TRENDS IN HIGH PRESSURE PROCESSING OF FOODS OOD …allnaturalfreshness.com/wp-content/uploads/2014/02/Trends-in-HPP.pdf · FOOD QUALITY AND BIOACTIVE COMPONENTS Shirani Gamlath1 and

In: New Topics in Food Engineering ISBN: 978-1-61209-599-8 Editor: Mariann A. Comeau © 2011 Nova Science Publishers, Inc.

Chapter 6

TRENDS IN HIGH PRESSURE PROCESSING OF FOODS: FOOD QUALITY AND BIOACTIVE COMPONENTS

Shirani Gamlath1 and Lara Wakeling2

1 School of Exercise & Nutrition Sciences, Faculty of Health, Medicine, Nursing & Behavioural Sciences, Deakin University, Victoria, 3125, Australia

2 School of Science & Engineering, University of Ballarat, Ballarat, Victoria, 3353, Australia

ABSTRACT

High pressure processing (HPP) is a non-thermal food processing technology that offers great potential for the processing of a wide range of food products. Application of HPP can inactivate micro-organisms, affect food-related enzymes and modify structures with minimal changes to nutritional and sensory quality aspects of foods. The effects of high pressure on the inactivation of micro-organisms in food have been thoroughly reviewed. Recent research on HPP has mainly focused on fruits and vegetables with an emphasis on food quality and bioactive components. This chapter highlights the current trends in HPP research and provides a summary of the available findings on the effect of HPP on chemical, nutritional and bioactive components and health related properties of a wider range of commodities. Strategies to maintain the quality attributes and health related components in HPP foods and identification of the gaps for future research in HPP are also discussed.

INTRODUCTION High pressure processing (HPP) has gained much attention in recent years due to a

number of advantages: the retention of fresh taste and texture in products such as fruit and vegetable juices, shell fish, sauces and guacamole; an increase in microbiological safety and shelf-life by inactivation of pathogens, spoilage organisms and some quality related enzymes; production of novel products by modifying the existing food structures; low energy

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Shirani Gamlath and Lara Wakeling 122

consumption and labour requirements, and uniform isostatic pressure distribution throughout the product, irrespective of size and geometry (Patterson, 2006; Rastogi et al., 2007).

A number of publications have demonstrated the application of HPP in relation to microbiological safety and keeping quality of foods, with optimal pressure and exposure times to validate the process conditions for fruits, vegetable, dairy and meat products. Some quality attributes, particularly the effect on quality related enzymes in food matrices and buffer systems, have also been extensively studied.

There is a significant commercial interest in both the local and global markets for new food products particularly fresh-like food products with extended shelf life and high quality. The interest of consumers in the consumption of fresh fruit and vegetables with high antioxidant levels has also become a current trend. High pressure processing has attracted local and global interest due to its mild effect on food products, while maintaining high nutritional value and fresh like properties compared to thermal processing. However, the existing knowledge on pressure stability of nutritional and bioactive components in HPP foods and the optimum conditions to retain the beneficial properties of foods are limited.

This chapter aims to explore the current literature around the effect of HPP on nutritional, chemical and bioactive components and health related properties of a wider range of commodities with an emphasis on identifying suitable HPP regimes to retain the nutritional and healthful properties of foods.

EFFECT OF HPP ON FOOD QUALITY It is now well known that HPP can be used to inactivate micro-organisms, while

maintaining organoleptic and nutritional qualities of the food. Much of this quality work has focussed on colour and texture in food, as these are the properties that consumers base their food choices on. The effect of HPP on colour and texture has been extensively covered in fruit and vegetables over many years, with comprehensive reviews available by Guerrero-Beltran et al., (2005), Rostagi et al., (2007) and Norton and Sun (2008).

Most texture related investigations have focussed on the effect of high pressure on enzymes such as pectin methylesterase and polygaluctonase, although interest in assessing texture by instrumental means is now increasing (Norton and Sun, 2008, Perera et al., 2010). In muscle foods increased pressures are thought to result in changes to the muscle proteins, resulting in changes to functional properties and therefore to texture. This was found to be the case in fish where pressures ≥450 MPa resulted in higher hardness, gumminess and chewiness (Yagiz et al., 2007).

Colour has been assessed via either instrumental techniques or individual pigments, such as the carotenoids, have been assessed. Table 1 summarises some very recent findings in this area. There is a decrease in redness in all muscle foods after HPP (Andres et al., 2006, Cava et al., 2009) with a corresponding increase in lightness of fish, often associated with a cooked appearance (Ramirez-Suraz and Morrissey, 2006, Yagiz et al., 2007). A decrease in the lightness of HPP cow’s milk and ewe’s milk has also been observed (San Martin-Gonzalez et al., 2006). It has been suggested that the changes in the lightness of these animal products are due to protein structural changes that occur after HPP.

