POSTHARVEST MODIFICATIONS OF MECHANICALLY INJURED BANANAS

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Saavedra del Aguila, Juan; Sichmann Heiffig-del Aguila, Lília; Fumi Sasaki, Fabiana;

Mitsuyuki Tsumanuma, Guy; Graças Ongarelli, Maria das; Fillet Spoto, Marta Helena;

Jacomino, Angelo Pedro; Marcos Ortega, Edwin Moisés; Kluge, Ricardo Alfredo

POSTHARVEST MODIFICATIONS OF MECHANICALLY INJURED BANANAS.

Revista Iberoamericana de Tecnología Postcosecha, vol. 10, núm. 2, 2010, pp. 73-85

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Postharvest modifications of Mechanically… Juan Saavedra del Aguila y cols. (2010)

POSTHARVEST MODIFICATIONS OF MECHANICALLY INJURED BANANAS Juan Saavedra del Aguila1,6; Lília Sichmann Heiffig‐del Aguila2; Fabiana Fumi Sasaki1,8; Guy Mitsuyuki Tsumanuma3; Maria das Graças Ongarelli1; Marta Helena Fillet Spoto4; Angelo Pedro Jacomino3,7; Edwin Moisés Marcos Ortega5; Ricardo Alfredo Kluge1,7 1Departament of Biological Science, Escola Superior de Agricultura “Luiz de Queiroz” (ESALQ), University of São Paulo (USP), C.P. 9, 13418‐900, Piracicaba, SP, Brazil. E‐mail: [email protected]; 2Grains and Fibers Center, Agronomic Institute (IAC), C.P. 28, 13075‐630, Campinas, SP, Brazil; 3Departament of Crop Production, ESALQ – USP; 4Departament of Agro industry, Food and Nutrition, ESALQ – USP; 5Departament of Basic Science, ESALQ – USP; 6FAPESP fellow; 7CNPq fellow; 8CAPES fellow. Key words: Musa acuminata AAA cv. Nanicão, ripening, color, damage, postharvest

ABSTRACT The major problem affecting bananas (Musa spp.) during and after harvest is the susceptibility of the mature fruit to physical damage caused during transport and marketing. The purpose of this study was to evaluate the effects of mechanical injuries on physiological, physicochemical and anatomical parameters of banana fruits. The treatments were: non‐injured fruit (control), three 60 cm free falls, three longitudinal cuts (70 mm long and 2 mm deep), three longitudinal scratches on the edges (50 mm long and 2 mm wide), and compression for 15 minutes (equivalent force of 52.9 Newton). Fruits were stored for 21 days at 25oC and 75% RH. The parameters analyzed were the respiratory rate and ethylene production, loss of fresh mass, pulp/skin relation, total soluble solids (TSS), total acidity (TA), TSS/TA ratio, total amount of carotenoids, ascorbic acid amount, firmness, color (L*, chroma and hue) and evaluated by electronic scanning microscopy. Fruits from all treatments showed a respiratory peak on the nineteenth day and a decrease in this variable afterwards. The kind of injure may anticipate by one (impact injures) or two (cut and scratching injuries) days the ethylene production peak, which occurs on the 15th day after harvest. It was observed that the cutting and scratching treatments were responsible for the most undesirable changes in fruits, such as increased loss of fresh mass and changes in color (darkening), when compared to the control and the other treatments. Consequently, cutting and scratching injuries can be considered the most harmful postharvest mechanical injuries for bananas.

MODIFICACIONES POST‐COSECHA DE BANANA DAÑADA MECÁNICAMENTE Palabras clave: Musa acuminata AAA cv. Nanicão, maduración, color, daño, post‐cosecha

RESUMEN El mayor problema que afecta los frutos de banana (Musa spp.) durante y después de la cosecha, es la susceptibilidad de los frutos maduros a los daños físicos causados durante el transporte y la comercialización. El objetivo de este estudio fué evaluar los efectos de los daños mecánicos sobre las variables fisiológicas, físico‐químicas y anatómicas en frutos de banana. Los tratamientos fueron: fruto sin daño mecánico (control), tres caídas libres desde 60 cm, tres cortes longitudinales (70 mm largo y 2mm ancho), tres raspados longitudinales (50mm largo y 2mm ancho), y 15 minutos de compresión (con una fuerza equivalente de 52,9 Newton). Los frutos fueron almacenados por 21 días a 25oC y 75% HR. Las variables respuesta analizadas fueron la taza respiratoria, producción de etileno, pérdida de masa fresca, relación pulpa/casca, sólidos solubles totales (SST), acidez total (AT), “ratio” SST/AT, carotenoides totales, ácido ascórbico, firmeza, color (L*, “chroma” y “hue”) y evaluaciones con microscopio electrónico de barredura. Los frutos de todos los tratamientos presentaron su pico respiratorio en el día 19 de experimento y con decrecimos de taza respiratoria a partir de este día. El pico de etileno fue adelantado en un día (daño por impacto) o en dos días (daños por corte y raspado), en los otros tratamientos este pico ocurrió en el día 15 de experimento. Fue observado que los tratamientos de corte y de raspado fueron los que presentaron los cambios más indeseables en los frutos, como el incremento de la pérdida de masa fresca y modificaciones en el color (pardeamiento), cuando los anteriores tratamientos fueron comparados al control y a los demás tratamientos. Consequentemente, las injurias por corte y raspado pueden ser consideradas las más perjudiciales injurias mecánicas en la post‐cosecha de frutos de banana.

