Journal of the Science of Food and Agriculture

J Sci Food Agric 79 :663670 (1999)

Effects of microwave heating on pigment composition and colour of fruit purees

Begona de Ancos , M Pilar Cano,* Almudena Hernandez and Marianne 8 Monreal
Plant Foods Science & Technology Department , Ins tituto del Fr o (CSIC ), Ciudad Univers itaria , 28040 -Madrid , Spain

Abstract : Microwave energy was applied to inactivate the oxydoreductases peroxidase (POD, EC 1.11.1.7) and polyphenol oxidase (PPO, EC 1.14.18.1) in processed fruit products. Microwave blanching of papaya, strawberry and kiwi purees at various conditions of power and time produced inactivation of PPO and POD activities depending on the fruit and the heating conditions. Treatment at 850 W/60 s produced about 60% of POD inactivation for papaya and kiwi fruit. POD activity in strawberry, however, seemed to be more resistant to microwave inactivation ; treatment at 850 W/60 s only achieved a loss of POD activity near 8% . Papaya oxidoreductases showed lower stability in the microwave treatments tested. Microwave blanching at 475 W/45 s produced about 75% inactivation of POD activity and nearly complete PPO inactivation. Kiwi fruit and strawberry purees exhibited similar inactivation of PPO 32% at 475 W/30 s and 70% at 475 W/60 s. The decrease of PPO activity in both products was almost linear at constant power. This thermal treatment, however, directly a ects the colour of the fruit pulps. Papaya, kiwi and strawberry purees su ered slight colour (CIE L*a*b*) changes. Carotenoid, chlorophyll and anthocyanin changes were evaluated by HPLC and related to objective colour. Microwave treatments produced small modications of the quantitative and qualitative composition of carotenoids (in papaya) and anthocyanins (in strawberry). Chlorophylls (kiwi) showed signicant degradation as a consequence of microwave heating. ( 1999 Society of Chemical Industry

Keywords : microwave ; polyphenol oxidase ; peroxidase ; pigments ; kiwi fruit ; papaya; strawberry ; purees ; colour INTRODUCTION Variability in colour and lack of colour stability are major problems experienced with processed fruit products. Undesirable sensory and biochemical changes during handling, processing and storage of fruit products are a result of enzymatic browning1 or of non-enzymatic reactions (Maillard mechanisms).2 Development of browning (or discoloration), oavours and nutritional damage were attributed to the action of enzymes polyphenol oxidase (PPO, EC 1.14.18.1) and peroxidase (POD, EC 1.11.1.7).3h6 The use of microwave energy to inactivate enzymes prior to processing fruits and vegetables is not a common practice. The advantages of the use of microwave energy when compared with conventional heat-blanching are : (1) in-depth heating in the absence of a temperature gradient ; (2) inactivation of enzyme complexes and (3) avoidance of the leaching of vitamins, avours, pigments, carbohydrates and other water-soluble components.
This work was partially pres ented as a pos ter to the IX World Congres s of Food Science & Technology, held in Budapes t (Hungary) 31 July4 Augus t, 1995 * Corres pondence to : M Pilar Cano, Plant Foods Science & Technology Department, Ins tituto del Fri o (CSIC), Ciudad Univers itaria, 28040-Madrid, Spain

Microwave blanching operations have been applied successfully in the enzymatic inactivation in whole tomato fruits and whole soybeans.7,8 Eects of water- and microwave-blanching methods on activities of peroxidases and lipoxygenases in green beans, peas and carrots were reported by Guenes and Bayindirli,9 who concluded that the less severe heat treatment required to inactivate the enzymes, when the microwave treatment was applied, should result in improved product avour, colour, texture and nutritional value. A preliminary study of microwave blanching of fruit tissues was made by Cano et al10 in banana products. In this work the eects of steam and microwave blanching on PPO and POD enzymes were described. More recently Cano11 reported that the optimal quality in frozen banana was obtained with microwave pretreatment. Also, Giami12 reported benecial eects of microwave treatment applied prior to freezing plantain (Musa paradisiaca) slices in
Contract/grant s pons or : Comis ion Interminis terial de Ciercia y Tecnologia Contract/grant number : ALI 94-1442, ALI 95-0105 (Received 9 June 1997 ; revis ed vers ion received 5 February 1998 ; accepted 12 Augus t 1998 )