Table 1. The effect of HPP on colour- examples from recent studies

Commodity Method used HPP conditions* Compared to the experimental control

Reference

Iberian ham Iberian loin

Colour 200 & 400 MPa/20 °C/15 min 200 & 300 MPa/15-30 min 200 & 300 MPa/15-30 min

Lower initial redness (a*) then stable Lightness ↑ with storage (39 days at 5 °C) ↓ redness; lightness and yellowness unchanged. Similar effect after 90 days storage (4 °C) ↓ redness; lightness and yellowness unchanged. Similar effect after 90 days storage (4 °C), except yellowness ↑

Andres et al., 2006 Cava et al., 2009 Cava et al., 2009

Rainbow trout (whole muscle)

Colour (up to 6 days)

150, 300, 450, 600 MPa/RT/15 min

↓ redness (a*) >150 MPa cooked appearance (↑ lightness)

Yagiz et al., 2007

Mahi Mahi (whole muscle)

Colour (up to 6 days)

150, 300, 450, 600 MPa/RT/15 min

↓ redness (a*) Slight ↑ in lightness & yellowness with pressure.

Yagiz et al., 2007

Albacore tuna (minced muscle)

Colour 275 & 310 MPa /RT/2, 4, 6 min

Lighter and whiter with ‘cooked appearance’; (L* and b* ↑; a* ↓)

Ramirez-Suraz and Morrissey, 2006

Cows and Ewe’s milk ∆E ↓ in lightness San Martin-Gonzalez et al., 2006

Melon

Colour 600 MPa/RT/10 min ↑ in lightness, redness and yellowness

Wolbang et al., 2008

Table 1. (Continued)

Commodity Method used HPP conditions* Compared to the experimental control

Reference

Apple – Granny Smith Pink Lady

∆E 600 MPa/ 22 °C/1-5 min (medium: pineapple juice)

↓ at HPP for 3 & 5 min + 25 & 50 % pineapple juice ↓ at HPP for 1 min + 25 % pineapple juice

Perera et al., 2010

Blackberry puree Strawberry puree

Colour & ∆E 400, 500 & 600 MPa/10-30 °C/15min

Minor changes. Redness ↓; 96 % retention in strawberries at 600 MPa

Patras et al., 2009a

Tomato puree Carrot puree

Colour 400-600 MPa/20 °C/15 min

Redness and colour intensity retained. Lightness ↑ in carrots but ↓ in tomatoes

Patras et al., 2009b

Tomato juice and Carrot juice

∆E 250 MPa/35 °C/15 min

Minor change with HPP. Change ∆E≤15 units after storage for 30 days (4 & 25 °C)

Dede et al., 2007

* RT = Room temperature

Trends in High Pressure Processing of Foods: Food Quality … 125

Minimal changes occur to the colour of HPP fruits (Dede et al., 2007; Patras et al., 2009a,b; Perera et al., 2010), although changes may occur in individual pigments. Further information about the affect of HPP on individual pigments is given in the section on bioactive compounds.

It is generally accepted that HPP does not alter the flavour of food. However a recent application of HPP regarding flavour has been the combined use of enzyme immobilisation and HPP to de-bitter grapefruit juice (Ferreira et al., 2008). A pressure of 160 MPa/37 °C/20 min resulted in a reduction in the naringin content and therefore a reduction in bitterness.

EFFECT OF HPP ON CHEMICAL AND NUTRITIONAL COMPONENTS IN FOOD

Macronutrients

Some recent studies highlighting the effect of HPP on macronutrients in food are shown

in Table 2. Generally the total protein and total lipid contents are not affected by HPP. Lipid tends to be of greatest interest in terms of the effect of HPP on lipid oxidation. Total sugars, sucrose, glucose and fructose are not affected by HPP. However sucrose levels dramatically decreased in pressure-treated (600 MPa/25 °C/6 min) raspberry puree, with a corresponding increase in glucose and sucrose levels, most likely due to invertase activity (Butz et al., 2003).

Very few studies are available that have investigated the effect of HPP on dietary fibre. In cabbage, no effect of HPP (up to 500 MPa and 80 °C) on total dietary fibre was evident, however soluble fibre increased, while insoluble fibre decreased at 400 MPa (Wennberg and Nyman, 2004). A similar effect was evident in longan fruit pericarp for water soluble polysaccharides, while other fibre components were not affected by HPP (Yang et al., 2009). HPP has been shown to enhance glucose retardation/binding in tomato puree (Fernandez Garcia et al., 2001; Butz et al., 2002a), suggesting that this technique might be used to develop diabetic foods. Overall, as HPP has a limited effect on covalent bonds, macro- and micro-nutrients tend to not be affected by HPP except at conditions of high pressure and high temperature.

Micronutrients The effect of HPP on vitamins has mainly focussed on ascorbic acid (Vitamin C) content,

although thiamine (B1), riboflavin (B2), pyridoxal (B6) and folate have also been investigated. The majority of these studies have been in fruits and vegetables or model systems. Generally all of these water soluble vitamins are stable to high pressure processing, with folate showing higher sensitivity (Sancho et al., 1999; Indrawati et al., 2002; Oey et al., 2008; Sanchez-Moreno et al., 2009). A summary of recent studies assessing ascorbic acid content is shown in Table 3.

