 44,21 ± 1,67 e
300 (IT 20 ºC) 8,06 ± 1,73 a 45,51 ± 0,42 d
Pasteurized (90ºC/1min) 8,66 ± 1,15 a 45,87 ± 0,42 de

120. Submicroencapsulation

121. Submicroencapsulation
UHPH up to
400 MPa

122. Results
Casein (2,5%) + β-Carotene (0,01%)
Control 300 MPa

123. Turbiscan
UNSTABLE DISPERSIONS
STABLE DISPERSIONS

124. TEM Casein submicrocapsules

125. UHPH System 15 Prototype.

126. UHPH System 100 Prototype.

127. Acknowledgments:
Ultra high pressure homogenization
UAB team:
Buenaventura Guamis Martín Buffa
Victoria Ferragut Ramón Gervilla
Toni Trujillo Jordi Saldo
Artur Xavier Roig Maria del Mar Serra
Manoli Fernández Ana Zamora
Joan Miquel Quevedo Angela Suarez
Tomás López Roger Escriu

128. Acknowledgments:
Funentech EU Project
Stansted Fluid Powert
ABBiotics
Nectina
UAB
UMPII
Max Rubner-Institut (MRI)
Thank you for your attention