Rev. Iber. Tecnología Postcosecha Vol 10(2):73-85 73


INTRODUCTION Banana (Musa spp.) is one of the most consumed fruit worldwide and is grown in most tropical countries. In Brazil, nearly all bananas produced are consumed fresh, playing an important role in feeding low‐income families, providing a source of income to them and keeping the rural labor force in the country. Banana is a major fruit regarding people’s nutrition, not only due to its highly nutritious value, but also due to its low production cost. According to data from 2004, Brazil is nowadays the second world producer of bananas, while India is by far the largest producer (FAO, 2005). The growing Brazilian production of bananas mainly results from successive changes in cultivation techniques taking place nowadays. However, the development of conservation techniques and quality control have not improved at the same rhythm. Only 50 to 60% of the bananas produced in Brazil reach the end consumers due to high losses during the commercialization process (Mascarenhas, 1999). Banana is a climacteric fruit that presents high respiratory rate and ethylene production after harvesting, which makes it highly perishable (Pinheiro et al., 2005). Mechanical injuries may be defined as plastic deformations, superficial ruptures and destruction of vegetal tissues led by external factors. Such injuries lead to physical modifications (physical damages) and/or physiological, chemical and biochemical alterations that alter color, aroma, flavor and texture of vegetables (Mohsenin, 1986). Injuries can be classified as compression, impact or cut. Impact is generally caused by the collision of the fruit against solid surfaces or against other fruits during harvest, handling and transport. Compression injuries are caused by a variable pressure on the fruit surface exerted by an adjacent fruit or by the container holding the fruits. Cut injuries are

generally due to the collision of the fruit against a sharp surface that ruptures the epidermis, or due to the pressure exerted by uneven surfaces, such as the container edges, against the fruit (Mattiuz and Durigan, 2001; Chitarra and Chitarra, 2005). Mechanical injuries are among the main factors affecting postharvest losses in bananas. Different injuries may cause different effects on agricultural products, mainly changes in color and appearance, fast ripening (due to increased respiratory rate and ethylene production), increase in loss of water and in deterioration by microorganisms, thus, directly affecting fruit quality and retail prices (Dadzie and Orchad, 1997; Lladó and Dominguez, 1998). Mishandling, vibration, impacts, compression and/or superficial bruises are the basic causes of banana fruit injuries leading to fruit deterioration and favoring the development of diseases (Cortez et al., 2002). Chitarra (1998), describing the effect of cuts and injuries on plant cell membranes, comments that such injuries lead to the rupture of organelles, modify cell permeability and favor cell disorganization, triggering the ethylene synthesis and increasing respiration rates. Upon cutting, the free (polyunsaturated) fatty acids from banana react with the O2 through lipoxygenases forming hydroperoxides that lead to losses in the nutritional value, to detrimental alterations in taste and aroma and to the formation of dark pigments. The induction of the ethylene synthesis and the increase in respiration rates cause variations in the maturation rates of injured and intact tissues, increasing water loss by exudation, accelerating defense reactions in tissues, deteriorating fruit quality and reducing their shelf life. The purpose of this study was to evaluate the alterations in banana fruits, cv. Nanicão (Musa acuminata, AAA), Cavendish group, submitted to four different kinds of



mechanical injuries and stored at room temperature. MATERIAL AND METHODS Plant material and Treatments Harvesting of banana fruits cv. Nanicão was carried out in banana orchards in Piracicaba (SP) region when the central fruit of the second bunch showed 34±2 mm diameter. Fruits were immediately and carefully taken to the Laboratory. Fruits were, then, selected for firmness (touch), absence of mechanical injuries and visible infections. Later, only the second, third and fourth bunches were selected, from which fruits were removed and individualized. A completely randomized experimental design was used, with 6 replicates for treatments analyzing carbon dioxide (CO2) and ethylene (C2H4) and 4 replicates for the other evaluations was used. The samples of CO2 and C2H4 answers variables were the same ones since the beginning at end experiment, for the amount these analyses were no destructive and the samples were not independent among; on the other hand, the other answers variables, the analyses were destructive in each evaluation period (time) and for the amount existing independence among the samples in the time. The treatments were chosen in function of the importance in the banana postharvest; in the case of the Free fall treatment, for example, doing a pre‐test of application of this treatment, with the objective that the treatment is not discarded the fruit treated immediately, the Free fall of the fruit to larger heights of 60 cm, caused the instance fruit discard for the immediate rupture of the peel fruit. Treatments were: T1 = non‐injured fruits (control), T2 = 60 cm free fall (three impacts per fruit at the same place on the median region, exactly in the middle of the concave side), T3 = three longitudinal cuts (70 mm long and 2 mm deep) on the median region,

T4 = three longitudinal scratches (50 mm long and 2 mm wide) on median edges, in the corners of the fruit, and, T5 = mechanical compression on the median region for 15 minutes (equivalent force of 52.9 Newton (N)). Injured areas were demarked and fruits were placed on expanded polystyrene trays or glass flasks, according to their destination and stored at room temperature (25oC (±2oC) and 75% (±5%) RH). Such environment conditions were daily monitored using a Temptec thermohydrographer. Assessments In order to evaluate the respiratory rate, banana fruits (weighing around 150 g) were placed in hermetic glass flasks (1693.5 mL) and stored at room temperature, 25oC (±2oC) and 75% (±5%) RH. A silicon septum was inserted into the lid of each flask and an aliquot (1 mL) of the internal atmosphere was taken through it. Gas samples taken from each container through the silicon septum were injected into a gas chromatographer Trace 2000 GC (Thermoffinigan) equipped with a 2m‐Porapack N column and a flame ionization detector (FID). Hydrogen was used as the carrier gas at 40 mL min‐1. Temperatures used in the equipment were 100oC in the column, 100oC in the injector, 250oC in the detector and 350oC in the CO2 methanator. Calibration standards for assessing carbon dioxide (CO2) were 2150 μL L‐1 and 29900 μL L‐1 CO2. The results in % CO2 were used to calculate the respiratory rate, considering the flask volume, the fruit mass and the period of time flasks remained closed. A gas sample was taken from flasks one hour after conducting the treatments and the evolution of CO2 was evaluated, corresponding to time‐zero (0) reading. Posterior readings were conducted daily for 20 days for the five treatments. Results were expressed in mL CO2

kg‐1 h‐1.