Table 2. The effect of HPP on Macronutrients

Commodity Macronutrient HPP conditions Compared to the experimental control

Reference

Beef Total lipid 0.1, 200 & 800 MPa/60 ºC/20 min

No change Tume et al., 2010

Ham (frozen) Total protein 400 & 600 MPa/~15 °C/10 min

No significant change Serra et al., 2007

Albacore tuna Total lipid 275 & 310 MPa/2, 4, 6 min Significantly ↑ Ramirez-Suarez and Morrissey, 2006

Milk Cheese Cheese – using HP milk

Total lipid Total protein Total lipid Total protein Total lipid Total protein

400 MPa/20 °C/20 min 400 MPa/20 °C/20 min 400 MPa/20 °C/20 min

No change No change No change No change Significantly ↓ Significantly ↑

Sandra et al., 2004

Tomato puree

Glucose binding (retardation)

600MPa/20 °C/60 min 600MPa/25 °C/60min

Significant retention (28 %) ↑ of 14.34-27.13 %

Fernandez Garcia et al., 2001 Butz et al., 2002a

White cabbage

Dietary fibre 0.1, 400 & 500 MPa/20, 50 & 80 °C/10 min

Slight change in TDF. Soluble fibre - ↓ all temp at 400 MPa; Insoluble fibre – concurrent ↑

Wennberg and Nyman, 2004

Longan fruit pericarp Water-soluble polysaccharide Alkali-soluble polysaccharide Cellulose Lignin

0, 200, 300, 400, 500 MPa/25 °C/30 min

↓ with ↑ pressure No significant difference No significant difference No significant difference

Yang et al., 2009

Raspberry puree Sucrose Glucose & fructose

600 MPa/25 °C/6 min 600 & 800 MPa/44 °C/6 min 600 Mpa/25 °C/6 min

>90 % loss during storage at 4 °C for 30 days at 600 MPa 40 % and 60 % ↑ during storage at 4 °C for 30 days at 600 MPa

Butz et al., 2003

Strawberry Raspberry

Sucrose 600 MPa/25 °C/6 min 100 % retained >95 % retained

Butz et al., 2003

Orange juice

Fructose, glucose, sucrose, total sugar Total sugar Sucrose

500 & 800 MPa/20 °C/ min 500 & 800 MPa/20 °C/ min 600 MPa/25 °C/6 min

No difference 100 % retention after 21 days storage (4 °C) 100 % retention

Butz et al., 2002b Butz et al., 2002b Butz et al., 2003

Orange/lemon/carrot mixed juice

Sucrose 600 MPa/25 °C/6 min 100 % retained Butz et al., 2003

Table 3. The effect of HPP on Micronutrients

Commodity Micronutrient HPP conditions* Compared to the experimental control

Reference

Beef Tocopherol Fatty acids

0.1, 200 & 800 MPa/60 ºC/20 min

No change Significant ↑ myristic (14:0); palmitic (16:0); trans-vaccenic acid (18:1trans-11) Significant ↓ linoleic acid (18:2n-6), cis-vaccenic acid (18:1cis-11)

Tume et al., 2010

Iberian ham α-tocopherol 200 & 400 MPa/20 °C/15 min

No effect of HPP ↓ correlation with ↑ TBARS after 39 days storage (5 °C)

Andres et al., 2006

Dry-cured pork loin Total free amino acid 300, 350 & 400 MPa/20 °C/10 min

No change Campus et al., 2008

Orange juice

Ascorbic acid 600 MPa/40 °C/4 min 500 & 800 MPa/20 °C/ min 100 MPa/60 °C/5 min 350 MPa/30 °C/2.5 min 400 MPa/40 °C/1 min

Retained No change. 90% retention after 21 days storage (4 °C) 90 % retention No change 93 % retention. No further loss after 10 days storage (4 °C)

Polydera et al., 2005 Butz et al., 2002b Sanchez-Moreno et al., 2003a

Orange/lemon/carrot mixed juice

600 MPa/25 °C/6 min 600 MPa/25 °C/6 min

95 % retained >95 % retained

Butz et al., 2003 Butz et al., 2003

Tomato juice Carrot juice Tomato puree Carrot puree

Ascorbic acid 250 MPa/35 °C/15 min 400-600 MPa/20 °C/15 min

No change. After 30 days storage (4 & 25 °C) 70 % & 45 % retained respectively 90 % retained

Dede et al., 2007 Patras et al., 2009b

Cowpea Ascorbic acid – 4 days germination 6 days germination

300, 400 & 500 MPa/RT/15 min

90-72% retained 91-59% retained

Doblado et al., 2007

Melon Ascorbic acid 600 MPa/RT/10 min 50 % retained. Cultivar dependent with 15-74 % retention.

Wolbang et al., 2008

Blackberry puree Strawberry puree

Ascorbic acid 400, 500 & 600 Mpa/10-30 °C/15min

No significant change (>90 % retained)

Patras et al., 2009a



Ascorbic acid has been shown to be stable to high pressure processing, unless it is subjected to high pressure and high temperature (>65 °C) conditions, where oxidation reactions are enhanced (Oey et al., 2008). In addition to its role as a vitamin, ascorbic acid also plays an important role as an antioxidant. Ascorbic acid has been shown to protect folate against the effects of pressure and heat (Oey et al., 2008). Further information about the antioxidant role of ascorbic acid can be found in the section on HPP and antioxidant activity.

