A comparison between high hydrostatic pressure extraction and heat extraction of ginsenosides from ginseng (Panax ginseng CA Meyer)

BACKGROUND: To determine biomaterial components, the components must first be transferred into solution; thus extraction is the first step in biomaterial analysis. High hydrostatic pressure technology was used for ginsenoside extraction from ginseng roots. In the extraction of fresh and red ginseng, high hydrostatic pressure extraction (HHPE) was found to be more effective than heat extraction (HE). RESULTS: In fresh ginseng extraction under HHPE, total ginsenosides (1602.2 µg mL^?1) and ginsenoside metabolite (132.6 µg mL^?1) levels were slightly higher than those under HE (1259.0 and 78.7 µg mL^?1), respectively. In red ginseng, similar results indicated total ginsenoside and ginsenoside metabolite amounts according to the extraction methods. Most volatile compounds by HHPE were higher than by HE treatment. HHPE of red ginseng was conducted under four pressures: 0.1 MPa (1 atm), 30, 50, and 80 MPa. Total sugar, uronic acid, and polyphenol amounts increased until 30 MPa of pressure and then showed decreasing tendencies. Total ginsenoside and ginsenoside metabolite contents linearly increased with increasing pressure, and a maximum was reached at 80 MPa for the metabolites. CONCLUSION: HHPE used for red ginseng processing contributes to enhanced extraction efficiencies of functional materials such as ginsenosides through cell structure modification. Copyright © 2011 Society of Chemical Industry     

Effects of high hydrostatic pressure on enzymes, phenolic compounds, anthocyanins, polymeric color and color of strawberry pulps

BACKGROUND: Changes in activity of polyphenol oxidase (PPO), peroxidase (POD) and β‐glucosidase, individual phenolic compounds other than anthocyanins, total phenols, monomeric anthocyanins, polymeric color and instrumental color of strawberry pulps were assessed after high hydrostatic pressure (HHP) (400–600 MPa 5–25 min^?1) at room temperature. RESULTS: β‐Glucosidase was activated by 4.7–16.6% at 400 MPa 5–25 min^?1 and inactivated by 8.0–41.4% at 500 or 600 MPa. PPO and POD were inactivated at all pressures, the largest reduction in activity being 41.4%, 51.5% and 74.6%, respectively. The individual phenolic compounds and total phenols decreased at 400 MPa, but total phenols increased at 500 or 600 MPa. However, the monomeric anthocyanins, polymeric color and redness (a*) exhibited no change. HHP induced a decrease in lightness (L*) and an increase in yellowness (b*) at 400 MPa, but no significant alteration in L* value and b* value at 500 or 600 MPa was observed; this was attributed to higher residual activity of PPO, POD and β‐glucosidase at 400 MPa. Total color difference (ΔE) was ≥ 5 at 400 MPa and ?3 at 500 or 600 MPa. CONCLUSION: HHP effectively retained anthocyanins, phenolic compounds and color of strawberry pulps, and partly inactivated enzymes. Copyright © 2011 Society of Chemical Industry     

Characterization of red ginseng residue protein films incorporated with hibiscus extract

An edible film was prepared from red ginseng residue protein (RGRP) and incorporated with hibiscus extract (HE). RGRP was extracted from red ginseng residue, which is an inexpensive by-product of the red ginseng processing industry. Different concentrations of HE were added to an RGRP film-forming solution as a natural antioxidant. The prepared RGRP films without HE had a tensile strength of 16.9 MPa and an elongation at break of 25.1%. The antioxidant activity of the RGRP film increased with increasing concentration of HE. In addition, the RGRP film with 1% HE exhibited the lowest value of water vapor permeability (1.88×10^?9 g·m/m²·s·Pa), which indicates that the film has high water barrier property. The results present the production of edible films from discarded red ginseng residue, and the antioxidant activity of RGRP films as a packaging material can prevent lipid oxidation and quality loss of food products.     