( 1999 Society of Chemical Industry. J Sci Food Agric 0022-5142/99/$17.50

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B de Ancos et al

that the microwaved product had a texture similar to that obtained with calcium chloride treatment. Application of microwave energy in the enzyme inactivation of tropical fruit pulps (guava, papaya and mango) were reported by Abd-El-Al-Mg et al ;13 they found that the exposure times required for enzyme inactivation in pulps varied from 20 to 80 s depending on enzyme and pulp type. There is, however, little information about the application of microwave blanching to other fruits. Colour and pigment degradation in strawberry products have been well documented.14h16 Heat processing and subsequent storage of fruits in whole, sliced, pureed and concentrated form have been accompanied by discoloration due to the degradation of the anthocyanins. Wrolstad et al17 studied the eect of microwave blanching on the colour and composition of strawberry concentrate and juice ; they found that this treatment protected pigment composition and improved colour stability. The most important sensory and nutritional changes that aect kiwi fruit pulp during processing are losses of bright green colour and ascorbic acid content. Cano et al18 studied changes in the major pigment constituents of frozen kiwi fruit slices during prolonged storage at [ 18C and the degradation of chlorophylls and formation of pheophytins and metallochlorphyll complexes (Zn-pheophytin a and Zn-pyropheophytin a) in canned products due to thermal treatments,19 but information about microwave treatment on kiwi fruit products was not reported. The major purpose of this study is to investigate the eect of microwave energy on the PPO and POD activities of strawberry, papaya and kiwi fruit purees, and to study the eects of microwave blanching on the CIE L*a*b* colour values of processed fruit products. In addition, the HPLC analysis of pigment composition of the fruit purees after treatments is undertaken in order to compare heat pretreatment with other traditional methods for the preservation of fruit purees.

Hayward) were purchased from local supermarkets. Ten (kiwi and papaya) and 50 (strawberry) fruits were peeled and inedible parts removed. Fruit purees were obtained by homogenisation using a blender (Osterizer, Proctor-Siles, Inc, North Caroline, USA). Table 1 shows the initial characteristics of these fruit purees. Microwave treatment Samples (50 g) of each fruit puree were placed in 10 cm diameter glass dishes to a depth of 2 cm. Three sample dishes (50 g) were heated in a 850 W household microwave oven (Toshiba ER-6860w, J apan). The power generator for this unit operates at a frequency of 2450 MHz. Treatments applied to the fruit puree were adjustable to the following values : 285 W, 570 W and 850 W for constant time (30 s), or constant power (475 W) for 15 s, 30 s, 45 s and 60 s. After treatment, the purees were placed in a suitable vessel and cooled in ice tap water for 5 min and analyses were carried out immediately. All experiments were carried out in triplicate. Determination of colour data Colour of treated fruit purees was measured in a cylindrical sample cup, 5 cm diameter ] 2 cm high lled to the top, using the Tristimulus reectance colorimeter model HunterLab, model D25 (Hunter Associates Laboratory mod. 25-9, Reston, VA, USA) calibrated with a white tile No. C2-19913 (X \ 82.51, Y \ 84.46, Z \ 101.44). Colour was measured using the CIE L*a*b* colour values, where L* value is a measure of lightness from completely opaque (0) to completely transparent (100), a* is a measure of redness ([a* greenness) and b* of yellowness ([b* blueness). Hue angle (h) was calculated from h \ arctan b*/a*() and chroma (C) from C \ [(a*)2 ] (b*)2]0.5. All measurements were done in triplicate (three dierent dishes). Biochemical analysis The enzyme extracts for determination of POD and PPO were made by homogenisation of 10 g of fresh weight of each fruit puree sample with 20 ml (or 10 ml for kiwi fruit samples) of 0.2 M sodium phosphate buer (pH 7.0 for papaya and kiwi fruit or pH 6.5 for strawberry) containing 40 g litre~1 insoluble polyvinylpolypyrolidone (PVPP) (or 10 g litre~1 for kiwi fruit) and 10 ml litre~1 Triton X-100, in ultra-