Most studies of the effect of HPP on fat soluble vitamins have been limited to carotene, Vitamin A (retinol) and Vitamin E (tocopherol). Again, HPP has minimal effect on these vitamins (Table 2) (Indrawati et al., 2002; Oey et al., 2008). The effect on carotenoids is expanded upon in the section on antioxidant activity.

There is still limited information available about the effect of HPP on niacin, cobalamin, and Vitamins A, D, E and K.

Information about the effect of HPP on minerals is scarce, with most discussion of minerals focussing on possible relationships between iron and lipid oxidation. This is an area that requires further investigation.

As with the other micronutrients mentioned, few studies have been reported that investigate amino acid or fatty acid content except in meat products. HPP (up to 400 MPa) of dry-cured pork loin has no effect on the total free amino acid content (Campus et al., 2008). Some variation in individual fatty acids occurs when beef is subjected to HPP (up to 800 MPa) (Tume et al., 2010). Further investigation into the effect of HPP on individual fatty acids is necessary, especially in relation to essential fatty acid content.

Proteins and Enzymes Proteins are usually denatured by high pressure however the protein type, processing

conditions and pressures applied are all important considerations. Generally, at low protein concentrations and low pressures (<300 MPa), reversible pressure-induced denaturation occurs, while higher pressures (>300 MPa) induce irreversible and extensive effects on proteins (Guerrero-Beltran et al., 2005; Rastogi et al., 2007). HPP is most commonly used to inactivate deleterious enzymes, thereby ensuring the high quality characteristics of the food are maintained (Rastogi et al., 2007), but it can also be used to stabilise and activate other enzymes (Eisenmenger and Reyes-de-Corcuera, 2009).

These changes tend to have little effect on the nutritional content of the food, but are important with respect to food quality especially in relation to colour (polyphenoloxidase) and texture of food (pectic enzymes), plus some influence lipid oxidation (lipase and lipoxygenase). Thus most investigations have focused on monitoring the effect of high pressure on these quality related enzymes, and there are a number of comprehensive review articles available summarising this work (Guerrero-Beltran et al., 2005; Rastogi et al., 2007; Eisenmenger and Reyes-de-Corcuera, 2009). Table 4 shows a summary of the effect of HPP on proteins and enzymes in various commodity groups. Future work may focus on expanding the knowledge of HPP on antioxidant enzymes as another means of maintaining food quality and nutrition.

Table 4. The effect of HPP on Proteins

Commodity Protein type HPP conditions* Compared to the experimental control

Reference

Cows Milk Casein miscelle

250 MPa >400 MPa

Size temp dependant Size altered

San Martin-Gonzalez et al., 2006

Whey proteins β-lactoglobulin α-lactalbumin BSA

500 & 800 MPa 500 & 800 MPa ≤400 MPa/RT/60 min

70 % & 90 % denatured 10 % & 50 % denatured Not denatured


Goats milk Casein miscelle Lipase

300-350 MPa / 45 °C 400-500 MPa 300-400 MPa/0-180 min

↑ Size Smaller size ↑ activity


Soy milk Lipoxygenase >800 MPa 600 MPa/60 °C

Inactivated Inactivated

Van der Ven et al., 2005

Cold-smoked salmon Proteolytic enzymes (cathepsin and calpains)

≤300 MPa/9°C/20 min Reduced activity Lakshmanan et al., 2005

Lupin Meta (bovine)

Protein digestibility (in-vitro)

200 & 500 MPa/10 °C/10 min

Most hyrolysed at 500 MPa De Lamballerie-Anton et al., 2002

Ham (frozen) Antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidise) Proteolytic enzymes (cathepsin B, cathepsin B+L)

400 & 600 MPa/~15 °C/10 min

Significant ↓, but still active No significant change

Serra et al., 2007

Dry-cured pork loin Cathepsins amino-peptidases dipeptidyl peptidases

300, 350 & 400 MPa/20 °C/10 min

20 % ↓ for 400 MPa; Significant ↓ after 45 days storage Significant ↓ Significant ↓

Campus et al., 2008


Table 5. The effect of HPP on Lipid Oxidation

Commodity Lipid oxidation test HPP conditions* Compared to the experimental control

Reference

Chicken (minced thighs) TBARS (to 9 days) TBARS (7 days)

500 MPa/50 ºC/30 min (air storage) 500 MPa/-10, 5, 20, & 50 ºC/30 & 60 min (vacuum stored) 0.1, 200, 400, 600 & 800 MPa/20-70 °C/ 20 min

Raw – significant ↑ at 6 & 9 days Overcooked – significant ↑ from day 1 Raw – no significant difference after 9 days ↑ at >400MPa regardless of temperature

Beltran et al., 2004 Ma et al., 2007

Beef TBARS (up to 6 days) Peroxide Value TBARS (7 days)

0.1, 200 & 800 MPa /60 ºC/20 min 0.1, 200, 400, 600 & 800 MPa/20-70 °C/ 20 min

↑ in 6 day samples Low at 200 MPa ↓ at 6 days At 20 & 40 °C - 5 fold ↑ at ≥400MPa. Little additional effect at > temperatures

Tume et al., 2010 Ma et al., 2007

Iberian ham TBARS (90 days storage) TBARS (39 days)