Ethylene production was quantified by using the same samples and the same procedures regarding sampling and reading. Temperatures in the column, injector and detector were 100oC, 100oC and 250oC, respectively. The calibration standard to measure C2H4 amounts was 1.94 μL L‐1 C2H4. Results were expressed in mL C2H4 kg

‐1 h‐1. Run time for both gases (CO2 and C2H4) was 1 minute. The loss of fresh mass was determined by the difference between the final mass and the initial mass of each replicate and results were expressed in % of fresh mass loss. The pulp/skin mass relation was also determined. The amount of total soluble solids (TSS) was determined after triturating each sample in a household multiprocessor. Then, a drop of the triturated mass was placed in an Atago manual digital refractometer and results were expressed in ºBrix (Carvalho et al., 1990). The tritratable acidity (TA) was determined by titration with NaOH 0.1 N to pH 8.1, and results were expressed as [H+] in (mol L‐1) (Carvalho et al., 1990). The ratio (TSS/TA) was also calculated. The total skin carotenoids amount was determined by spectrophotometry. Five grams of the sample triturated in a mortar with 3 g of Hyflosupercel (celite) and 50 mL of cold acetone was filtered (vacuum) using a Buchner funnel with filter paper. Part of the filtered sample was placed in a separation funnel containing 40 mL of petroleum ether and 300 mL of distilled water was poured on the funnel walls to avoid the development of an emulsion and to maintain two separate phases (petroleum ether + carotenoids, and water + acetone). The aqueous phase was then discarded and another portion of the filtered sample was added, repeating the same procedure with 200 mL of distilled water (the process was repeated three times). After complete acetone removing, the petroleum ether phase was filtered through a glass filter containing 15 g of anhydrous sodium sulfate to

remove the water, placed in a volumetric balloon (50 mL) and completed with petroleum ether to 50 ml volume. Readings were conducted in a spectrophotometer (FEMTO‐700 Plus), absorbance range of 450 nm, using the petroleum ether as blank. Readings were used to determine the total carotenoids amount using the formula:

Total Carotenoids = [A*volume (mL)*104]/[A1%1cm*sample

weight (g)]

Where A=absorbance; volume=sample volume (50 mL); A1% 1cm =coefficient of absorption of β‐carotene in petroleum ether (2592). Results were expressed in μg g‐1 (Rodriguez‐Amaya, 2001). Ten grams of sample material was put in an erlenmeyer flask containing 50 mL of oxalic acid solution to determine the ascorbic acid amount. Titration was carried out using 2,6‐Dichlorophenolindophenol (DCPIP) sodium salt as an indicator until a persistent pinkish color was achieved for 15 seconds (Carvalho et al., 1990). Results were expressed in mg of ascorbic acid per 100 g of sample. Pulp firmness was evaluated near the injured or not (about 2 cm) using the manual penetrometers Fruit Pressure Tester FT 011 and FT 327 (diameter = 80 mm) and results were expressed in Newtons (N). Color parameters, such as luminosity (L*), a* and b* values, were determined using a colorimeter (Minolta CR‐300), near the injured or not (about 2 cm). Results were used to calculate hue angle (color) and the corresponding saturation (Chroma), as recommended by Minolta (1994). Reading were performed on the banana skin, comprising an average of four readings per fruit. The median region and the surroundings of mechanically injured areas were identified for the conduction of readings. Evaluations of the carbon dioxide (CO2) and ethylene (C2H4) productions were carried out daily for 20 days, while evaluations or other



parameters were carried out every three days for 21 days. In order to better visualize the injures, tissue samples from the median region and the surroundings of mechanically injured areas were fixed in 2.5% glutaraldehyde buffer solution of sodium cacodylate 0.05 mol L‐1, pH 7.2, for 72 hours at 5ºC and analyzed in an electronic scanning microscope. Next, samples were washed five times in sodium cacodylate buffer solution 0.05 mol L‐1, pH 7.2, and fixed in osmium tetroxide 1% for one hour. Fruits were then dehydrated in a gradual series of acetone solutions (30%, 50%, 70% 90% and 100%). Samples were dried to critical point using liquid CO2 and metallized with gold to enable the analysis under electronic scanning microscope LEO 435VP – ZEISS. Data analysis The experimental design adopted was the completely randomized with 5 treatments and 6 (CO2 and C2H4) our 4 (others parameters) replicates for each treatment. Results of weigth loss, pulp/skin, TSS, TA, ratio, carotenoids total, ascorbic acid, firmness, L*, Chroma and hue angle were submitted to analysis of variance by F test and comparison of means by Tukey test at 5% probability. Regression modules were applied only for the respiratory rate and ethylene production of the fruits, selecting the models REG, GLM and MIXED, using the statistic software SAS. GLM proc was used to verify the sphericity supposition and F tests valid for the intra‐individuals factors with the due corrections for the number of freedom degrees. The model used to model the repeated data measures in the time, it is the mixed model that it is specified for: 1,2,...m,j , =++= jjjjj bZXY εβ