Application of High Hydrostatic Pressure and Enzymatic Hydrolysis for the Extraction of Ginsenosides from Fresh Ginseng Root (Panax ginseng C.A. Myer)

The effect of high hydrostatic pressure (HHP) processing combined with enzymatic hydrolysis for the extraction of ginsenosides from fresh ginseng roots (Panax ginseng C.A. Myer) was evaluated. Fresh ginseng roots were subjected to cellulase and β-amylase hydrolysis for 12 h with HHP (100 MPa) at 50 °C. The use of high hydrostatic pressure and enzymatic hydrolysis (HHP-EH) increased the yield of total saponins, panaxadiols, and metabolites from fresh ginseng roots. The yield of total saponin was 40.2 mg/ml from the HHP-EH with cellulase treatment, significantly higher (P?<?0.05) than the yield from the HHP-EH with β-amylase treatment (36.1 mg/ml). The amounts of Rg1?+?Rb1, panaxadiols, panaxatriols, and metabolites obtained from the HHP-EH with cellulase treatment were 5.56, 12.05, 9.78, and 1.88 mg/ml, respectively. These values were significantly higher compared to those obtained from heat extraction and HHP-EH with β-amylase treatment. In addition to ginsenosides, studies on the extraction of other active ingredients by HHP-EH processing are warranted.     

Drying characteristics and quality of taegeuk ginseng (Panax ginseng C.A. Meyer) using far‐infrared rays

The purpose of this study is to investigate the drying characteristics of taegeuk ginseng using a far‐infrared dryer. The dryer used for this experiment can control the drying parameters such as far‐infrared heater temperature and air velocity. The far‐infrared drying tests of taegeuk ginseng were performed using large, medium and small ginseng at an air velocity of 0.6 m s^?1, and drying chamber temperatures of 45, 55 and 65 °C. The results were compared with those of the hot‐air drying method. The drying characteristics were analysed based on factors such as drying rate, colour changes, internal tissue structure of ginseng, saponin contents and antioxidant activities, and energy consumption. In general, the hot‐air drying time was 3–3.5 days longer than that of far‐infrared drying under the same conditions. The colour changes of far‐infrared drying were also smaller than that of hot‐air drying. Far‐infrared dryer required less energy consumption about 9.67–14.8% less than that required by hot‐air drying. At the same time, the saponin and polyphenol contents of taegeuk ginseng subjected to far‐infrared drying were 1.01 mg g^?1, 7.81 mg/100 g higher than that subjected to hot‐air drying, respectively. The results showed that the far‐infrared drying increases the capacity and quality of taegeuk ginseng.     

Inactivation of Vibrio parahaemolyticus in Hard Clams (Mercanaria mercanaria) by High Hydrostatic Pressure (HHP) and the Effect of HHP on the Physical Characteristics of Hard Clam Meat

Shellfish may internalize dangerous pathogens during filter feeding. Traditional methods of depuration have been found ineffective against certain pathogens. The objective was to explore high hydrostatic pressure (HHP) as an alternative to the traditional depuration process. The effect of HHP on the survival of Vibrio parahaemolyticus in live clams (Mercanaria mercanaria) and the impact of HHP on physical characteristics of clam meat were investigated. Clams were inoculated with up to 7 log CFU/g of a cocktail of V. parahaemolyticus strains via filter feeding. Clams were processed at pressures ranging from 250 to 552 MPa for hold times ranging between 2 and 6 min. Processing conditions of 450 MPa for 4 min and 350 MPa for 6 min reduced the initial concentration of V. parahaemolyticus to a nondetectable level (<10¹ CFU/g), achieving >5 log reductions. The volume of clam meat (processed in shell) increased with negligible change in mass after exposure to pressure at 552 MPa for 3 min, while the drip loss was reduced. Clams processed at 552 MPa were softer compared to those processed at 276 MPa. However, all HHP processed clams were found to be harder compared to unprocessed. The lightness (L*) of the meat increased although the redness (a*) decreased with increasing pressure. Although high pressure‐processed clams may pose a significantly lower risk from V. parahaemolyticus, the effect of the accompanied physical changes on the consumer's decision to purchase HHP clams remains to be determined. Practical Application : Shellfish may contain dangerous foodborne pathogens. Traditional methods of removing those pathogen have been found ineffective against certain pathogens. The objective of this research was to determine the effect of high hydrostatic pressure on V. parahaemolyticus in clams. Processing conditions of 450 MPa for 4 min and 350 MPa for 6 min reduced the initial concentration of V. parahaemolyticus to a nondetectable level, achieving >5 log reductions.     