200-300 MPa/20-30 min 200 & 400 MPa/20 °C/15 min + MAP

No differences Significant ↑ at 39 days (5 °C) for 400 MPa No O2 = low TBARS

Andres et al., 2006

Dry-cured pork loin

TBARS (up to 45 days storage)

300, 350 & 400 MPa/20 °C/10 min

No difference Campus et al., 2008

Ovine Milk

Free Fatty Acid value 500 MPa /4, 25, 40 °C No change or ↓ Norton and Sun, 2008

Rainbow trout (red muscle)

TBARS (up to 6 days)

150, 300, 450, 600 MPa/ 5 min

No change at 150 MPa ↑ with ↑ pressure 300 MPa ideal

Yagiz et al., 2007

Mahi Mahi (red muscle)

TBARS (up to 6 days)

150, 300, 450, 600 MPa/RT/15 min

↑ with ↑ pressure, with max at 300MPa then ↓; 450 MPa ideal

Yagiz et al., 2007

Sardine (cold smoked) TBARS & Peroxide Value

300 MPa/20 ºC/20 min Little change over 2 wk storage (air)

Gomez-Estaca et al., 2007

Albacore tuna (minced muscle)

TBARS (22 days/4 °C & 93 days/-20 °C)

275 & 310 MPa/2, 4, 6 min Lower immediately after HPP. Slight increase with storage

Ramirez-Suraz and Morrissey, 2006



Lipid Oxidation The effect of HPP on lipid oxidation is important from a nutritional perspective due to the

loss of polyunsaturated lipids and the production of undesirable oxidation products. Most studies have focussed on meat, some on seafood, and a few on dairy products. The focus on muscle foods is primarily due to reports that lipid oxidation is enhanced at processing pressures >400 MPa and is more prevalent in red meat due to the higher myoglobin levels (Ma et al., 2007). It has also been suggested that free metal ions are released by the pressure treatment, again promoting lipid oxidation. Some recent studies investigating lipid oxidation in HPP foods are summarised in Table 5 and they reiterate the importance of keeping the processing pressure at <400 MPa to reduce lipid oxidation. However they also highlight that by vacuum storing the pressure processed muscle food, lipid oxidation is minimised (Beltran et al., 2004; Andres et al., 2006). In the case of fish, lower processing pressures seem to result in lower levels of lipid oxidation. The actual effect on individual fatty acids, particularly polyunsaturated fatty acids requires further investigation.

EFFECTS ON BIOACTIVE COMPONENTS IN FOODS Food commodities or ingredients that provide health-related benefits beyond their basic

nutrient supply have gained wide spread acceptance by consumers. There is a vast range of functional foods in terms of their chemical nature, target biomarker and target health benefit. The beneficial effects of foods are associated with the bioactive components present in them including plant pigments (anthocyanin, carotenoids), phenolic compounds, vitamins, fatty acids and peptides. Current trends in the functional food area are towards retaining the maximum level of bioactive components while maintaining fresh-like qualities of foods. With this in mind studies have focused on the effects of HPP on carotenoids, total phenolic compounds, anthocyanins and flavonones mainly in fruits and vegetables and in a few other commodities, as detailed in Table 6. Among the wide range of bioactive components in fruits and vegetables, HPP treatment has shown remarkable benefits in retaining or increasing the levels of total carotenoids in foods (Table 6). A significant increase in all types of carotenoids has been observed in tomato puree and orange juice (Sanchez-Moreno et al., 2005 & 2006), papaya slices (De Ancos et al., 2007) and melon (Woolbang et al., 2007) around pressures of 400-600 MPa. Broccoli and carrots have retained a maximum of 95-100% of α- and β-carotene at similar pressures, while lycopene (tomato puree) and lutein (in broccoli and green beans) also showed very high stability (100% retention) after HPP.

The effect of HPP on anthocyanin and total phenolic compounds in berry fruits and a few tropical fruits has also been studied. Anthocyanin in strawberry, raspberry and blackberry juices after HPP showed 100% retention (400-600 MPa at room temperature) and increased pressures led to an increase in stability. However, the retention percentage varied in different fruit matrices being 54% in muscadine grape juice (550 MPa) (Del Pozo-Insfran et al., 2007) and 50-42% in blackcurrant (800 MPa) (Kouniaki et al., 2004). Anthocyanin retention during HPP seems to relate to other phenolic compounds, vitamin C content and PPO activity in fruits. Elevated temperatures during HPP and storage conditions seem to reduce the anthocyanin content. Anthocyanin present in HPP strawberry, blackcurrant and raspberry was

Trends in High Pressure Processing of Foods 135

most stable when a pressure of 800 MPa for 15 min was applied (Tiwari et al, 2009) however, anthocyanin degradation in HPP processed juice was observed when HPP combined with heat. This decrease could be related to condensation reactions with other flavanols or organic acids in juices, and better stability at higher pressures may be attributed to complete inactivation of the PPO enzyme (Tiwari et al., 2009).

