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Turk J Agric For 27 (2003) 353-360 © TÜB‹TAK

Kinetics of Nonenzymatic Browning Reaction in Citrus Juice Concentrates during Storage

Nuray KOCA, Hande SELEN BURDURLU, Feryal KARADEN‹Z*
Ankara University, Faculty of Agriculture, Department of Food Engineering, D›flkap› 06110 Ankara - TURKEY

Received: 16.05.2003

Abstract: The kinetics of nonenzymatic browning in citrus juice concentrates (orange, lemon, grapefruit and tangerine) during 8 weeks of storage at 28, 37 and 45 ºC were investigated. Browning development was followed by measuring absorbance at 420 nm (A420) and using CIE-Lab color system. Analysis of kinetic data from A420 values suggested a zero-order reaction for nonenzymatic browning, while changes in L* and b* parameters followed a first-order reaction. Activation energy for nonenzymatic browning determined by A420 values ranged from 17.60 to 35.27 kcal mol-1, while those for L* and b* parameters were 6.67-28.99 kcal mol-1 and 15.38-34.2 kcal mol-1, respectively. Activation energies were higher in orange (28.99-35.27 kcal mol-1) and tangerine (27.84-33.1 kcal mol-1) juice concentrates than those in grapefruit (6.74-27.81 kcal mol-1) and lemon (6.67-17.6 kcal mol-1) juice concentrates. The lower activation energies determined for grapefruit and lemon juice concentrates indicated that nonenzymatic browning reactions are favored in these samples. Key Words: Citrus juice concentrate, nonenzymatic browning, color, storage, kinetic

Turunçgil Suyu Konsantrelerinin Depolanmas›nda Enzimatik Olmayan Esmerleflme Reaksiyon Kineti¤i
Özet: 28, 37 ve 45 °C’de 8 haftal›k bir depolama süresince turunçgil suyu konsantrelerinde (portakal, limon, greypfrut ve mandarin) enzimatik olmayan esmerleflme kineti¤i incelenmifltir. Esmerleflme, 420 nm’de absorbans ölçülerek ve CIE-Lab renk sistemi kullan›larak izlenmifltir. L* ve b* parametrelerindeki de¤iflim birinci dereceden bir reaksiyon izlemekteyken, A420 de¤erlerine ait kinetik veri analizi s›f›r›nc› dereceden bir reaksiyonu göstermektedir. A420 de¤erlerinden belirlenen aktivasyon enerjisi 17.6 ile 35.27 kcal mol-1 aras›nda de¤iflmekteyken, L* ve b* parametrelerinden hesaplanan de¤erler s›ras›yla 6.67-28.99 kcal mol-1 ve 15.38-34.2 kcal mol-1 aral›¤›nda de¤iflmektedir. A420, L* ve b* parametrelerinden hesaplanan aktivasyon enerjileri portakal (28.99-35.27 kcal mol-1) ve mandarin (27.84-33.1 kcal mol-1) suyu konsantrelerinde greypfrut (6.74-27.81 kcal mol-1) ve limon (6.67-17.6 kcal mol1 ) suyu konsantrelerininkinden daha yüksek bulunmufltur. Greypfrut ve limon suyu konsantreleri için belirlenen daha düflük aktivasyon enerjileri, bu örneklerde enzimatik olmayan esmerleflme reaksiyonlar›n›n teflvik edildi¤ini göstermektedir. Anahtar Sözcükler: Turunçgil suyu konsantresi, enzimatik olmayan esmerleflme, renk, depolama, kinetik

Introduction Browning reactions in foods are widespread phenomena that take place during processing and storage (Eskin, 1990). These reactions involve caramelization, ascorbic acid degradation and the Maillard reaction (Clegg, 1964). Since nonenzymatic browning affects the sensory characteristics of cooked foods such as flavor, aroma and color (Gazzani et al., 1987), it may be desirable in baked, fried or roasted foods (Ashoor and Zent, 1984) as in the manufacture of coffee, tea, beer,
* Correspondence to: karadeni@agri.ankara.edu.tr

bread and cake (Martins et al., 2001). However, nonenzymatic browning reactions are often responsible for important quality changes that occur during the storage of foods, limiting their shelf-life (Buera et al., 1987). For instance, browning is the most common quality problem of many concentrated fruit juices (Toribio and Lozano, 1984) and causes loss of nutrients and the formation of intermediate undesirable compounds, like furfural and 5-hydroxymethylfurfural (Buedo et al., 2001).