MATERIALS AND METHODS Fruit purees Strawberries (Fragaria ananassa ] Duchesne, cultivar Chandler), papayas (Carica papaya, cultivar Sunrise) and kiwi fruits (Actinida chinensis, Planch, cultivar
Table 1. Initial characteris tics of fruits purees a

Characteris tic
Mois ture (g kg1 FW) Total acidity (g anhydrous citric acid kg1 FW) pH Soluble s olids (Brix)

Kiwi
810.00 ^ 0.10 14.50 ^ 0.12 3.39 ^ 0.16 17.10 ^ 0.23

Strawberry
925.50 ^ 0.15 9.40 ^ 0.18 3.40 ^ 0.22 7.73 ^ 0.51

Papaya
859.60 ^ 0.17 1.00 ^ 0.14 6.41 ^ 0.34 12.80 ^ 0.18

a Values are average of three independent determinations ^ s tandard deviation h. FW fres h weight

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E ect of microwave heating on fruit colour

homogeniser (Omnimixer, mod ES-207, Omni International, Inc, Gainsville, VA, USA) with external cooling, for 3 min with stop intervals each 30 s. The homogenate was centrifuged at 16 000 ] g at 4C for 15 min. The supernatant was ltered through a nylon cloth and the volume of the ltrate measured. POD activity was assayed spectrophotometrically using aliquots (0.025 ml) of enzyme extract and a reaction mixture composed by 2.7 ml of 0.05 M sodium phosphate buer (pH 6.5 for papaya and strawberry and pH 7 for kiwi fruit) with 0.2 ml, 10 g litre~1 p-phenylenediamine as H-donor and 0.1 ml 15 g litre~1 hydrogen peroxide as oxidant. The oxidation of p-phenylenediamine was measured with a double beam spectrophotometer (Perkin Elmer, mod Lambda 15, Bodenseewerk, FRG) at 485 nm and 25C. PPO activity was assayed using aliquots (0.075 ml) of enzyme extract and 3.0 ml of a solution of 0.07 M catechol (papaya and strawberry) or a solution of 0.15 M catechol (kiwi fruit) in 0.05 M sodium phosphate buer (pH 6.5). The reaction was measurement at 420 nm at 25C. HPLC pigment analysis Apparatus A Hewlett-Packard 1050 quaternary solvent delivery unit equipped with a Hewlett-Packard 1040A rapid scanning UV-visible photodiode array detector was employed. The data were stored and processed with a Hewlett-Packard Model 9000/300 computing system and Colour Pro-plotter. The absorption spectra of the pigments were recorded between 300 and 600 nm at the rate of 12 spectra min~1. The HP-9000 computer with a built-in integration program was used to evaluate the peak area and peak height. Absorption spectra of isolated components in various solvents were recorded on a Perkin Elmer Lambda 15 UV-visible spectrophotometer (Bodenssewerk, FRG). Separation was performed on a stainless steel (250 ] 4 min id) Hypersil ODS (5 lm spherical particles) column (Hewlett-Packard) protected with guard column cartridge (2 cm length ] 4.0 mm id) packed with ODS-Hypersil C18 (5 lm particle size). Chromatographic procedures The analytical separations of kiwi fruit and papaya pigments were carried out according to the procedure of Cano18 with minor modications for papaya pigments.20 A gradient mixture of methanol/ water (75 : 25 v/v), eluent A, and ethyl acetate, eluent B, was used, beginning at time zero until time 10 min with a penultimate composition of eluent B (70% v). The gradient eluent composition was followed at time 10 min until time 14 min with the nal composition of eluent B (100% v). The ow rate employed was 1.7 ml min~1 and the chromatographic runs were monitored at 430 nm (450 nm for carotenoids). At the end of the gradient the column was re-equilibrated
J Sci Food Agric 79 :663670 (1999)

