Formulation and device for emitting gas to reduce physiological disorders in fruit and vegetable products

Abstract

The present invention relates to a formulation and a device containing same, which reduces physiological disorders during the transport and post-harvest storage of fruits or vegetables. The device and the formulation can release active agents in gaseous form to reduce plant tissue deterioration associated with oxidative damage at the cell membrane level. In a preferred embodiment of the invention, the active agents in the device or the formulation correspond to encapsulated compounds that allow controlled release associated with the container (device) or applied in gaseous form prior to packaging or with the packaged fruit (composition).

Description

[0001] FORMULATION AND DEVICE FOR GASEOUS EMISSION TO REDUCE PHYSIOLOGICAL DISORDERS OF HORTICULTURAL PRODUCTS

[0002] DESCRIPTIVE MEMORANDUM FIELD OF THE INVENTION

[0003] The present invention falls within the field of preserving minimally processed products in the agricultural and food industries. Specifically, the invention presents a formulation and a device for reducing the deterioration of fruits and vegetables before or during transport and post-harvest storage.

[0004] BACKGROUND

[0005] The invention relates to a formulation and a device containing it that reduces the expression of physiological disorders, including internal browning disorder (IBD), which occurs in stone fruits, such as cherries, during transport to market. IBD is a physiological disorder that manifests after prolonged storage periods, exceeding 35 days, in fruits such as cherries, plums, nectarines, and peaches, among others. The main characteristics of IBD are changes in pulp color (brownish tones), alterations in sensory characteristics, and a compromised final organoleptic quality, which is perceived by the consumer as a product with an off-flavor that is unsuitable for consumption, resulting in product loss. Figure 1 illustrates the difference between a healthy cherry and one with IBD.

[0006] Internal browning (IB) is one of the main problems facing the cherry industry in distant markets like China. It occurs in cultivars Lapins, Skeena, and Regina, which represent 36%, 2%, and 15%, respectively, of the total 415,398 tons of cherries exported by Chile in the 2022 / 23 season (Cherries Yearbook 2023, Iqonsulting). The Far East is the main destination, accounting for 92% of exports. Internal browning is considered a problem when it affects more than 15% of the total fruit in a box. The Regina variety is the most susceptible to IB.

[0007] In the case of cherries, one strategy to reduce internal browning is to use controlled atmosphere packaging, where the CO2 concentration is increased to 10% and the O2 concentration is lowered to 6%, or through modified atmosphere (MAP) packaging, where a balance of 5% CO2 and 10-12% O2 is achieved under saturation conditions. With MAP, the risk is greater because CO2 levels can rise due to a lack of temperature control during transit, leading to toxicity.

[0008] For plums, internal browning is associated with fruit harvested at a more advanced stage of ripeness. In this species, in addition to reducing the time from harvest to cooling, it is necessary to reduce the time to reach 0°C and ensure continuous storage at that temperature. Post-harvest, the gaseous product 1-methylcyclopropene (1-MCP) has proven effective in reducing the disorder in some varieties; however, in others, it causes prolonged inhibition of ripening. In peaches and nectarines, browning is the most significant disorder and develops during transport to markets. Conditioning at high temperatures (24-48 hours at 20°C) prior to cooling reduces browning but increases softening, making it commercially unviable. In other species, such as apples and pears, browning is associated with prolonged storage times and / or overripe harvesting.

[0009] STATE OF THE ART

[0010] The use of volatile compounds in post-harvest damage prevention and to prevent attacks by certain agricultural pathogens is known in the prior art. Some gaseous formulations are applied by fumigation or exposure to vapors, spraying, or immersion in a liquid containing the volatile compounds.

[0011] The following are some publications related to the field of the present invention. In the scientific publication by Neri et al. 2007 (doi.org / 10.1094 / PD-91-0030), the objective was to determine the in vitro efficacy of certain volatile plant compounds against the phytopathogenic fungus Monilinia laxa and their potential as fumigant agents for post-harvest treatment to prevent brown rot in stone fruits. In the in vitro tests, filter paper was impregnated with solutions of the compounds to be evaluated and placed on the lid of a Petri dish. In the in vivo tests, M. laxa was inoculated through a small superficial wound in the fruit, and the fruits were exposed to the vapors emitted by the liquid compounds inside a flask. Unlike the present invention, a device with an encapsulated formulation for the controlled release of a gas combination is not described.One of the advantages of the invention, with respect to this document, is the ability to obtain controlled emission profiles through a device containing a porous organic material with a high storage potential for the volatile and which, together with selective water barriers, provides a release of the volatile in low but constant concentrations during the storage time. This allows highly sensitive species or cultivars to avoid developing phytotoxicity, unwanted flavors and odors upon exposure to the volatile.