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Absorbance at 420 nm has generally been used by other researchers (Johnson et al. 11.. Furfural is known to undergo polymerization and. grapefruit (59. the presence of amino acids in ascorbate systems is also considered a major contributor to the development of browning (Robertson and Samaniego. Likewise.... Evaluations were based on a P < 0. 1989. 1992. Determination of water soluble brown pigments: Nonenzymatic browning was monitored using the method of Nagy et al. may combine with amino acids and contribute to the browning of the juice (Solomon et al. nonenzymatic browning is due to the reactions of sugars. as an active aldehyde. Rouseff et al.0. grapefruit and tangerine juice concentrates were 1. Osaka. amino acids and ascorbic acid (Johnson et al. However. Japan) equipped with a DP-300 data processor. 37 and 45 ºC for 8 weeks. HMF concentration has a high correlation with the level of browning in lemon juice and therefore plays an important role in the formation of brown pigments (Robertson and Samaniego. (1990) on a Unicam UV-VIS (UV2) spectrophotometer in 10 mm cells against water at 420 nm. L* and b* values were determined by MINITAB (Version 13) statistical software. orange. 10. Color measurement: Browning development was determined using 2 color measurement methods: (i) absorbance at 420 nm. Another goal of this study was to determine the correlation between the data obtained from both color measurement systems. 1986.Kinetics of Nonenzymatic Browning Reaction in Citrus Juice Concentrates during Storage In citrus juices. Tokyo).20. Nagy et al. orange. CIE-Lab Color System: CIE L* (lightness) and b* (yellowness) values were measured with a Minolta CR 300 Colorimeter (Minolta..57 and 3.and first-order kinetic models were Materials and Methods Materials Commercially processed citrus juice concentrates– lemon (44.. ii. Methods The soluble solids content of concentrates was determined as ºBx using a refractometer (N.0) involved (Clegg.. Robertson and Samaniego. Nippon Optical Work Co. the decomposition of ascorbic acid is reported to be the major deteriorative reaction occuring during the storage of orange juice (Solomon et al. 1990.. lemon. 1964). However. CIE color indices have also been used for measuring browning in foods (Lee and Nagy. Nagy et al. 1988. Belgium). The pH of samples was determined with a pH meter (Consort P407.0 and 11. Ltd. 1986. On the other hand.0 is furfural (Huelin et al. 1995. All citrus juice concentrates in glass jars were stored in darkness at 28. Browning in citrus juices has been studied by a number of investigators (Babsky et al. This is illustrated by the fact that the main degradation product of juices with pH values below 4.0 ºBx) and tangerine (59. Lee and Nagy (1988) also reported a high correlation between the percentage loss of ascorbic acid and an increase in browning in grapefruit juices.. The pH values of lemon.0 ºBx). For this purpose.5 ºBx)..23.5 ºBx)– were obtained from a local processing company. 1995). sugar-amino acid reactions of the classsical Maillard type are of minor importance in citrus juice browning because of the high acidity (pH 2. Johnson et al. 1987) to determine the browning rate in citrus juices. and b* values.. Browning was measured weekly with 2 replications.. The main objective of this study was to determine the kinetics of brown color development in citrus juice concentrates by measuring both water soluble brown pigments (A420) and CIE-Lab color indices. respectively. Schott Gerate. (ii) CIE-Lab Color Scale. 1988. 1999)..W. 1995). respectively.83..O. Zero. Kacem et al. 1995).05 significance level.2. orange (61. 2. The most common method for characterizing browning is the measurement of absorbance at 420 nm (A420). 1986). 1995).2. 354 . Statistical analysis: Correlation coefficients between absorbance at 420 nm (A420). Ibarz et al. 3. Results and Discussion Browning development in citrus juice concentrates was determined using 2 color measurement methods: determination of soluble brown pigments as absorbance at 420 nm (A420) and measurement of CIE L*. i. 1971).04.. grapefruit and tangerine juice concentrates were diluted with distilled water to their ºBx levels of reconstituted juice from concentrates reported in Codex (2001) as 8. 1986). Handwerk and Coleman. In addition.