under the initial conditions by new gradient conditions beginning at time 14 min until time 20 min with a nal composition of eluent B (0% v) at the same ow rate (1.7 ml min~1). Chromatographic conditions for anthocyanin analysis were according to the procedure described by Hong and Wrolstad.21 Solvent A was 4 g litre~1 phosphoric acid and solvent B was 100% acetonitrile and the program began with isocratic elution with 6% v B from 0 to 10 min, and then a linear gradient to 20% v of B from 10 to 55 min, and nally an isocratic elution at 20% v of B from 55 to 65 min. Flow rate employed was 1.0 ml min~1 and the runs were monitored at 520 nm. Extraction of kiwi fruit pigments The general method for extraction was described in a previous study of kiwi fruit pigments.18 The procedure consists of a chilled acetone extraction (200 ml) of 50 g kiwi fruit puree (adjusted to pH 89 with 12 g of sodium carbonate to prevent conversion of chlorophyll to pheophytin). After mixing in a homogeniser (Omnimixer, mod ES-207, Onmi International, Inc, Gainsville, VA, USA) and centrifuging at 4000 ] g for 10 min (05C), the supernatants were collected and transferred to a separatory funnel and diethyl ether (75 ml) and cold deionised water (100 ml) added. After vigorous shaking and standing, the aqueous layer was discarded. The washing procedure was repeated ve times to remove acetone. The diethyl ether layer was dehydrated (anhydrous sodium sulphate). The extracts were evaporated under a stream of N and the residue dissolved in 2 5 ml of chromatographic grade acetone. Duplicate 20 ml samples of each extract were injected for HPLC analysis. The separation and identication of pigments were based on the chromatographic behaviour with HPLC and TLC, visible absorption spectra and specic chemical reactions, as reported previously.18 The pigments were quantied in a given sample by means of a calibration curve that included all of the chlorophylls and carotenoids to be assessed in that sample using the HP-9000 computer system. The curves were prepared daily by diluting portions of the starting solutions to the appropriate proportions for the samples being analysed. Extraction of strawberry pigments Sample preparation and HPLC separation and identication of anthocyanins was carried out according to the procedure described by Hong and Wrolstad.21 Strawberry puree samples (10 g) were homogenised with 100 ml of 10 g litre~1 HCl in methanol. The slurry was ltered, and the solids were washed with an additional 100 ml of 10 g litre~1 HCl in methanol. The methanol extracts were combined and concentrated to about 10 ml in a rotary evaporator (30C). The aqueous extract was placed in a volumetric ask and made up to 50 ml with 0.1 g litre~1 HCl solution. The aqueous solution of anthocyanins (5 ml) was
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adsorbed onto an activated C18 Sep-pack cartridge (Water Associates, Milford, MA)(activated with 3 ml of methanol and 3 ml of 0.01 M HCl solution). The pigments adsorbed onto the cartridge were eluted with 0.1 g litre~1 HCl in HPLC grade methanol. The solution was evaporated to dryness in a rotary evaporator and the extractor was dissolved in 40 g litre~1 phosphoric acid and ltered through a 0.45 mm lter and 20 ll injected into the HPLC system. All the analyses were performed in duplicate. The separation of anthocyanins was carried out by HPLC and the identication was performed by the analysis of spectral properties by an on-line photodiode-array detector. Total pigment concentration was assessed using pelargonidin-3-glucoside, previously separated, as an external standard. A standard curve was prepared by plotting dierent concentrations of pelargonidin-3glucoside versus area measurement in HPLC. Extraction of papaya pigments The extraction was carried out following the procedure described by Cano et al.20 A mix of fruit sample (30 g) and sodium sulphate and magnesium carbonate (200% and 10% of the weight of fruit, respectively), and tetrahydrofuran (THF) (100 ml stabilised with BHT) were homogenised at 0C, in total darkness and under nitrogen atmosphere. After a rigorous clean up, the extracts were evaporated under nitrogen and the residue dissolved in 0.3 ml of dichloromethane. Duplicate 25 ll samples were injected for HPLC analysis. The separation and identication were carried out following the methodology previously reported19 in kiwi fruit. The carotenoids were separated by reversed-phased HPLC and peak purity was determined by electronic spectra using the HP-9000 computer system. Carotenoids were identied according to their chromatographic behaviour on HPLC, TLC and UV-visible absorption spectra, by comparing both their retention time and the absorption spectra obtained with those of authentic carotenoids previously purchased or separated from fruits.18 Pigments functional group examination was carried out by specic chemical reactions. The carotenoids separated were quantied by HPLC using Sudan 1 as internal standard and calculated as b-carotene equivalent using a response factor of 0.0791, in a procedure similar to that of Philip and Chen.22 The response factor, f, was determined by injecting known mixtures of b-carotene and Sudan 1 into the HPLC and measuring the integrator area responses at 450 nm ( f \ conc Sudan 1 ] area b-carotene/area Sudan 1 ] conc b-carotene). Data analysis Statistical analyses were performed using the software InStatTM. Data were analysed statistically by analysis of variance (ANOVA) and using Students t-test.
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RESULTS AND DISCUSSION Effect of microwave treatment on peroxidase activity Figs 1 and 2 show the residual POD activity obtained from microwave treated fruit puree. Microwave heating for 30 s produced almost linear inactivation of POD in kiwi fruit puree. In this product, application of a microwave treatment at 850 W produced near to 60% inactivation of POD if the treatment was performed during 30 s. POD activity in strawberry, however, seemed to be more resistant to microwave inactivation and only a heat treatment at 850 W/30 s rendered a signicant (p 0.05) loss of POD activity ( B 8%), Fig 1. Papaya puree also exhibited a signicant loss of POD activity when a microwave treatment at 570 W/30 s was applied, but no better results were obtained by increasing the power output to 850 W. When the microwave treatments were applied at xed power (475 W) with various times of exposure, similar results could be obtained for POD activity (Fig 2). Papaya POD activity was most eectively inactivated when treatment time was increased. Application of microwaves at 475 W/45 s produced a B 75% inactivation of peroxidase activity and,