In that Yj dimension (nj x1) it is a vector of the m individuals' answers along the time, Xj dimension (nj x p) it is the planning head office

corresponding to the fixed effect, β dimension (p x 1) it is the vector of the coefficients of the regression of the population average, call of fixed effects, Zj dimension (nj x q), it is the planning head office corresponding to the vector of random effects bj , dimension (q x 1) and εj dimension (nj x 1) of random mistakes. It was used proc MIXED to esteem the parameters of the model. For the storage time for being a quantitative factor was used a model of regression polynomial and a test of comparison of averages for treatment factor (Tukey‐Kramer). RESULTS AND DISCUSSION Fruits from all treatments showed a respiratory peak on the nineteenth day and a decrease in this variable afterwards (Figure 1). Mattiuz and Durigan (2001), studying mechanical injuries in guavas stored at 23.4oC, observed an increase in the respiratory activity throughout the storage period, regardless of injuries and cultivars used, though not pinpointing the respiratory peak of the fruits analyzed. Control fruits showed the lowest respiratory rates during the evaluation period, reaching 12.14 mL CO2 kg

‐1 h‐1 on the 6th day of evaluation. On the other hand, fruits submitted to impact showed increasing rates in the respiratory activity one hour after the conduction of the treatment, reaching 74.4 mL CO2 kg‐1 h‐1. This treatment showed the highest respiratory rate throughout the experiment, reaching 144.68 mL CO2 kg

‐1 h‐1 on the 19th day of evaluation (Figure 1), the fruits didn’t present visible rots in this treatment and in the rest of treatments. The free fall treatment altered the respiratory metabolism of fruits. On the first and second days of evaluation, this treatment showed respiratory rates statistically superior to the other treatments, which leveled with the cut treatment afterward. The respiratory rate for the free fall treatment on the 20th day



was superior only to the compression treatment. The respiratory rates for fruits submitted to cuts and scratching were higher than those for the control along virtually all the experiment period. Figure 1 – Polynomial regression representation of respiration rate of evolved CO2 (mL CO2 kg

‐1 h‐1) in banana fruits under mechanical injuries, during storage at 25oC (±2oC) and 75% (±5%) RH. The respiratory intensity is a key factor determining the shelf life of vegetables after harvest, once respiration oxidizes reserve substances, leading to the senescence of organs (Wills et al., 1981). The ethylene production did not show statistical differences up to the 13th day of evaluation, when there was an ethylene peak for the treatments involving cutting and scratching, which reached 49.54 and 45.29 mL C2H4 kg

‐1 h‐1, respectively. The same was observed for the free fall treatment on the 14th day, when the ethylene production was 42.10 mL C2H4 kg

‐1 h‐1. Ethylene production values for the control and compression treatment were 24.73 and 50.16 mL C2H4 kg

‐1 h‐1, respectively, with production peaks on the 15th day of evaluation. The highest ethylene production peak was verified for the compression treatment (Figure 2). Formulas of polynomial regression for each treatment of the variables respiratory rate and ethylene production(Y) in day relation (X) are showed in the table 1. Images from the electronic scanning microscopy evidenced the anatomical

differences in fruits undergoing the different treatments tested. Such differences influenced the physiological and physical behavior of banana fruits. Figure 3A (control) shows totally turgid epidermal cells of the banana skin, differently from what can be observed in Figure 3B (free fall treatment), where cells are wilted and display membrane injuries. Figure 3C shows intact epidermal cells in the non‐injured part of fruits from the cutting treatment, as well as totally dilacerated epidermal parenchyma cells from injured areas of fruits from the same treatment. Figure 3D (scratching treatment) shows totally injured epidermal cells (signed by arrows) and an open stomatal pore probably due to the loss of the ability to control the opening and closing of injured guard cells, which leads to high percentages of fresh mass loss in the scratching treatment. The compression treatment (Figure 3E) caused a certain turgidity loss in epidermal cells. Figure 2 ‐ Polynomial regression representation of ethylene production rate (mL C2H4 kg

‐1 h‐1) in banana fruits under mechanical injuries, during storage at 25oC (±2oC) and 75% (±5%) RH. Highly significant differences in fresh mass loss were observed from the 12th day of evaluation when comparing the cutting and scratching treatments with the other treatments. Such pattern persisted until the end of the experiment. The fruits fresh mass losses from treatments involving cutting, scratching, control, compression and free fall

Scratches

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

-2 -1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21

Free

mmLL

CCOO

22 kkgg-- 11

hh--

Cuts

Control

Compression

days

-5

5

15

25

35

45

-2 -1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21

Cuts Free fall

Compression Control Scratchets

days

mmLL

CC22HH

44 kkgg-- 11

hh-- 11




were 44.8%, 41.9%, 25.9%, 24.6% and 22.1%, respectively (Table 2). In fact, such differences were expected for this variable, once the treatments carried out damaged the epidermal skin cells, affecting their function and exposing larger areas of skin to water loss. Theses injuries were more severe in the

cutting and scratching treatments due to the rupture of cells and, specifically in the scratching treatment, to the damage to stomatoes, which made them lose the ability to control the transpiration process, remaining open, as observed in the microphotographs (Figure 3).

Table 1 – Formulas of polynomial regression for each treatment of the variables respiratory rate and ethylene production(Y) in day relation (X).

Treatment Yn = β0 + β1 Xn+ β2 Xn

2 + β3Xn3 R2

Y = Respirtory rate (mL CO2 kg‐1 h‐1)