[0012] The scientific document by Neri et al. 2006 (doi: 10.1111 / j.1365-3059.2005.01312.x) aimed to determine the efficacy of certain volatile plant compounds, such as trans-2-hexenal, for controlling Penicillium expansum in vitro and to test fumigant compounds for controlling blue mold in pears. As in the previous document, for the in vitro tests, filter paper was impregnated with the compounds to be evaluated and placed on the lid of a Petri dish. In the in vivo tests, P. expansum was inoculated through a small surface wound on the fruit, and the fruit was exposed to the vapors emitted by the liquid compounds. Unlike the present invention, it does not consider compositions comprising chitosan, PVA, cyclodextrins, or cellulose, nor does it consider a gas-release matrix like the one mentioned in this inventive proposal, and it has a different technical objective.Furthermore, the concentrations evaluated are five times lower than those used for the control of internal browning and the exposure is short.

[0013] The document by Joo et al. 2012 (doi.org / 10.1016 / j.ijfoodmicro.2011.11.015) aimed to study and characterize the mechanical, physical, and antimicrobial properties of antimicrobial plastics intended for packaging. For example, the main pathogens analyzed were Alternaria solani, Aspergillus niger, Botrytis cinerea, Colletotrichum acutatum, and Penicillium sp. The effect on physiological disorders related to fruit or vegetable quality was not tested, nor was fruit browning evaluated or mentioned. Furthermore, since these are plastics, the study does not involve gaseous emission with a controlled profile of the active compounds. Using various extrusion processes significantly reduces the concentration of the active compound in the polymer matrix, making its effectiveness in controlling physiological disorders less likely. The present invention does not require these processes.

[0014] The publication by Heras-Mozos et al. 2021 (doi.org / 10.3390 / foodsl0092166) analyzed the antifungal effect of strawberry packaging systems and evaluated the fruit's external appearance. The analysis explored various methods for incorporating trans-2-hexenal into strawberry packaging, including its addition to a polymer matrix, absorption into a cellulosic medium, injection into corrugated cardboard, or as an addition to cellulosic pads. They also presented several methods for incorporating the active agent, such as encapsulation in p-cyclodextrin, inclusion in a polyamide (PA) solution, and as a coating on polypropylene (PP) film. In summary, this is a distinct technical field with a solution that does not correspond to the device according to the present invention. Meanwhile, in Chilean patent document No. 57,988, equivalent to US patent 10,449.156 B2, a controlled-release system comprising a biodegradable polycaprolactone polymer carrier and a polyhydroalkanoate polymer matrix is ​​protected. As described, the system contains encapsulated or volatile gases; however, unlike the present invention, at least one of these gases is selected from the group consisting of phenylethyl alcohol and 1-methylcyclopropene. That is, this invention differs both in the carrier material and in the active agents that constitute the composition. Regarding its use, the document does not address one of the most relevant objectives of the present invention, which is the reduction of internal browning in fruits, particularly cherries.

[0015] Although various compositions or devices for the preservation of post-harvest horticultural products are known in the state of the art, there is no composition and device or sachet with the advantages and characteristics according to the present invention.

[0016] BRIEF DESCRIPTION OF THE FIGURES

[0017] Fig. 1. Symptoms of internal browning of the pulp in cherries. Comparison of healthy fruit versus fruit with internal browning.

[0018] Fig. 2. Graphical representation of the Gas Emission Device (PAD). A: Upper cellulose sheet, B: Impregnated encapsulated active, C: Lower cellulose sheet.

[0019] Fig. 3. Characterization of the release of the encapsulated and impregnated compounds hexanal (A) and trans-2 hexenal (B) in the device expressed through the gas concentration (ppm) over time (days at 0°C) for two dosages, Dose 1 (D1:20 g) and Dose 2 (D2:40 g) in 2.5 kg modified atmosphere bags without fruit.