355 .2 2 1. Burdurlu and Karadeniz. Changes in lightness value (L*) of citrus juice concentrates during storage.6 0. which are the best parameters for the determination of browning in citrus juices.2 0 0 1 2 3 4 5 Storage (week) 6 7 8 9 orange 28 °C orange 37 °C orange 45 °C lemon 28 °C lemon 37 °C lemon 45 °C grapefruit 28 °C grapefruit 37 °C grapefruit 45 °C tangerine 28 °C tangerine 37 °C tangerine 45 °C Figure 1.38 1. Nonenzymatic browning was described as a zero-order reaction since regression analysis of A420 values and time gave a linear relationship (Figure 1). L* and b* values for citrus juice concentrates during 8 weeks of storage at 28. 1984). 37 and 45 °C are shown in Figures 1-3. Changes in A420.5 1.68 1. F.62 1. KARADEN‹Z used to evaluate data from A420.N. KOCA. SELEN BURDURLU. and 2. 1999). 2002). L* and b*.4 0.8 Absorbance (A420) 1. The kinetic parameters for A420 are given in Table 1.44 1.8 0. apple juices (Lozano.56 Lightness (L*) 1. The most appropriate model was selected on the basis of determination coefficients (R2) obtained from regression analysis.4 1.. Changes in absorbance at 420 nm for citrus juice concentrates during storage. respectively.2 1 0.. 1. 1991. pear juice concentrate (Beveridge and Harrison. Rate constants were determined from best-fit regression equations. H. pear puree (Ibarz et al. Previous studies on nonenzymatic browning reactions based on A420 measurement in citrus juices (Saguy et al.26 1. 1978).6 1.2 0 1 2 3 4 5 6 Storage (week) 7 8 9 orange 28 °C orange 37 °C orange 45 °C lemon 28 °C lemon 37 °C lemon 45 °C grapefruit 28 °C grapefruit 37 °C grapefruit 45 °C tangerine 28 °C tangerine 37 °C tangerine 45 °C Figure 2.32 1.

9664) 1.9883) Ea (kcal mol-1) 35.9832) 1.35 37-45 ºC 2. Table 1. In addition.0804 (0. Kinetic parameters for water soluble brown pigments (A420) in citrus juice concentratesa. (2001) on the basis of A420.5008 (0. first-order kinetics were used to evaluate these color data.9803) 4. a first-order browning reaction was reported in orange juice serum (Johnson et al.9150) 9.9933) 1. Q10 Species Temperature (ºC) 28 Orange 37 45 28 Lemon 37 45 28 Grapefruit 37 45 28 Tangerine 37 45 a -lnk 4. 1983) also confirmed that the reactions followed zero-order kinetics.9942) 4.88 28-45 ºC 6. On the other hand.6084 (0.28 1.Kinetics of Nonenzymatic Browning Reaction in Citrus Juice Concentrates during Storage 1.46 Numbers in brackets represent the determination coefficients (R2) intermediate moisture apples (Singh et al.9675) 2.4741 (0.71 32.9934) 3. Changes in b* for citrus juice concentrates during storage.39 (0.9224) 13.9 lemon 45 °C grapefruit 28 °C grapefruit 37 °C 0.1312 (0.9803) 2. 1995) and apple juice concentrates (Toribio and Lozano.16 lemon 28 °C lemon 37 °C b* 0..42 orange 37 °C orange 45 °C 1.2586 (0. indicating that the color of samples changed from yellow to brown. 1984).60 (0. L* and b* values decreased as a function of storage time and temperature (Figures 2 and 3).59 27.46 2.68 orange 28 °C 1.64 grapefruit 45 °C tangerine 28 °C 0.9878) 2.68 4.9471) 3.53 1..9784) 1. Since regression analysis of the logarithm of L* and b* values and time gave a higher linear relationship.3591 (0. a parabolic kinetic model of the nonenzymatic browning reaction of peach juice concentrate was determined by Buedo et al.80 2.27 (0.8859 (0. 356 The kinetics of the change in L* and b* values in citrus juice concentrates were also evaluated.4609 (0.26 17.5 5.9328) 28-37 ºC 15.5877 (0. The first-order kinetic model was also applied to L* changes by Lozano and Ibarz .0122 (0.38 tangerine 37 °C tangerine 45 °C 0.0372 (0.12 0 1 2 3 4 5 Storage (week) 6 7 8 9 Figure 3.81 (0.9573) 2.