Figure 1. Effect of microwave treatments at fixed time (30 s ) on peroxidas e (POD) activity of fruit purees .

Figure 2. Effect of microwave treatments at fixed power (475 W) on peroxidas e (POD) of fruit purees .

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E ect of microwave heating on fruit colour

beyond this exposure time, no signicant increase of enzyme inactivation was observed. Peroxidase activity in kiwi fruit puree was eectively lost by microwave treatments at 475 W from 45 s of exposure time. Prolonged treatments (60 s) did not increase the POD inactivation. Effects of microwave treatments on polyphenol oxidase activity PPO activity changes in microwaved fruit purees are shown in Figs 3 and 4. PPO activity of papaya puree was most eectively inactivated by microwave heating. Power of 570 W for 30 s was enough to produce a 90% loss of PPO activity. This result was similar to that for POD activity in this product (Fig 1), but an evident and greater microwave resistance of peroxidase could be observed when the same conditions of treatment were applied. Kiwi fruit PPO activity was most resistant to microwave heating. In this product, a slight decrease of PPO activity could be obtained (18%, 850 W/30 s, Fig 3), but POD activity in kiwi fruit puree was efficiently inactivated by treatments from 570 W/30 s. Strawberry PPO showed a 50% loss of activity using a mild microwave treatment (285 W/30 s), but