Other phenolic compounds such as phenolic acids and flavonoids seem highly stable or in fact increased during HPP. A significant increase in total phenolics has been reported in strawberry (9%), blackberry (5%), onion (12%) and longman fruit, while litchi had no significant changes (Table 6). High pressure enhances mass transfer rates which increase cell permeability leading to an increased extraction of cellular components and increasing levels of pigments in the juice. Small molecules such as pigments and volatile flavour compounds connected with the sensory quality of foods are unaffected by HPP (Cheftel, 1995).

HPP treated (400 MPa/40 °C/1 min) orange juice (which is a very rich source of flavanone glycosides which are degraded to aglycones by human intestinal flora after ingestion) showed an increase in levels of naringenin (20%) and hesperetin (39%), but insignificant changes at 4 ºC for 10 days storage. It is suggested that at around 400 MPa, some structural changes in the cell walls of the orange juice sacs may have led to a release of phenols from protein, and consequently an increase in the extraction of flavanones. (Sanchez-Moreno et al., 2005).

A study on the effect of HPP on isoflavones (complex glucosides and bioavailable forms of aglycone) in soybean seeds and soymilk reported insignificant changes to total isoflavone content and better retention of isoflavones compared with conventional thermal processing (Jung et al., 2008). However, the isoflavone profile was significantly modified due to the adiabatic heating occurring during pressurization combined with mild temperature treatment.

Apart from fruits and vegetables, limited studies have also focused on bioactive components in dairy (immunoglobulin, lactoglobulin and lactoferrin) and fish (omega-3 fatty acids). Immunoglobulin A in human milk and cow’s milk showed better stability at 400 MPa with retentions of 75-86% (Viazis et al., 2007). However, significant changes in immunoglobulin protein in goat’s milk has been reported at 500 MPa and in bovine colostrum at 400 MPa and above, due to the denaturation of proteins. Pressures of 200 MPa applied for 2 hours may be used to retain at least 85% of the immunoglobulin activity, although the microbial load of colostrum is reduced by less than 2-log cycles which is insufficient to maintain the shelf-life of the product (Palmano et al, 2010). Lactoferrin, a bioactive milk peptide, seems to be highly pressure stable at pH 4.5 around 400 MPa. Yoghurt with added lactoferrin showed 80% retention of Lactoferrin after HPP (Carroll et al., 2010).

EFFECTS ON ANTIOXIDANT ACTIVITY (AA) Numerous plant constituents, such as phenolic compounds, vitamin C and carotenoids in

fruits and vegetables, have been recognised as natural antioxidants that possess beneficial effects against free radical related oxidative damage associated with a number of diseases (Prasad et al., 2009). The pressure stability of antioxidants present in fruits and vegetables compared to other conventional processes is of great interest.

Table 6. The effect of HPP on Bioactive Components

Commodity

Bioactive compound

HPP conditions

Compared to the experimental control

Reference

Tomato Puree Tomato puree

Total Carotenoid (mgl-1) α carotene β carotene Lycopene Lutein Vitamin A(RAE/litre) Lycopene

400 MPa/25 °C/15 min 600 MPa/25 °C/ 60 min 400 MPa/25 °C/ 15 min

Significant ↑ in all types 82% 36% 48 % 71% 39% 100% retention 49% ↑

Sanchez-Moreno et al., 2006 Butz et al., 2002 Sanchez-Moreno et al., 2006

Carrots (Whole) Carrot Juice

α carotene and β carotene Carotenoids

400-600 MPa/25 °C/2 min 300 MPa/ 50 °C

100% retention Highest stability

McInerney et al., 2007 Kim et al., 2001

Broccoli (Whole)

α carotene and β carotene Lutein

400-600 MPa/25 °C/2 min 95- 83% retention 100-90% retention

McInerney et al., 2007

Green beans (Whole)

Lutein 400-600 MPa/25 °C/2 min 100% retention McInerney et al., 2007

Onion

Total phenol content Flavonol content Total quercetin

100 MPa 50 °C/5 min 400 MPa/5 °C/5 min 100-400 Mpa/5 °C/5 min

12% ↑ 26% ↑

Roldan-Marin et al., 2009

Orange juice

Zeaxanthin Lutein α carotene β carotene β carotene Flavonones

400 MPa/40°C/1 min 600/MPa /20°C/1 min storage 4 & 10°C 100-450 MPa /30-40°C /1-2.5 min

% ↑ 44 75 30 33 No significant difference after HPP and storage ↑ at 350-450 MPa

Sanchez-Moreno et al., 2005 Bull et al., 2004 Sanchez-Moreno et al., 2005

(mg/100g DW) Total phenols Anthocyanin Anthocyanin Anthocyanin

400-600 MPa/20 °C/15 min 800 MPa/18-22 °C/15 min 200- 800 MPa/20 °C/15 min

↑ 9.8% in strawberry and 5% in blackberry at 600 MPa 100% retention in both No significant change after HPP Greater stability at 800 MPa

Patras et al., 2009b Garcia-Palazon et al., 2004 Sulthangjai, et al., 2005, Zabetakis, et al.,

Table 6. (Continued)

Commodity

Bioactive compound

HPP conditions

Compared to the experimental control

Reference

Blackcurrent

Anthocyanin

200-800 MPa/20 °C/15 min

Little change soon after HPP at 600 MPa; 70% retention after five days in storage (4 °C). 50-58% ↓ at 800 MPa