[0020] DETAILED DESCRIPTION OF THE INVENTION

[0021] The invention relates to a formulation and a device containing it, intended to reduce physiological disorders in horticultural products during transport and post-harvest storage. The proposed solution consists of a formulation that releases active compounds derived from the aromatic profile of the fruit. The selected active compounds reduce cell membrane deterioration and, consequently, control physiological disorders such as pulp browning. The products can be applied via a device in the packaging or as a gassing treatment prior to conventional transport of horticultural products.

[0022] Composition

[0023] The composition is made from active compounds of the C6 aldehyde type, preferably hexanal (C6HI2O, compound A) or trans-2-hexenal (C6HI0O, compound B). The composition contains chitosan, acetic acid, beta-cyclodextrin, polyvinyl alcohol, and the active C6 aldehyde compound in a 1:1 to 1:3 ratio with beta-cyclodextrin (O-cyclodextrin, O-CD) as an encapsulating agent.

[0024] The concentration ranges for each compound (%w / v) are on the order of:

[0025] • Chitosan: 0.2 - 3.0

[0026] • Acetic acid: 0.2 - 2.0

[0027] • Beta-cyclodextrin: 5.0 - 15.0

[0028] • Polyvinyl alcohol (PVA): 1.0 - 5.0

[0029] • C6 aldehyde compound: 5.0 - 15.0

[0030] The release concentration over time will depend on the amount of active ingredient and the ratio of PVA to chitosan. Increasing the concentrations of chitosan and PVA increases the release of the active compound. This behavior can be explained by the fact that chitosan helps retain certain volatile organic compounds, and in combination with an increased concentration of PVA, it reduces the water barrier, thus lengthening the emission profile.

[0031] In a preferred embodiment of the invention, the composition consists of a solution where the following compounds are combined in grams (g) in 100 ml of solution: Chitosan (0.5g), acetic acid (0.5g), Beta-cyclodextrin (12.5g), polyvinyl alcohol (PVA, 1.75g) and the active compound C6 aldehyde (5 to 15g) in a 1:1 to 1:3 ratio with the Beta-cyclodextrin.

[0032] In a preferred embodiment of the invention, the active compound C6 aldehyde is in a 1:3 ratio with Beta-cyclodextrin.

[0033] The C6 aldehyde compound is selected from either Hexanal (C6HI2O, compound A) or Trans-2-hexenal (CeHioO, compound B). The incorporation of chitosan into the composition and the confining device provides an increase in antifungal capacity, increases the degree of encapsulation of the active compound and improves the mechanical properties of the matrix, allowing for controlled release of the compound.

[0034] Device

[0035] In the case of the device, it consists of a support medium (pad) impregnated with the active compounds, which is sandwiched between two cellulose sheets that contain the active compounds within the pad and allow the passage of moisture or free water. The device is activated in the presence of free water, allowing the gaseous emission of the active compounds into the modified atmosphere packaging, where the device, through its configuration, optimizes the release of the active compound.

[0036] The active compound is pre-encapsulated before being incorporated into the support medium.

[0037] The elements that make up the device optionally correspond to organic and biodegradable components.

[0038] The device has the following main components (Fig. 2):

[0039] a. A top cellulose sheet;

[0040] b. A lower cellulose sheet;

[0041] c. An intermediate support medium containing an impregnated C6 aldehyde active compound, selected from Hexanal (C6HI2O, compound A) or Trans-2-hexenal (C6HI0O, compound B), wherein the active compound is part of a composition comprising chitosan, acetic acid, beta-cyclodextrin, polyvinyl alcohol, and the active compound in a 1:1 to 1:3 ratio with the beta-cyclodextrin,

[0042] where the gaseous emission of the compounds is activated in the presence of free water.

[0043] The intermediate support medium is a biopolymeric matrix composed, for example, of chitosan and PVA or another solid or hydrogel support material compatible with the active ingredients according to the invention. O-cyclodextrin (O-CD) is used to encapsulate the active ingredient, either hexanal (active compound A) or trans-2-hexenal (active compound B). The polymeric composition, consisting of chitosan (QN), acetic acid, and polyvinyl alcohol, has barrier properties to regulate the contact of water with the encapsulated active compound, thereby regulating the activation of the device and its gas release rate over time.