this eectiveness was not maintained when the power was raised to 570 W/30 s. This fact cannot be easily explained by a simple protein denaturation mechanism ; other eects such as thermal rupture of cell organules, which liberates enzyme fractions linked to celular membranes, could result in this higher value of residual PPO activity in strawberry puree treated at intermediate power. Fig 4 represents the residual PPO activity of microwave treated purees at prexed power (475 W). Papaya PPO appeared to be the most labile in these conditions. This product lost all PPO activity by treatment at 475 W for 45 s or more. Again, the lower stability of oxidoreductases, PPO and POD, was found in papaya tissues. Kiwi fruit and strawberry purees exhibited the same inactivation of PPO, B 32% at 475 W/30 s, and B 70% at 475 W/60 s. The reduction of PPO activity in these products by microwave heating was almost linear in these experiments, indicating that the process was better controlled by pre-xing the power than the time of exposure. Effects of microwave treatment on fruit pigment composition Changes of total pigment concentration during microwave blanching are shown in Table 2. The carotenoid composition of Spanish papaya (cv Sunrise) had previously been studied.20 The major carotenoids found in papaya extracts were lycopene and the fatty acid esters of b-cryptoxanthin and bcryptoxanthin-5,6-epoxide. Qualitative carotenoid composition analysed by HPLC was unchanged after microwave treatment, however, microwave treatment induced the degradative loss of total carotenoid content (Table 2). Treatment at 475 W for 45 s showed the greatest loss of the total carotenoid (57%) in this fruit puree. In contrast, during microwave heating there was no signicant change in total anthocyanin concentration (Table 2); the anthocyanin pattern of strawberry was almost unchanged by microwave heating. Other authors had reported an increase of the total anthocyanin contents in microwaved strawberry products compared with controls.17 They attributed this phenomenon to enzymatic action during the process. The increase in anthocyanins content observed in some microwaved treated samples could be explained by a more efficient extraction of these compounds from the tissue due to cellular disruption produced by microwave heating. Microwave heating of kiwi purees produced a signicant decrease of chlorophyll a and b concentration due to chlorophyll degradation through transformation to chlorophyllide a, pheophorbide a and pheophytin b (Table 2). The more drastic treatments, 850 W for 30 s and 474 W for 60 s, tested in our work produced an almost total loss of chlorophyll b, which is the least heat-stable chlorophyll. The kiwi puree blanched at 475 W for 60 s showed
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Figure 3. Effect of microwave treatments at fixed time (30 s ) on polyphenol oxidas e (PPO) activity of fruit purees .

Figure 4. Effect of microwave treatments at fixed power (475 W) on polyphenol oxidas e (PPO) activity of fruit purees .

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Table 2. Effects of microwave treatment on fruit puree pigment concentration (mg kg1 puree)

Pigments * Control 285 W

Papaya Total carotenoids Strawberry Total anthocyanin Kiwi Chlorophyll a Chlorophyll b Xanthophylls Pheophytin a b-carotene Chlorophyllida a Pheophorbide a Pheophytin b
10.5 ^ 0.2a 297 ^ 0.5a 14.0 ^ 0.3a 7.6 ^ 0.2a 6.5 ^ 0.3a 4.8 ^ 0.2a 0.4 ^ 0.1a nd nd nd 7.4 ^ 0.3a 282 ^ 0.3a 3.5 ^ 0.2b 1.1 ^ 0.1b 5.6 ^ 0.4a 3.1 ^ 0.2a 0.2 ^ 0.3a 0.1 ^ 0.4a 0.1 ^ 0.5a nd

Microwave treatment t \ 30 s 570 W

10.8 ^ 0.8a 301 ^ 0.5a 10.3 ^ 0.5c 1.7 ^ 0.1b 4.1 ^ 0.7a 2.7 ^ 0.5a 0.3 ^ 0.4a 0.3 ^ 1.1a 0.1 ^ 0.9a nd

P \ 475 W 850 W
6.0 ^ 1.2a 303 ^ 0.5a 13.6 ^ 0.3a 0.1 ^ 0.2c 6.1 ^ 0.4a 3.1 ^ 0.2a 0.2 ^ 0.4a 0.5 ^ 0.4a 0.2 ^ 0.2a nd

15 s
8.7 ^ 0.4a 321 ^ 0.6a 8.9 ^ 0.4c 0.8 ^ 0.3b 3.9 ^ 0.6ab 2.7 ^ 0.4a 0.1 ^ 0.6a 1.0 ^ 0.6a 0.3 ^ 0.1a nd

30 s
7.4 ^ 0.5a 312 ^ 0.4a 4.0 ^ 0.4b 2.8 ^ 0.3d 2.5 ^ 0.4b 1.2 ^ 0.7ab 0.1 ^ 0.4a 0.4 ^ 0.3a 0.2 ^ 0.5a 0.3 ^ 0.2a

45 s
4.5 ^ 0.6a 332 ^ 0.6a 5.9 ^ 0.1b 1.1 ^ 0.2b 2.0 ^ 0.2b 0.3 ^ 0.5b 0.1 ^ 0.5a 0.2 ^ 0.3a 0.1 ^ 0.2a 0.2 ^ 0.4a