2000 Kouniaki, et al., 2004

Muscadine grape Juice Anthocyanin 400 & 550 MPa/15 min Better retention at higher pressures. 30% at 400 MPa and 54% at 550 MPa

Del Pozo-Insfran et al., 2007

Melon

β carotene

600 MPa/20 °C/10 min

Significant ↑ Woolbang et al., 2008

Papaya slices

Caroteinoids 400 MPa/25 °C/1 min

56 % ↑ De Ancos et al., 2007

Longman fruit Total phenolics (TP) 200-500 MPa/30°C/2.5- 30 min

Significant ↑ as pressure ↑. Higher extraction of TP in HP extracted juice than solvent extracts. Time had no significant effect

Prasad et al., 2009a

Litchi fruit

Total Phenolics 200-500 MPa/30°C/2.5- 30 min

No significant difference. Prasad et al., 2009c

Soymilk

Isoflavone 300-700 MPa No significant change in total isoflavone at 750 MPa/25 °C Glucoside content ↑, total aglycon content ↓

Jung, et al., 2008

Human milk

Immunoglobulin A 400 MPa/30 °C/30-120 min

Retention 86-75% from 30-120 min.

Viazis, et al., 2007

Yogurt

Lactoferrin (added)

HPP treatment (500 MPa) for milk before yogurt preparation

80% retention Carroll, et al., 2010

Salmon (Atlantic)

Omega 3 n-3 PUFA and n-6PUFA

150-300MPa/15min No significant difference after HPP. Greater retention than cooking at 72 °C

Yagiz, et al., 2009

Table 7. The effect of HPP on Antioxidant Activity Commodity Method used HPP conditions Compared to the

experimental control Reference

Carrots Whole Puree Puree Juice Juice

FRAP (Fe+2/Kg) TEAC index Anti Radical Power (g/l-1) TEAC index DPPH Radical scavenging activity

400-600 MPa/25 °C/2 min 800 MPa/20 °C/5 min 400-600 MPa/250 °C/15 min 100-800 MPa/30-65 °C/max 90 min 150-250 MPa/5-35 °C/5-15 min

Modest ↓ at 400 but no difference at 600 MPa Slight but significant difference 33% ↑ at 600 MPa ↑ by HPP but ↓ above 40 °C at higher pressures No significant difference up to 250 MPa/35°C/15 min. Significant but slight change after that and during storage (80% retention)

McInerney et al., 2007 Butz et al, 2002 Patras et al, 2009a Indrawati et al, 2004 Dede et al, 2007

Tomato puree Puree Puree

DPPH Radical scavenging activity TEAC Index Anti Radical Power

400 MPa/25 °C/15 min 500-800 MPa/20 °C/5 min 400-600 MPa/25 °C/15 min

Significant ↓ Insignificant ↓ at 800 MPa Significant ↑

Sanchez –Moreno et al., 2006 Butz et al., 2002 Patras et al., 2009a

Puree ABTS+ (TEAC index) 500-800 MPa/20 °C/5min Minimal initial loss Fernandez Garcia et

Juice

DPPH Radical scavenging activity

150-250 MPa/5-35 °C/5-15 min

(70% retained) No significant difference up to 250 MPa/25°C/15 min.

al., 2001 Dede et al., 2007

Broccoli

FRAP (Fe+2/Kg) 400-600 MPa/25 °C/2 min No significant effect McInerney et al., 2007

Green Beans

FRAP (Fe+2/Kg) 400-600 MPa/25 °C/2 min Water soluble antioxidants ↑ at both levels

McInerney et al., 2007

Onion


100 MPa/50 °C/5 min 400 MPa/5 °C/5 min

↑ trend with ↑ pressures. No significant effect at 400 MPa/5 °C

Roldan-Marin et al., 2009

Cowpea (Legume) Raw Germinated

TEAC Index

300- 500 MPa/room temperature/15 min

Slight ↓ 90-85% retention 80% retention

Doblado et al., 2007

Orange Juice

TEAC Index TEAC Index ABTS+ (TEAC index) ABTS

800 MPa/20 °C/5min 100-600 MPa/65°C/90min 500-800 MPa/20 °C/5min 600 MPa/40 °C/4 min

No great difference after HPP; 10% loss during three weeks storage ↓ with ↑ pressure 85% retention after 21 days storage at 4 °C Better retention of

Butz et al., 2002 Indrawati et al., 2004 Fernandez Garcia et al., 2001 Polydera et al., 2005

Table 7. (Continued) Commodity Method used HPP conditions Compared to the

experimental control Reference


100-450 MPa/30-40 °C/1-2.5 min

antioxidant activity than thermal treatment No change after HPP. 95% retention at 4 °C & 10 days storage

Sanchez-Moreno, et al., 2003a

Berries Strawberry Blackberry

Free radical scavenging activity as Anti radical power ARP(g/l-1)

400- 600 MPa/20 °C/15 min

Significant ↓ 67% ↑ at 600 MPa

Patras et al., 2009b

Apple and Peach

TEAC Index µmol/ml

60 MPa No significant change Butz et al., 2003

Longman fruit DPPH Radical scavenging activity

200-500 MPa/30 °C/2.5- 30 min

↑antioxidant activity in HP extracted juice compared with solvent extracts

Prasad et al., 2009a Prasad et al., 2009 b

Litchi fruit


200-500 MPa/30 °C/2.5- 30 min

No significant change Prasad et al., 2009c


Recent research has mainly focused on a few commodities such as orange juice, carrot puree and tomato puree, while limited studies have focused on broccoli, green beans, onion, cowpea, berry fruits, apples, litchi and longman fruit using a range of analytical methods to test the antioxidant capacity as detailed in Table 7.