Effects of Combined High Hydrostatic Pressure and Dense Phase Carbon Dioxide on the Activity,Structure and Size of Polyphenoloxidase

High hydrostatic pressure (HHP) may activate undesirable enzymes such as polyphenoloxidase (PPO). Carbon dioxide (CO₂) addition to HHP could increase enzyme inactivation. We investigated the inactivation of combined HHP and dense phase carbon dioxide process on activity, secondary conformation and size of pure PPO from mushroom. Solutions (2.35μM, in phosphate buffer pH 6.8) were treated with HHP alone (HHP), or 3.6% w/w of CO₂ was injected into the package (HHP+CO₂). Treatment conditions were 600 MPa, 20 °C, for 1, 3, 5, 7, and 9 min. HHP+CO₂ treatment significantly decreased residual enzyme activity (REA) to 30% to 12% after 1 to 9 min, respectively, whereas only HHP had no significant effect. Both HHP and HHP+CO₂ treatments caused changes in secondary conformations, however HHP+CO₂ changes were more extensive. Alpha‐helix fractions were reduced by 32% and 41%, while β sheet, turn and unordered increased by 63% and 213%, 100% and 71%, and 118% and 82% for HHP and HHP+CO₂, respectively after 9 min. The protein size in HHP+CO₂ samples was 5‐ to 6‐fold larger than that of Control and HHP treatment, and this increase was inversely correlated with REA. The best inactivation kinetics of HHP+CO₂ model was the 2‐fractional model with 2 simultaneous 1st‐order steps, contributing 70% and 30% to original enzyme activity, with k_labile = 12.15 min^?1 and k_stable = 0.07 min^?1, respectively. No recovery in activity, secondary conformation and size in all samples were observed after 1‐mo storage. Addition of CO₂ in HHP treatment can improve enzyme inactivation, and therefore product shelf‐life and quality.     

Effect of High Hydrostatic Pressure (HHP) Treatment on Physicochemical Properties of Horse Mackerel (Trachurus trachurus)

The basic objective of this study was to determine the effect of high hydrostatic pressure (HHP; 220, 250 and 330 MPa), holding time (5 and 10 min) and temperature (7, 15 and 25 °C) on some quality parameters of horse mackerel such as colour changes, thiobarbituric acid (TBA-i) and trimethylamine nitrogen (TMA-N), free amino acid content. HHP increased L ^* values of horse mackerel. The a ^* and b ^* of treated horse mackerel did not change significantly after HHP applications. After, HHP, TBA-i and TMA values of all HHP-treated horse mackerel samples remained unchanged than those of untreated samples. The results obtained from this study showed that the quality of high pressure treated horse mackerel is best preserved at 250 MPa, 7–15 °C for 5 min, 220 MPa, 15–25 °C for 5 min, 250 MPa, 15 °C for 10 min and 330 MPa, 25 °C for 10 min.     

Cold pasteurisation of red wines with high hydrostatic pressure to control Dekkera/Brettanomyces: effect on both aromatic and chromatic quality of wine

This paper reports the effect of high hydrostatic pressure (HHP) on wines contaminated with Dekkera/Brettanomyces populations of 10⁴ and 10⁶?cfu?mL^?1 growing at either pH 3.2 or 3.6 (both normal pHs for red wine) and at room temperature (25?°C). HHP (100?MPa for 24?h) was highly effective at controlling the growth of all combinations of starting yeast population and pH, yet caused scant modification of thermosensitive wine molecules such as pigments and volatile compounds (molecules that greatly influence wine quality). The present results support the potential use of HHP as a means of cold-pasteurising wines to control Dekkera/Brettanomyces.     

Effects of high hydrostatic pressure on color of spinach purée and related properties

BACKGROUND: Changes in instrumental color parameters, chlorophyll a and b, activity of chlorophyllase, Mg‐dechelatase, peroxidase and polyphenol oxidase, total phenolic compounds and pH of spinach purée were assessed after high hydrostatic pressure (HHP) (200, 400 and 600 MPa for 5, 15 and 25 min) treatments at room temperature. RESULTS: HHP treatments induced a better retention of visual green color (?a* and L* values) and chlorophyll contents of spinach purée. As for chlorophyll degradation‐related enzymes, the results indicated that chlorophyllase activity decreased at all pressures; however, Mg‐dechelatase activity was dramatically activated after HHP treatment at 400 and 600 MPa. Peroxidase exhibited higher resistance to HHP; however, polyphenol oxidase, which is responsible for enzymatic browning, was suppressed progressively with increase in pressure level from 200 to 600 MPa. In addition, the pH value of HHP‐treated spinach purée was increased to be close to neutral pH, which could effectively inhibit chlorophyll degradation. No significant differences (P > 0.05) were found after extending the treatment times at the same level of pressure. CONCLUSION: HHP treatments effectively prevent chlorophyll degradation and enzymatic browning in spinach purée and retain a better original fresh green color of spinach compared with conventional thermal treatment. Copyright © 2012 Society of Chemical Industry     