Control Yco = 38.538 + (‐7.786) Xco + 0.515 Xco2 + (‐0.00002) Xco

3 0.824Free fall Yf = 54.020 + (‐11.927) Xf + 0.882 Xf

2 + 0.00005 Xf3 0.732

Cuts Ycu = 41.185 + (‐9.054) Xcu + 0.771 Xcu2 + (‐0.09) Xcu

3 0.789Scratchets Ys = 36.388 + (‐6.835) Xs + 0.440 Xs

2 + 0.02 Xs3 0.836

Compression Yc = 37.214 + (‐7.838) Xc + 0.549 Xc2 + (‐0.002) Xc

3 0.812 Y’ = Ethyelene production (mL C2H4 kg

‐1 h‐1)Control Y’co = 4.489 + (‐3.882) X’co + 0.617 X’co

2 + (‐0.021) X’co3 0.470

Free fall Y’f = 4.367 + (‐4.110) X’f + 0.681 X’f2 + 0.022 X’f

3 0.409Cuts Y’cu = 6.270 + (‐6.020) X’cu + 0.978 X’cu

2 + (‐0.033) X’cu3 0.517

Scratchets Y’s = 5.428 + (‐5.230) X’s + 0.865 X’s2 + (‐0.030) X’s

3 0.505Compression Y’c = 4.656 + (‐3.629) X’c + 0.530 X’c

2 + (‐0.016) X’c3 0.487

A B

C D

E

A B

C D

E

Figure 3 – Electronic scanning microscope pictures of the treatments: (A) control; (B) free fall; (C) cuts; (D) scratches and (E) compression of the peel in banana fruits under mechanical injuries, during storage at 25oC (±2oC) e 75% (±5%) RH. Bar = 20 μm. (day 21).


Maia et al. (2004), studying metabolic alterations due to mechanical damages in bananas known as dwarf Prata, observed that injuries anticipated the climacteric peak and the ripening of fruits. Considering the percentage of fresh mass loss, damages

caused by cutting and scratching led to a higher percentage loss, while those caused by compression showed a similar index to the control fruits.

Table 2 ‐ Weigth loss (%), pulp/skin mass relation and total soluble solids (TSS) in banana fruits under mechanical injuries, during storage at 25ºC (±2ºC) and 75% (±5%) RH. Standard error in parenthesis.

Treatment Days of storage

0 3 6 9 12 15 18 21 Weigth loss

Control 0.0 a E* (0.00)

4.7 c DE (0.17)

7.9 b CDE (0.32)

12.2 bc BCD (1.21)

13.6 b BC (0.77)

18.9 c AB (1.10)

19.9 c AB (1.21)

25.9 b A (4.23)

Free fall 0.0 a E (0.00)

5.0 c DE (0.34)

10.3 b CDE (1.00)

12.6 bc BCD (0.76)

16.7 b ABC (1.05)

22.8 bc AB (4.05)

26.5 bc A (4.79)

22.1 b AB (1.01)

Cuts 0.0 a F (0.00)

9.6 a E (0.17)

15.3 a DE (0.47)

20.2 a D (0.58)

28.5 a C (1.51)

30.7 ab C (0.59)

38.5 ab B (2.66)

44.8 a A (1.91)

Scratchets 0.0 a C (0.00)

10.3 a CB (0.59)

15.7 a B (0.73)

17.7 ab B (2.50)

30.9 a A (3.21)

37.4 a A (1.74)

42.9 a A (5.80)

41.9 a A (1.86)

Compression 0.0 a F (0.00)

6.8 b E (0.14)

9.0 b DE (0.29)

11.1 c D (0.47)

16.1 b C (1.33)

17.5 c C (0.86)

21.2 c B (0.81)

24.6 b A (0.38)

Pulp/skin

Control 1.3 a B (0.06)

1.3 a B (0.04)

1.4 b B (0.02)

1.6 b AB (0.11)

1.9 a AB (0.22)

2.5 a A (0.44)

2.0 a AB (0.15)

2.5 b A (0.19)


1.3 a E (0.06)

1.7 ab DE (0.07)

1.8 ab CD (0.10)

1.9 aABCD (0.07)

2.4 a A (0.12)

2.3 a AB (0.17)

2.2 c BC (0.04)

Cuts 1.3 a E (0.06)

1.3 a E (0.06)

1.6 b DE (0.05)

2.0 ab CD (0.08)

2.1 a BC (0.07)

2.5 a B (0.09)

2.5 a B (0.13)

3.3 a A (0.17)

Scratchets 1.3 a D (0.06)

1.6 a CD (0.08)

2.0 a BCD (0.17)

2.2 a BC (0.14)

2.1 a BCD (0.10)

2.6 a AB (0.11)

2.6 a AB (0.34)

3.1 ab A (0.24)

Compression 1.3 a C (0.06)

1.6 a BC (0.13)

1.4 b C (0.04)

1.8 ab BC (0.13)

2.0 a BC (0.21)

2.4 a AB (0.42)

2.1 a BC (0.05)

3.1 ab A (0.12)

TSS (ºBrix)

Control 3.2 a B (0.20)

5.2 a B (0.99)

5.1 a B (0.73)

5.0 b B (0.77)

10.1 ab AB (2.78)

18.5 ab A (2.84)

18.8 c A (1.66)

21.8 ab A (1.06)

Free fall 3.2 a B (0.20)

3.7 a B (0.36)

5.0 a B (0.23)

4.2 b B (0.17)

6.9 ab B (1.50)

11.1 b AB (2.73)

20.9 bc A (2.01)

13.2 b AB (0.71)

Cuts 3.2 a C (0.20)

3.8 a C (0.38)

6.2 a C (0.55)

4.4 b C (0.39)

5.6 b C (0.43)

17.3 b B (3.83)

28.5 a A (0.40)

28.3 a A (0.67)


4.4 a C (0.48)

10.6 a BC (3.24)

20.5 a AB (1.95)

20.9 a AB (3.78)

27.7 a A (0.48)

26.9 ab A (0.68)

24.2 a A (2.10)

Compression 3.2 a B (0.20)

4.3 a B (0.18)

5.4 a B (0.79)

4.2 b B (0.31)

4.7 b B (0.83)

11.0 b B (1.23)

10.8 d B (1.09)

25.0 a A (0.44)

* Averages followed by at least one common letter, same small letter in the column and capital letter in the line, for each treatment or evaluation time, do not differ among each other, by the Tukey test (p ≤ 0.05).