[0044] In the device, cellulose material is used as a medium to retain the impregnated compounds. Hexanal (H-AXA) or trans-2-hexenal (TH-EXA) are used as the active ingredients. The formulation (encapsulated matrix assembly) will be distributed onto the cellulose sheet by impregnation.

[0045] Advantages of the present invention

[0046] The formulation and device according to the invention, in addition to effectively controlling browning, favorably alters the organoleptic characteristics of the fruit for consumption, accentuating the sweetness and texture of the fruit.

[0047] Fruit exports are carried out using modified atmosphere packaging; this device complements that technology. Incorporating the device into traditional AM bag packaging does not alter any of the current processing steps at packing facilities.

[0048] The device is activated using free water obtained from the same line that supplies the cherry during box filling.

[0049] The device can be made with organic and biodegradable products.

[0050] EXAMPLES

[0051] Example 1: Preparation of the formulation according to the present invention

[0052] Table 2 shows the formulation according to two preferred formulas of the invention. Table 2. Formulation of the polymer composition and encapsulation of the active compounds (hexanal or trans-2 hexenal) for a 100 mL solution.

[0053] >

[0054]

[0055] Example 2. Vaporization test. Exposure of fruits to vapors in a hermetically sealed system at 0°C, prior to packaging in a controlled atmosphere (MA) bag.

[0056] Objective: To verify the effectiveness of the volatile organic compounds hexanal and trans-2 hexenal in reducing the browning of cherry pulp cv. "Regina" by exposure to vapors prior to packaging.

[0057] Methodology: Cherries cv. "Regina" were selected from a commercial harvest, from an orchard located in Molina, Maule Region, Chile. After harvest, the fruit was hydrocooled, without fungicide application, and transported to the Fruit Postharvest Physiology and Technology Laboratory, Pontifical Catholic University of Chile, where the experimental setup was carried out.

[0058] The fruit was passively cooled in a 0°C chamber. Volatile organic compounds were vaporized in airtight glass jars (4.25 L) containing 1.3 kg of fruit at concentrations of 25 and 50 ppm gas for 30 min at 0°C. At the end of the vapor exposure time, a 1 mL gas sample was extracted and analyzed using a gas chromatograph (Shimadzu GC 2014). The fruit was packed in 1.2 kg modified atmosphere (MAP) bags (Cristal Cherry, San Jorge Packaging) and stored for 45 days (d) at 0°C. A control treatment consisted of fruit not exposed to vapors and packaged directly in MAP bags. The fruit was characterized at harvest, after 45 days at 0°C (end of storage) and after 3 days at 15°C (temperature rise, simulating ambient temperature conditions). At each evaluation time (45 days at 0°C and 3 days at 15°C) the following measurements were taken:

[0059] a) Pulp browning. This parameter was evaluated on a sample of 30 fruits per replicate through a longitudinal cut and classified into internal browning (close to the pit) and sub-epidermal browning (close to the epidermis), in the categories mild (1), moderate (2), and severe (3) according to the visual severity scale. The percentage of total incidence of each type of browning, healthy fruit, and severity index was calculated. Fruits with both types of browning were considered to have mixed browning.

[0060] b) Pedicel quality. Classified according to a visual scale relative to the color of the pedicel in the categories green, yellow green, brown green and brown, value expressed as a percentage.

[0061] c) Rough skin ("Orange peel disorder"). Disorder classified according to a visual scale referring to the degree of severity of rough skin according to the affected surface of the fruit in the categories mild, moderate and severe.

[0062] d) Maturity parameters associated with fruit quality. Selection of 10 fruits per replicate. Skin color was measured with a CR-400 colorimeter (Konica Minolta Sensing Inc., Japan) and expressed as hue angle (CIEL*C*hue color space). Fruit firmness was obtained using Firmpro (HappyVolt Chile) and expressed in (gf mm 1 ). Soluble solids (%) were measured with a digital refractometer (PAL-1, Atago Co. Ltd., Tokyo, Japan) and titratable acidity (%) was determined by titration of juice using NaOH (0.1 N) until pH 8.2 was reached using a pH-211 pH meter (Hanna Instruments, Romania).