Effects of high hydrostatic pressure on physicochemical and functional properties of walnut (Juglans regia L.) protein isolate

BACKGROUND: Walnut (Juglans regia L.) is a good source of protein that has potential application in new product formation and fortification. The main objectives of this study were to investigate the effects of high hydrostatic pressure (HHP) treatment (300–600 MPa 20 min) on physicochemical and functional properties of walnut protein isolate (WPI) using various analytical techniques at room temperature. RESULTS: The results showed significant modification of solubility, free sulfhydryl content and surface hydrophobicity with increased levels of HHP treatment, indicating partial denaturation and aggregation of proteins. Differential scanning calorimetry and fluorescence spectrum analyses demonstrated that HHP treatment resulted in gradual unfolding of protein structure. Emulsifying activity index was significantly (P < 0.05) increased after HHP treatment at 400 MPa, but significantly decreased (P < 0.05) relative to the untreated WPI with further increase in pressure. HHP treatment at 300–600 MPa significantly decreased emulsion stability index. Additionally, HHP‐treated walnut proteins showed better foaming properties and in vitro digestibility. CONCLUSION: These results suggest that HHP treatment could be applied to modify the properties of walnut proteins by appropriate of pressure levels, which will help in using walnut protein as a potential food ingredient. © 2012 Society of Chemical Industry     

Anti‐inflammatory properties of high pressure‐assisted extracts of Grifola frondosa in lipopolysaccharide‐activated RAW 264.7 macrophages

The aim of this study was to determine the in vitro anti‐inflammatory properties of the shake extract (SE) and the high pressure‐assisted extract (PE) of the mycelia of Grifola frondosa in a lipopolysaccharide‐stimulated RAW 264.7 macrophage model. The content of total polysaccharides and β‐glucans of PE at 600 MPa (PE‐600) was 41.2 and 6.2 mg g^?1 dry weight, respectively, which were significantly higher than SE extracts. The results showed that treatment with 500 μg mL^?1 of PE by 600 MPa (PE‐600) did not reduce RAW 264.7 cell viability but did significantly inhibit the production of LPS‐induced NO, PGE₂ and intracellular ROS. The PE‐600 inhibited the activation of NF‐kB and then reduced the production of LPS‐induced TNF‐α, IL‐6 and IL‐1β in a dose‐dependent manner. Thus, the PE could be used as an alternative extraction method for improving the extraction efficacy of G. frondosa and serve as an alternative source of anti‐inflammatory agents.     

Microbial and Sensory Effects of Combined High Hydrostatic Pressure and Dense Phase Carbon Dioxide Process on Feijoa Puree

High hydrostatic pressure (HHP) is used for microbial inactivation in foods. Addition of carbon dioxide (CO₂) to HHP can improve microbial and enzyme inactivation. This study investigated microbial effects of combined HHP and CO₂ on Escherichia coli, Bacillus subtilis, and Saccharomyces cerevisiae, and evaluated sensory attributes of treated feijoa fruit puree (pH 3.2). Microorganisms in their growth media and feijoa puree were treated with HHP alone (HHP), or saturated with CO₂ at 1 atm (HHPcarb), or 0.4%w/w of CO₂ was injected into the package (HHPcarb+CO₂). Microbial samples were processed at 200 to 400 MPa, 25 °C, 2 to 6 min. Feijoa samples were processed at 600 MPa, 20 °C, 5 min, then served with and without added sucrose (10%w/w). Treated samples were analyzed for microbial viability and sensory evaluation. Addition of CO₂ enhanced microbial inactivation of HHP from 1.7‐log to 4.3‐log reduction in E. coli at 400 MPa, 4 min, and reduction of >6.5 logs in B. subtilis (vegetative cells) starting at 200 MPa, 2 min. For yeast, HHPcarb+CO₂ increased the inactivation of HHP from 4.7‐log to 6.2‐log reduction at 250 MPa, 4 min. The synergistic effect of CO₂ with HHP increased with increasing time and pressure. HHPcarb+CO₂ treatment did not alter the appearance and color, while affecting the texture and flavor of unsweetened feijoa samples. There were no differences in sensory attributes and preferences between HHPcarb+CO₂ and fresh sweetened products. Addition of CO₂ in HHP treatment can reduce process pressure and time, and better preserve product quality.     