The pulp skin‐1 mass relation varied from 1.3 at the beginning of the experiment to 3.3, 3.1, 3.1, 2.5 and 2.2 on the 21st day of treatments involving cutting, compression, scratching, control and free fall, respectively. On the same day, the free fall treatment showed relations significantly lower than

those for control, cutting, compression and scratching treatments (Table 2). The pulp skin‐1 mass relation seems to be determined by the water content of fruits. The increased pulp skin‐1 mass relation derives from variations in sugar concentrations in fruit tissues, which increase quicker in the pulp, leading to a



differential in osmotic pressure. As a consequence, the water present in the fruit skin is transferred to the pulp, altering this relation, which ranges from 1.2 to 1.6 in unripe fruits and from 2.0 to 2.7 in ripe fruits (Loesecke, 1950). The TSS amount in bananas increases quickly as fruits ripen due to the degradation of starch into soluble sugars. The TSS amount showed a highly significant increase between

the first and the 21st day in all treatments, except for the free fall treatment, in which this increase was lower (Table 2). The titratable acidity increased throughout the experiment. For all treatments, 2.0 mol L‐1 was initially observed, while on the 21st day, values were 5.0, 4.7, 4.4, 4.0 and 3.3 mol L‐1 for the impact, compression, control, cutting and scratching treatments, respectively (Table 3).

Table 3 ‐ Tritratable acidity as [H+] (mol L‐1), ratio (TSS/TA) and carotenoids total in banana fruits under mechanical injuries, during storage at 25ºC (±2ºC) and 75% (±5%) RH. Standard error in parenthesis.


0 3 6 9 12 15 18 21 [H+] (mol L‐1)

Control 2.0 a B* (0.16)

2.2 b B (0.12)

2.5 ab B (0.20)

2.5 b B (0.06)

3.3 a B (0.70)

5.8 b A (0.96)

6.3 a A (0.54)

4.4 ab A (0.36)

Free fall 2.0 a C (0.16)

2.1 b C (0.14)

2.8 ab BC (0.15)

2.7 b BC (0.11)

3.6 a ABC (0.50)

4.4 b ABC (0.58)

5.3 ab A (0.57)

5.0 a A (0.59)

Cuts 2.0 a B (0.16)

2.0 b B (0.06)

2.7 ab B (0.15)

2.7 b B (0.12)

3.0 a B (0.20)

6.5 a A (1.40)

3.8 b B (0.19)

4.0 ab B (0.10)


2.7 a BC (0.14)

4.3 a ABC (0.98)

5.8 a A (0.76)

4.3 a ABC (0.60)

4.2 b ABC (0.03)

3.5 b ABC (0.10)

3.3 b BC (0.05)


2.4 b B (0.03)

2.1 b B (0.20)

2.6 b B (0.20)

2.7 a B (0.36)

3.4 b B (0.31)

4.2 b A (0.35)

4.7 ab A (0.40)

(TSS/TA)

Control 16.4 a B (2.00)

22.7 a B (3.25)

20.4 ab B (1.1,9)

19.9 b B (2.77)

26.0 b B (3.54)

30.2 b AB (2.70)

30.3 b AB (3.01)

49.9 b A (2.56)

Free fall 16.4 a AB (2.00)

17.3 a AB (1.04)

17.9 b AB (0.55)

15.7 b B (0.98)

18.4 b AB (1.56)

24.0 b AB (1.20)

42.3 b A (3.74)

25.1 c AB (1.33)

Cuts 16.4 a B (2.00)

18.9 a B (1.82)

23.3 ab B (1.61)

16.1 b B (1.28)

18.2 b B (0.44)

27.2 b B (2.55)

76.1 a A (3.05)

71.5 a A (2.23)


16.0 a D (1.66)

23.2 ab CD (1.98)

38.3 a BC (3.53)

45.4 a BC (2.87)

66.5 a AB (1.49)

76.2 a A (2.57)

72.6 a A (3.25)


18.0 a B (0.81)

25.8 a B (2.09)

15.9 b B (0.44)

17.1 b B (1.05)

29.5 b B (1.1,4)

26.4 b B (1.94)

54.7 ab A (4.39)

µg total carotenoids/g

Control 13.5 a E (0.09)

12.2 c F (0.07)

13.4 d E (0.06)

17.1 b C (0.10)

19.4 c A (0.08)

18.1 c B (0.02)

18.1 b B (0.07)

15.6 c D (0.05)


13.5 a E (0.01)

15.9 c D (0.08)

17.0 b C (0.08)

19.3 c B (0.04)

19.7 b A (0.06)

16.2 c D (0.13)

16.1 b D (0.01)

Cuts 13.5 a E (0.09)

13.0 b F (0.06)

11.5 e G (0.08)

7.7 d H (0.08)

24.8 a B (0.05)

26.0 a A (0.04)

15.4 d D (0.07)

19.3 a C (0.03)


13.1 b E (0.02)

19.2 b B (0.02)

15.4 c C (0.03)

20.5 b A (0.07)

15.5 e C (0.12)

12.8 e E (0.06)

10.5 e F (0.02)

Compression 13.5 a E (0.09)

11.5 d F (0.03)

20.0 a A (0.16)

18.0 a C (0.20)

18.6 d B (0.03)

17.8 d C (0.09)

18.7 a B (0.10)

14.5 d D (0.02)


Unlike other fruits, banana fruits present low acidity at the beginning of the maturation stage and figures increase slowly as ripening

progress. When fruits are ripe, decreasing figures are observed.