60 s
8.9 ^ 0.6a 314 ^ 0.2a 5.2 ^ 0.5b 0.5 ^ 0.4b 1.8 ^ 0.1b 0.3 ^ 0.4a 0.1 ^ 0.5a 0.5 ^ 0.4a nd 0.15 ^ 0.6a

* Means and s tandard deviations of triplicate determinations . Values in the s ame row with the s ame letter are not s tatis tically different (p \ 0.05) Concentration calculated by HPLC as b-carotene equivalents Concentration calculated by HPLC as pelargonidin-3-glucos ide equivalents

the most notable pigment losses. This sample suffered losses of 72% of xanthophylls, 84% of chlorophyll a and 75% of b-carotene. Coincident with the thermal degradation of pigments, other mechanisms of chlorophyll modication could have occurred in the kiwi fruit purees, such as enzymatic reactions that result in bleaching of fruit tissue. In fact, kiwi puree blanching at 475 W for 60 s retains approximately 60% of the peroxidase activity and B 35% of the polyphenol oxidase activity compared with the control. Effect of microwave treatment on fruit puree objective colour Eects of microwave blanching on fruit puree objective colour are shown in Tables 3, 4 and 5. Micro-

wave blanching at 285 W for 30 s produced the lowest total colour dierence (*E*) in the three fruit purees. Total colour dierence parameter combines L* (luminosity), a* (greenness-redness) and b* (blueness-yellowness) parameters by integrating these three characteristics in order to compare the colour of control and puree samples microwaved at dierent conditions. The *E* value increases when microwave power increases. Microwave blanching did not signicantly aect the papaya CIE L*a*b* colour values (Table 3). Only a minor decrease of a* value and slight increase of b* value were observed, showing that papaya treated purees tend to be less red and slightly more yellow than untreated ones. Total colour dierence (*E*) calculated for microwaved strawberry puree showed signicant dier-

Table 3. Effects of microwave treatment on papaya puree colour (CIE) meas urements

Colour values * Control 285 W L* a* b* Hue angle Chroma *E *

32.30a 13.52a 37.33a 70.09a 39.70a 33.43a 13.81a 39.42a 70.69a 41.77a 2.39a

Microwave treatment t \ 30 s 570 W

35.04a 12.68a 41.10a 72.85a 43.01b 4.73b

P \ 475 W 850 W
33.43a 11.16a 38.58a 73.87a 40.16a 2.90a

15 s
33.77a 12.61a 40.01a 72.51a 41.95a 3.18a

30 s
33.99a 11.27a 39.57a 74.10a 41.14a 3.59a

45 s
33.65a 12.13a 37.58a 72.11a 39.49a 1.95a

60 s
32.31a 10.97a 35.01a 72.60a 36.69a 3.45a

* Values in the s ame row with the s ame letter are not s tatis tically different (p 0.05) h \ arctan (b */a *) (hue angle). *E * \ [(L * [ L *)2 ] (a * [ a *)2 ] (b * [ b *)2]1@2 (total colour difference) 1 2 1 2 1 2 Chroma \ (a *2 ] b *2)1@2

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Table 4. Effects of microwave treatment on s trawberry puree colour (CIE) meas urements

Colour values * Control 285 W L* a* b* Hue angle Chroma *E

30.15a 29.13a 21.20a 36.02a 26.13a 30.77a 29.70a 22.08a 36.63a 27.87a 1.26a

Microwave treatment t \ 30 s 570 W

32.77a 29.28a 22.07a 37.01a 27.34a 2.84a

P \ 475 W 850 W
35.31b 28.89a 21.86a 37.11a 26.54a 5.22b

15 s
33.46a 28.70a 20.17a 35.06a 27.69a 3.73b

30 s
33.50a 29.81a 21.89a 36.28a 29.25b 3.48b

45 s
36.05b 29.48a 22.35a 37.17a 27.92a 6.03b

60 s
36.45b 29.21a 21.05a 35.79a 26.99a 6.30b

* Values in the s ame row with the s ame letter are not s tatis tically different (p 0.05) h \ arctan (b */a *) (hue angle). *E * \ [(L * [ L *) ] (a * [ a *) ] (b * [ b *)]1@2 (total colour difference) 1 2 1 2 1 2 Chroma \ (a *2 ] b *2)1@2