Antibacterial activity of Ginseng (Panax ginseng C. A. Meyer) stems–leaves extract produced by subcritical water extraction

Antibacterial activities of ginseng extracts produced from ginseng by‐products, stems and leaves, using subcritical water extraction (SWE) at 110, 165 and 190 °C, were evaluated and compared with those of ginseng extracts prepared by hot water and ethanol extraction. The ginseng stems–leaves extract produced by SWE at 190 °C contained the greatest concentration of phenolics (98.4 mg GAE g^?1 of extract). All ginseng extracts inhibited the growth of Bacillus cereus, Salmonella enteritidis, Escherichia coli O157:H7 and Listeria monocytogenes. Among four strains, B. cereus was more sensitive to the ginseng extract by SWE at 190 °C than other bacteria. Cell membranes of bacteria were disrupted by the addition of SWE ginseng extract, observed by transmission electron microscopy (TEM), with the release of cellular contents. These findings provided evidence about the potential utilisation of ginseng stems and leaves by using an environmental friendly extraction process, SWE, to produce ginseng extract for the inhibition of bacteria growth.     

Anthocyanin extracts from purple sweet potato by means of microwave baking and acidified electrolysed water and their antioxidation in vitro

Two anthocyanin extracts from purple sweet potato (PSP) were prepared by means of microwave baking (MB) and acidified electrolysed water (AEW) or 95% ethanol. The extraction yield in AEW (pH 3.0) was up to 35.0% nearly 2.5 times higher than in ethanol. When pH ≤ 3.0, the lower the pH values of the extracts in solution were, the darker the red extracts were. Total flavonoids, phenolic and monomeric anthocyanin contents in AEW extract were 132.13, 64.52 and 102.31 mg g^?1, respectively, whose values were the similar to or slightly lower than those in ethanol extract. On the contrary, its total sugar content (61.31 mg g^?1) was nearly five times higher than that of the ethanol extract. In vitro assay indicated that the scavenging capability of DPPH free radicals of AEW extracts (IC₅₀ = 12.0 μg mL^?1) was stronger than that of the ethanolic ones. The reducing power and inhibiting lipid peroxidation of the two extracts were similar. Thus, the new extraction of MB‐AEW described here was not only simple and low in cost, but also had much higher extraction yield. The anthocyanin extracts with a strong antioxidation and a stable red colour could be widely used as food colouring additives and anti‐ageing health foods.     

Effect of processing methods on the concentrations of bioactive components of ginseng (Panax ginseng C.A. Meyer) adventitious roots

Adventitious roots of Korean mountain ginseng (Panax ginseng C.A. Meyer) were processed by using forced air drying methodology at 30, 50 and 70 °C for 1, 3, 5, 10 and 20 h with objective of developing suitable processing/drying technique. Drying of adventitious roots at 50 °C for 10 h was found suitable as desirable moisture content (3.13 g water/dry matter i.e. 10%) could be reached with dried roots. Roots which were dried by such treatment were also possessing higher amount of ginsenosides (1.5 mg g⁻¹ DW triols, 15.9 mg g⁻¹ DW diols and 17.4 mg g⁻¹ DW of total ginsenosides) and phenolics. Adventitious roots were also processed by using far infrared and freeze drying methods and results revealed that forced air drying method is superior to far infrared and freeze drying methods.     

Detection of Panax quinquefolius in Panax ginseng using 'subtracted diversity array'

BACKGROUND: Food adulteration remains a major global concern. DNA fingerprinting has several advantages over chemical and morphological identification techniques. DNA microarray‐based fingerprinting techniques have not been used previously to detect adulteration involving dried commercial samples of closely related species. Here we report amplification of low‐level DNA obtained from dried commercial ginseng samples using the Qiagen^? REPLI‐g^® Kit. Further, we used a subtracted diversity array (SDA) to fingerprint the two ginseng species, Panax ginseng and Panax quinquefolius, that are frequently mixed for adulteration. RESULTS: The two ginseng species were successfully discriminated using SDA. Further, SDA was sensitive enough to detect a deliberate adulteration of 10% P. quinquefolius in P. ginseng. Thirty‐nine species‐specific features including 30 P. ginseng‐specific and nine P. quinquefolius‐specific were obtained. This resulted in a feature polymorphism rate of 10.5% from the 376 features used for fingerprinting the two ginseng species. The functional characterization of 14 Panax species‐specific features by sequencing revealed one putative ATP synthase, six putative uncharacterized proteins, and two retroelements to be different in these two species. CONCLUSION: SDA can be employed to detect adulterations in a broad range of plant samples. Copyright © 2011 Society of Chemical Industry     