Ratio figures increased throughout the experiment, ranging from 16.4 on the first day of experiment to 72.6, 71.5, 54.7, 49.9 and 25.1 on the 21st day for the scratching, cutting, compression, control and impact treatments, respectively, when the impact treatment was significantly inferior when compared to the other treatments (Table 3). A major reduction in the total carotenoid amounts was observed in the cutting treatment from the first day (13.5 μg g‐1) to the 9th day of storage (7.7 μg g‐1). After that, figures generally increased until the end of the storage stage, reaching 19.3 μg g‐1 on the 21st day of storage (Table 3). The reduction in carotenoid amounts for this treatment during the nine first storage days may have occurred due to the severity of the cutting injuries promoting greater exposure of fruit tissues to light and oxygen, which are factors responsible for carotenoid degradation (Klein et al., 1985). Moreover, the cutting injuries used favored greater loss of cell juice and, probably, greater loss of carotenoids stored in chromoplasts. On the other hand, increased amounts of carotenoids after the 9th day of storage may have been due to carotenoid synthesis in the fruit skin. According to Rodriguez‐Amaya (2001), the carotenoids synthesis may happen after harvest, although opposite results were found by Loesecke (1950), according to whom the yellow banana pigments (carotenoids) remain rather constant throughout fruit ripening. This discrepancy may well be due to different methodologies used for the carotenoids quantification. Differences for carotenoid amounts were verified among the remaining treatments. However, an increase in carotenoid amounts from 13.45 μg g‐1 on the first day of storage to 19.3, 15.6, 16.1, 10.5 and 14.5 μg g‐1 on the 21st day was verified for the cutting, control, free fall, scratching and compression treatments, respectively. The ascorbic acid amount remained stable throughout the storage period and no

differences were observed among treatments. On the 21st day, values were 3.8, 2.2, 1.3, 1.2 and 1.1 mg 100g‐1 for the cutting, scratching, compression and free fall treatments, respectively (Table 4). The amounts of ascorbic acidy found in our study were similar to those found by Aldemaro (1981), who performed a physicochemical characterization of some cultivars of bananas in Venezuela. Multiple functions are attributable to L‐ascorbic acid in humans. It is impostant for its buffer function in oxidation reduction processes, but also because of its molecular structure particularities in its ability for ions and hydrogen electrons transfer in reversible processes (Souza et al., 2008). Pulp firmness decreased by 54.0 N, on the first day, to 28.9, 10.7, 6.4, 4.5 and 2.8N, on the 21st day, for the free fall, control, compression, cutting and scratching treatments, respectively (Table 4). Bleinroth (1985) stated that the banana pulp is composed of a great number of small cells. In unripe fruits, each of these cells presents a rigid membrane, mainly composed by insoluble substances (protopectin) and numerous solid grains of starch within the membrane. During the ripening process, the protopectin is partially transformed by the action of enzymes, forming soluble pectin, which is responsible for the softening of membrane cells. At the same time, the starch is enzymatically transformed in soluble sugars, which begin to spread throughout the solid matter within the cells, yielding a semisolid mass. As a consequence of the chemical alterations that soften the cell membrane, partially dissolving its content, the extremely firm and unripe fruit turns into a soft and tasty ripe banana. The cutting and scratching treatments showed a decrease in luminosity (L*) values during storage, while the same values increased when the control and the other treatments were considered. The lowest figures were observed for the cutting and




scratching treatments on the 21st day, 32.6 and 34.1, respectively, being significantly lower than those for the control, free fall and compression treatments which were 52.4, 50.0 and 51.3, respectively. The lower luminosity values for cutting and scratching

treatments mean that the fruits skin darkened, probably due to enzymatic action, as a consequence of the injuries to which fruits were submitted (Table 5).

Table 4 ‐ Ascorbic acid (mg ascorbic acid/100g) and firmness (Newtons) in banana fruits under mechanical injuries, during storage at 25ºC (±2ºC) and 75% (±5%) RH. Standard error in parenthesis.


0 3 6 9 12 15 18 21 Ascorbic acid (mg/100g)

Control 1.7 a A* (0.11)

1.6 a A (0.12)

1.3 a A (0.01)

1.6 b A (0.13)

1.5 a A (0.12)

1.4 a A (0.09)

1.9 a A (0.12)

1.3 b A (0.18)

Free fall 1.7 a AB (0.11)

1.5 a AB (0.18)

1.8 a AB (0.19)

2.0 ab A (0.21)

1.4 a AB (0.01)

1.4 a AB (0.05)

1.6 a AB (0.10)

1.1 b B (0.03)

Cuts 1.7 a B (0.11)

1.9 a B (0.10)

2.1 a B (0.21)

2.4 a AB (0.24)

1.8 a B (0.18)

1.5 a B (0.12)

1.5 a B (0.10)

3.8 a A (0.17)

Scratchets 1.7 a A (0.11)

1.8 a A (0.08)

1.9 a A (0.19)

2.2 ab A (0.16)

1.3 a A (0.11)

2.1 a A (0.11)

1.3 a A (0.07)

2.2 b A (0.11)

Compression 1.7 a ABC (0.11)

1.3 a BC (0.01)

1.6 a ABC (0.11)

2.0 ab AB (0.01)

2.0 a A (0.14)

1.3 a BC (0.14)

1.3 a BC (0.17)

1.2 b C (0.11)

Firmness (Newtons)

Control 56.4 a A (3.33)

56.4 a A (5.33)

56.8 a A (1.11)

60.0 a A (1.73)

53.3 a AB (1.76)

22.6 a ABC (1.40)

15.6 ab C (1.88)

10.7 ab C (1.68)

Free fall 56.4 a A (3.33)

62.9 a A (6.83)

56.6 a A (2.73)

59.5 a A (5.38)

53.0 a A (5.18)

49.2 a AB (1.59)

11.5 b B (1.48)

28.9 a AB (1.45)

Cuts 56.4 a AB (3.33)

59.1 a A (3.26)

58.5 a A (1.71)

60.8 a A (1.89)

70.5 a A (6.81)

29.2 a BC (1.26)

4.1 b C (0.23)

4.5 b C (0.05)