ences as the severity of heating increased (Table 4). There were no signicant dierences between a* and b* values of treated strawberry puree compared with the untreated one. L* values ranged from 30.15 for the untreated sample to 35.31, 36.05 and 36.45 for the more drastically heated samples (850 W/30 s, 475 W/45 s and 475 W/60 s, respectively); the CIE L*a*b* colour values underwent more modication. There were no good correlation between the anthocyanin concentration of the blanched fruit purees and the a* and b* colour values ; although the L* value expresses the darkness of the samples, there was only a moderate correlation between the concentration of anthocyanins and L* value (r2 \ 0.595). The slight browning observing with strawberry puree could be due to slight inactivation of peroxidase and polyphenol oxidase activity during the microwave treatments tested (Table 2). The changes in objective colour parameters of kiwi purees after the various microwaved treatments are showed in Table 5. The total colour dierence (*E*) continuously increased when microwave power increased. This change could be related to the losses of chlorophylls and total xanthophylls during the microwave blanching. Also, the changes in other

objective colour parameters, such as the angle h, a* and b* values, after microwave treatment could explain the losses or breakdown of these two kinds of pigments. The colour parameters a* (greenness) and L* (lightness) increased after almost all the assayed microwaved treatments and this fact was related to the less green and lower lightness of the resultant kiwi purees, when compared with the control. Correlation coefficients of r2 \ 0.8913 (treatment during 30 s/ dierent powers) and r2 \ 0.9620 (treatment at 475 W/ dierent times) were obtained for the ratio chlorophyll a*/ Chroma. The highest correlation between the most important chemical class of pigments in this fruit and CIE L*a*b* colour parameter (Chroma) reected the greater exterior browning of kiwi fruit products compared with the control. CONCLUSION Microwave heating could be an eective treatment to inactivate oxidoreductases enzymes (PPO and POD) in papaya, strawberry and kiwi fruit purees prior to storage or use in various industrial processes. Microwave heating produced small modications of the qualitative and quantitative composition of carotenoids (papaya) and anthocyanins (strawberry)

Table 5. Effects of microwave treatment on kiwi fruit puree colour (CIE) meas urements

Colour values * Control 285 W L* a* b* Hue angle Chroma *E

36.01a [ 12.35a 23.03a [ 61.77a 26.13a 36.39a [ 13.31a 24.48a [ 61.45a 27.88a 1.81a

Microwave treatment t \ 30 s 570 W

37.77a [ 12.85a 24.13a [ 61.96a 27.34a 2.28a

P \ 475 W 850 W
40.07b [ 11.99a 23.68a [ 63.13a 26.54a 4.13b

15 s
40.23b [ 12.19a 24.84a [ 63.75a 27.69a 5.02b

30 s
38.52a [ 11.72a 26.80a [ 66.38b 29.25a 4.60b

45 s
41.04b [ 10.78a 25.76a [ 67.28b 27.93a 5.94b

60 s
40.84b [ 11.17 24.55a [ 65.59b 26.99a 5.32b

* Values in the s ame row with the s ame letter are not s tatis tically different (p 0.05) h \ arctan (b */a *) (hue angle). *E * \ [(L * [ L *)2 ] (a * [ a *)2 ] (b * [ b *)2]1@2 (total colour difference) 1 2 1 2 1 2 Chroma \ (a *2 ] b *2)1@2

J Sci Food Agric 79 :663670 (1999)

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without important changes of the original colour of the fruit purees. By selecting the microwave blanching conditions it is possible to produce a moderate degradation of chlorophylls and a slight loss of the bright green colour of the kiwi fruit puree.

ACKNOWLEDGEMENT This research was supported by the Comision Inter ministerial de Ciencia y Tecnolog a through the project numbers ALI94-1442 and ALI95-0105.

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