..5 0 3.6 3..99 kcal mol-1..2 3. Arrhenius plots of L* in citrus juice concentrates.. F..23 3.32 The temperature dependence of browning was determined by calculating activation energies (Ea) and temperature quotients (Q10) by Eqs... As shown in Table 1... and by Ibarz et al.......17 3. Arrhenius plots for A420 values in citrus juice concentrates.....N..20) and tangerine (pH 3.6 4 .4 2....2 4. (1999) for pear puree. Thermal degradation of the color in peach puree using L* and b* parameters was also found to be a first-order reaction by Ávila and Silva (1999)...2 1.5 1 0... SELEN BURDURLU.29 3...38 to 34.67 to 28..17 3.60 to 35...14 3.17 3.2 3. 1 and 2.... k = rate constant (week-1) -1 ko = pre-exponential factor (week ) 5 4... the Ea values for A420 results ranged from 17.....5 3 .26 1/T 103 (°K) orange lemon grapefruit tangerine 3. (1995) in orange juice serum (19-25 kcal mol-1) and by Ibarz et al.5 2 1.. and Arrhenius plots for A420 values are given in Figure 4.59 to 15..23 3.29 3..5 3 2. Arrhenius plots of b* in citrus juice concentrates.. e ...ln k 3.ln k 2.... While the Ea values for the L* parameter of citrus juice concentrates ranged from 6..ln k 5 4.. (1999).. (1) Figure 5..14 3.. those for the b* parameters ranged from 15......5 1 0. Furthermore. 1996).... H.5 4 3. T2 = absolute temperature (°K) Q10 values obtained from A420. 5. .26 orange lemon grapefrui tangerine 3...29 3.. (2) k1 = rate constant at T1 k2 = rate constant at T2 T1..23) juice concentrates than those in grapefruit (pH 2.2 kcal mol-1..5 2 1...14 orange lemon grapefruit tangerine 3..... Activation energies for L* and b* parameters were calculated from the slopes of the Arrhenius plots shown in Figures 5 and 6.. 3.. The Ea values were comparable with those reported by Johnson et al. peach and prune purees.82) juice concentrates. This result indicated 357 . In relation to parameter L*.7-43.5 4 3.32 k = ko.8 2.88..56) and lemon (pH 1. these values are in the range normally expected for nonenzymatic browning reactions (16.27 kcal mol-1 and Q10 values from 1. (1999) in the liquid fraction of pear puree (15 kcal mol-1).....987 cal-1 mol oK) T = absolute temperature (ºK) Q10 = [k2/k1] 10/T2-T1 ... Tables 2 and 3 shows the activation energies (Ea) and Q10 values for L* and b* parameters.3 kcal mol-1 was obtained for the liquid fraction of pear puree by Ibarz et al.5 0 3. KOCA..26 1/T 103 (°K) 3..32 Ea = activation energy (kcal mol-1) R = gas constant (1....23 1/T 103 (°K) Figure 6... L* and b* parameters were higher in orange (pH 3.... –Ea/RT Figure 4. KARADEN‹Z (1997) for thermally treated apple. an activation energy value of 18.0 kcal mol-1) (O’Brien.2 3...

4546 (0.0515 (0.8078 (0. 1990).8711) 28-37 ºC 20.8642) 2.49 37-45 ºC 1.7806) 1.99 6.8544) 2. the 358 development of browning in lemon and grapefruit juice concentrates may be attributed to their higher acidity.7297 (0.9909) 2.9059) 3. Kinetic parameters for lightness value (L*) in citrus juice concentratesa.5801 (0.47 15. On the basis of browning rates.44 28-45 ºC 5.86 Numbers in brackets represent the determination coefficients (R2) Table 3.99 33.9809 (0. having a central role in the browning of citrus juices and concentrates (Eskin.7780) 2.8689) 3.4427 (0.77 (0.20 (0.9915) 1. it is suggested that browning in orange and tangerine juice concentrates is more temperature dependent.0879 (0.99 (0.9706) 1.8271 (0.6754) 1.7328) 2.9017) 3.7995) 6.8621) 12.8447) 28-37 ºC 14.1 6.9890) Ea (kcal mol-1) 34.9806) 1.38 (0.7434) 3.3 5.21 6.6180 (0.7494 (0.74 (0. Thus.9551) Ea (kcal mo-1) 28.2702 (0.5367 (0.4641 (0.8629 (0.41 0.91 37-45 ºC 1. Ea values followed the same trend as Q10 values for all citrus juice concentrates.9596) 1.22 1. Since ascorbic acid.5475 (0.9681) 2.8667 (0.8852) 17.6539) 5..0056 (0.1653 (0.67 (0.94 2.92 1.35 28-45 ºC 4.5929 (0.87 1. Q10 Species Temperature (ºC) 28 Orange 37 45 28 Lemon 37 45 28 Grapefruit 37 45 28 Tangerine 37 45 a -lnk 5.9450) 1.77 0. .7574) 2. 1971).95 27. Q10 Species Temperature (ºC) 28 Orange 37 45 28 Lemon 37 45 28 Grapefruit 37 45 28 Tangerine 37 45 a -lnk 4. Kinetic parameters for b* values in citrus juice concentratesa.5271 (0.7997) 1.18 19.01 1.60 1.6078 (0.9657) 3.9976) 1.9896) 2.41 0.10 (0. On the other hand.9802) 2.84 (0.90 2. decomposes easily in strongly acid solution (Huelin et al.5770 (0.57 Numbers in brackets represent the determination coefficients (R2) that nonenzymatic browning reactions in orange and tangerine juice concentrates are more sensitive to temperature rises.4522 (0. the development of color in these concentrates is rapidly accelerated.Kinetics of Nonenzymatic Browning Reaction in Citrus Juice Concentrates during Storage Table 2.8551) 4.

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