54.3 a A (4.78)

37.5 a AB (1.65)

8.0 b BC (2.48)

14.5 b BC (1.17)

2.5 b C (0.08)

2.0 b C (0.05)

2.8 b C (0.13)

Compression 56.4 a A (3.33)

44.8 a A (4.31)

54.8 a A (5.5)

59.8 a A (4.39)

61.5 a A (2.60)

45.6 a A (3.43)

38.7 a A (1.80)

6.4 bB (0.47)


The initial Chroma (C) value of 29.0 on the first day changed to 31.3, 30.5, 27.3, 10.1 and 8.6 for the control, compression, free fall, scratching and cutting treatments, respectively, on the 21st day of storage (Table 5). Highly significant differences for hue (h) values between the cutting and scratching treatments, as well as among the other treatments, were observed on the 21st day of storage (Table 5). Within the experimental conditions, the respiratory rate of fruits increased from the 13th day of experiment, regardless of the

injuries applied, reaching a respiratory peak on the 19th day after harvest. The kind of injure may anticipate by one (impact injures) or two (cut and scratching injuries) days the ethylene production peak, which occurs on the 15th day after harvest. Banana fruits showed an increase in the total carotenoids amounts during the ripening process, whether fruits were mechanically injured or not. According to our study in function of the results of the ethylene production, weight loss, firmess, L*, Chroma and hue, the cutting and the scratching treatments caused the most undesirable alterations in banana fruits, when compared


to the control and the other treatments (free fall and compression). Consequently, these kinds of injuries should be strongly avoided during postharvest of banana fruits.

ACKNOWLEDGEMENTS We would like to thank Dr. E.W. Kitajima (NAP‐MEPA) for the use of microscope electronic; Dr. Cristina Vieira de Almeida and Dr. Beatriz Appezzato da Glória for helpful comments in the writing of this paper.

Table 5 ‐ Luminosity (L*), chroma and hue angle in banana fruits under mechanical injuries, during storage at 25ºC (±2ºC) and 75% (±5%) RH. Standard error in parenthesis.


0 3 6 9 12 15 18 21 L*

Control 49.9 a A* (0.84)

49.3 a A (0.47)

50.2 a A (0.69)

50.6 a A (0.63)

50.8 a A (2.53)

49.4 ab A (1.04)

51.2 a A (1.44)

52.4 a A (1.85)


49.4 a AB (0.31)

50.0 a A (0.67)

50.0 a A (0.55)

49.9 a A (0.50)

50.1 a A (1.11)

42.0 b B (3.88)

50.0 a A (1.53)

Cuts 49.9 a A (0.84)

49.7 a A (0.07)

49.0 a A (0.57)

48.3 a A (0.36)

47.1 a AB (0.45)

47.5 ab AB (0.57)

39.9 b BC (3.09)

32.6 b C (2.54)

Scratchets 49.9 a AB (0.84)

49.6 a AB (0.56)

52.8 a A (2.06)

53.4 a A (1.50)

52.3 a A (1.83)

42.8 b BC (2.83)

35.8 b CD (4.80)

34.1 b D (4.43)


52.5 a A (0.94)

50.6 a A (0.77)

51.0 a A (0.74)

49.6 a A (0.31)

50.6 a A (1.08)

50.7 a A (1.37)

51.3 a A (0.93)

Chroma

Control 29.0 a A (0.48)

28.9 a A (0.33)

28.8 a A (0.53)

29.1 a A (0.52)

31.0 a A (1.85)

28.5 a A (1.29)

30.5 a A (1.15)

31.3 a A (1.84)


28.2 a A (0.24)

28.3 a A (0.16)

29.4 a A (0.29)

29.2 a A (0.81)

29.1 a A (0.92)

19.3 b B (1.97)

27.3 a AB (1.75)

Cuts 29.0 a A (0.48)

28.9 a A (0.30)

27.3 a A (0.38)

28.4 a A (0.34)

26.9 a A (1.01)

27.6 ab A (1.47)

18.0 b B (1.07)

8.6 b C (1.03)


29.6 a A (0.55)

31.3 a A (1.40)

31.6 a A (1.00)

31.6 a A (2.00)

20.3 b B (3.81)

11.9 b BC (1.56)

10.2 b C (1.07)


30.1 a A (0.51)

29.3 a A (0.39)

29.4 a A (0.39)

29.0 a A (0.77)

28.6 a A (0.73)

29.1 a A (1.20)

30.5 a A (0.81)

Hue angle

Control 116.3 a A (0.42)

118.0 a A (0.12)

116.4 a A (0.51)

112.8 a A (1.17)

108.5 a A (5.03)

102.2 b A (3.47)

99.3 c A (0.90)

95.8 b A (0.91)

Free fall 116.3 a A (0.42)

118.0 a A (0.27)

114.0 a A (0.65)

112.8 a A (0.38)

108.1 a A (0.93)

145.1 b A (4.83)

170.0 b A (4.53)

96.3 b A (1.39)

Cuts 116.3 a B (0.42)

116.7 a B (0.37)

113.1 a B (0.14)

108.0 a B (0.30)

103.0 a B (0.67)

96.7 b B (1.5)

255.9 a A (6.00)

231.6 a A (3.35)

Scratchets 116.3 a B (0.42)

117.6 a B (0.24)

108.3 a B (2.24)

102.4 a B (2.27)

142.6 a B (4.91)

261.9 a A (3.73)

236.5 a A (4.26)

222.1 a A (4.91)


115.3 a A (0.94)

115.2 a A (0.48)

111.7 a A (1.91)

107.5 a A (1.91)

145.4 b A (3.33)

140.0 bc A (4.19)

179.6 a A (5.39)


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Documents

POSTHARVEST MODIFICATIONS OF MECHANICALLY INJURED BANANAS