Effect of high hydrostatic pressure on the structure,physicochemical and functional properties of protein isolates from cumin (Cuminum cyminum) seeds

The effect of high hydrostatic pressure (HHP) treatment on the structure, physicochemical and functional properties of cumin protein isolate (CPI) was investigated. More aggregates, pores, irregular conformations and bigger particle size were observed for HHP-treated CPI. HHP resulted in an increase in α-helix, a decrease in β-strand and fluorescence intensity of CPI. Surface hydrophobicity (H_o) of CPI significantly increased after HHP treatment, from 343.35 for native CPI to 906.22 at 600 MPa (P < 0.05). HHP treatment at 200 MPa reduced zeta-potential and solubility of CPI, while had little effect at 400 and 600 MPa. Emulsifying activity and stability of CPI decreased after HHP treatment, of which droplet size of emulsions significantly increased (P < 0.05). HHP-treated CPI could form heat-induced gelation at lower temperature (68.5 °C) and improved storage modulus (G′) comparing to native one (80.6 °C), suggesting that CPI might be potential protein resources as gelation substitute in food system.     

Effect of nitrapyrin on nitrapyrin residues and nitrate content in red beet roots fertilised with urea

The effect of nitrapyrin [2-chloro-6-(trichloromethyl)pyridine] application on nitrapyrin residues and nitrate content in red beet (Beta vulgaris L. var. conditiva) roots and in soil fertilised with urea was studied during the 1977 growing season. The nitrapyrin residues in roots were on 15 August between 0.00–1.19 μg g^?1 and on 1 October between 0.00–0.45 μg g^?1 (dry weight basis) correlating significantly r = 0.99, P<0.01) to the rates of nitrapyrin applied on 8 June (0–34 kg ha^?1). The content of nitrapyrin in roots decreased in autumn having in August-September a ‘biological half-life’ of 3 weeks. In soil the same parameter was in July-September more than 4 months. The ratio of nitrapyrin to its main breakdown product varied in soil between 3.5–24.8 and in red beets between 0.6–1.4. Neither of these compounds were found in processed canned red beets. Nitrapyrin treatment decreased the nitrate content in both soil and beets. The inhibitor application of 18 kg ha^?1 caused the highest reduction of the NO₃-nitrogen content in beets, the average values on dry weight basis being 0.51–0.34 mg g^?1 for 0–18 kg ha^?1 nitrapyrin application, respectively. The reduction of NO₃-nitrogen in beets from its level of 15 August to its September level was 44%. After 5 September the reduction of the nitrate level in beets was not significant.     

A comparative study of changes in the microbiota of apple juice treated by high hydrostatic pressure (HHP) or high pressure homogenisation (HPH)

The objective of this study was to assess the effect of High Pressure Homogenisation (HPH) compared with High Hydrostatic Pressure (HHP) on the microbiological quality of raw apple juice during storage at ideal (4 °C) and abuse (12 °C) temperatures. In the case of HPH, only low numbers of micro-organisms were detected after treatment at 300 MPa (typically between 2 and 3 log.ml⁻¹). These were identified as Streptomyces spp., and numbers did not increase during storage of the juice for 35 days, irrespective of storage temperature. In the case of HHP, the total aerobic counts were also reduced significantly (p < 0.05) after treatment for 1 min at 500 and 600 MPa and the numbers did not increase significantly during storage at 4 °C. However, during storage at 12 °C the counts did increase significantly (p < 0.05) and by day 14 counts at 500 MPa were not significantly different from the control juice. This confirms that good temperature control is important if the full benefits of HHP treatment are to be realised.Frateuria aurantia dominated the microbiota of the HHP apple juice stored at 12 °C along with low levels of Bacillus and Streptomyces spp.The HPH and HHP juices both turned brown during storage indicating that neither treatment was sufficient to inactivate polyphenol oxidase. The enzyme is known to be pressure resistant and this discolouration was controlled by a heat treatment (70 °C for 1 min) used in commercial practice and given prior to HP treatment.