In general, the effect of HPP on the antioxidant activity is not the same among the food products and depends on the vitamin stability and the extraction yield of existing bioactive compounds, quality related enzymes, such as PPO levels, and pH conditions of the matrices. Antioxidant activity of several fruits and vegetables showed insignificant changes after the application of pressures around 600-800 MPa (Butz et al., 2003).

Orange juice showed a good retention of ascorbic acid after HPP around 500-800 MPa at room temperature and for shorter time periods (5 min) however, a slight reduction (10-15%) has been reported during three weeks storage at 4 °C. As the exposure time increased (90 min) the reduction of ascorbic acid accelerated at higher pressures (Indrawati et al., 2004).

Among compounds exhibiting antioxidant activity in orange juice, vitamin C is the most important accounting for 68-90% of total antioxidant capacity (Polydera et al., 2005), therefore the retention of vitamin C during HPP is more important than the other compounds. Carrots and tomato showed a small reduction in ascorbic acid in most of the studies except in one study where a significant increase of 33% at 600 MPa was found, which was reflected by the increased levels of carotenoids and better retention of vitamin C after HPP (Patras et al., 2009a). High pressure processed fruit purees had significantly higher antioxidant capacities when compared to thermally treated samples (Patras et al., 2009b).

OTHER HEALTH RELATED EFFECTS In recent years attention has also been paid to potential anticarcinogenic components

from fruits and vegetables. Studies have also reported the retention of higher levels of anticariogenic (Prasad et al., 2009c), allergenic (Indrawati et al., 2002) and anti-inflammatory properties of HPP foods. To date, there are no published reports about toxicity in HPP foods. Further investigations into both the effects of HPP on allergenicity and toxicity are necessary.

Reports show that high hydrostatic pressure enhances whey protein digestibility to generate whey peptides that improve anti-inflammatory activity. The altered profile of low molecular weight peptides, isolated from hydrolysates of pressurized (400 & 550 MPa, 3-4 min) whey proteins, has indicated a strong upward trend in the total and reduced GSH content in cultured mutant CFTR cells in comparison to peptides released from native whey hydrolysates (Vilela et al., 2006). In another study, HPP orange juice (400 MPa/40°C/1 min) consumption improved the plasma vitamin C, antioxidative status and inflammatory markers in healthy humans (Sanchez-Moreno et al., 2003b)

In-vitro experimental systems also show that flavonoids possess anti-inflammatory, anti-allergic, antiviral, hypocholesterolemic, and anticarcinogenic properties. Even though there is not enough research with HPP in relation to each and every healthful property of foods, the retention or increased levels of flavones in orange juice and soybeans (healthful properties of isoflavones such as lowering cholesterol levels, preventing both prostate and breast cancers, attenuating bone loss in postmenopausal women and alleviating menopausal symptoms), carotene and lycopene levels in carrots and tomatoes, predict the benefits of HPP in


maintaining or enhancing the healthful attributes of a wider range of commodities. As a consequence, HPP can be regarded as an alternative or complementary technology to thermal processing.

CONCLUSION From a nutritional perspective, high pressure processing is an excellent food processing

technology which has the potential to retain compounds with health properties. Macronutrients and most micronutrients do not appear to be affected by HPP. Careful selection of pressures around 400-600 MPa and shorter exposure times (5-15 min) are essential to retain maximum levels of bioactives which exert antioxidant, anti-carcinogenic and anti-inflammatory properties. These conditions are also recommended to minimise the incidence of lipid oxidation in food after HPP. Lower pressures of around 100 MPa have little effect however, pressures around 400 MPa enhance the cell permeability and structural changes leading to improved extraction of nutrients and bioactive components in food matrices. HP treatment at extreme pressure and temperature combinations could result in degradation of vitamin and bioactive components

LIMITATIONS AND RECOMMENDATION This chapter has revealed a number of limitations in current knowledge of the effect of

HPP on particular compounds in foods. Generally, investigations into the effect of HPP on dietary fibre have been fairly limited, suggesting the greatest effect is on water soluble fibre components. Further investigations using a broader variety of foods would be useful, especially as dietary fibre is considered to play an important role in human health. In addition, the pressure stability of the B-group vitamins, particularly niacin and cobalamin, the fat soluble vitamins E and D, and minerals is currently almost unknown. It is known that lipid oxidation can be induced following HPP, yet few studies have investigated the effect on individual fatty acids, particularly essential fatty acids in foods. An expanded knowledge of the effect of HPP on antioxidant enzymes would help in maintaining food quality. Information about how HPP affects toxins is rare, as is information about the possible allergenicity of HPP food. The fate of antioxidants in other HPP fruits and vegetable products, dairy, fish and meat products, is another area requiring investigation.

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