[0063] e) Analysis of Polyphenol oxidase (PPO) enzyme activity. Ten fruits were selected per replicate, frozen with liquid nitrogen, and stored in an ultra-freezer until analysis. Enzyme activity was detected with modifications according to the protocols described by Zauberman et al. (1991), Dong et al. (2015), and Kaur et al. (2019).

[0064] Results

[0065] Opening the bags after 45 days at 0°C did not reveal any fruit with symptoms of toxicity attributed to the evaluated products. Total browning was significantly reduced in the H-50 ppm and T2-H 25 and 50 ppm treatments (less than 20%) compared to the control treatment, which showed 32.5% browning after 45 days at 0°C (Table 3). Sub-epidermal browning only occurred at 15°C, also reflecting an increase in fruit with mixed browning (Table 3). The control showed 84.4% total browning, significantly higher than the Hexanal (H-25 and 50 ppm) and trans-2 hexenal (T-2H 50 ppm) treatments. The greatest reduction in total browning was observed in H-50 ppm after 3 days at 15°C (Table 3).

[0066] The severity index showed differences after 3 days at 15°C, with a high index for the control treatment in the browning and mixed browning categories, compared to the significantly lower index for the hexanal and trans-2 hexenal treatments (Table 4-5). The activity of the polyphenol oxidase (PPO) enzyme, responsible for the color change in the final stage of the disorder, was significantly higher in the control compared to the other treatments after 3 days at 15°C (Table 6), demonstrating greater cell damage in this fruit. No differences were observed in pedicel quality parameters, skin roughness, rot, or maturity parameters (color, firmness, SS, TA) that would identify a trend attributable to the application of the different treatments versus the control (Table 7-8).

[0067] Table 3. Incidence of internal, sub-epidermal, mixed, total (%) and healthy fruit (%) browning in cherries cv. "Regina" after 45 days at 0°C and 3 days at 15°C according to the different treatments of hexanal (H) and trans-2 hexenal (T-2H).

[0068] Browning (%)

[0069] Evaluation Treatment Healthy Fruit (%) Internal Sub-epidermal Mixed Total

[0070] 45 da 0°C H- 25 ppm 25.9 be 0 0 25.9 be 74.1 ab

[0071] H- 50 ppm 19.4 ab 0 0 19.4 ab 80.6 be T-2H 25 ppm 11.7 a 0 0 11.7 a 88.3 c T-2H 50 ppm 19.1 ab 0 0 19.1 ab 80.9 be Control 32.5 c 0 0 32.5 c 67.5 a p-value 0.023 0.020 0.023

[0072] 3 da 15°C H- 25 ppm 14.0 10.7 a 23.9 ab 48.6 ab 51.4 be

[0073] H- 50 ppm 16.1 7.3 a 11.5 a 35.0 a 65.0 c T-2H 25 ppm 2.2 33.2 b 36.0 b 71.4 be 28.6 ab T-2H 50 ppm 9.2 18.4 ab 23.5 ab 51.1 ab 48.9 be Control 14.6 32.4 b 37.4 b 84.4 c 15.6 a p-value 0.103 0.036 0.001 0.002 <0.001 Different letters indicate significant difference according to LSD test with p<0.05Table 4. Incidence of internal and sub-epidermal browning by severity category (%) in cherries cv. "Regina" after 45 days at 0°C and 3 days at 15°C according to the different treatments of hexanal (H) and trans-2 hexenal (T-2H).

[0074] Internal P. (%) Sub-epidermal P. (%) Evaluation Treatment

[0075] Mild Moderate Severe Total Mild Moderate Severe Total 45 da 0°C H- 25 ppm 21.7 be 4.2 0 25.9 be 0 0 0 0

[0076] H- 50 ppm 13.4 ab 4.2 1.8 19.4 ab 0 0 0 0 T-2H 25 ppm 10.0 a 1.7 0 11.7 a 0 0 0 0 T-2H 50 ppm 18.3 abe 0.8 0 19.1 ab 0 0 0 0 Control 28.4 c 3.3 0.8 32.5 c 0 0 0 0 p-value 0.029 0.201 0.542 0.023

[0077] 3 da 15°C H- 25 ppm 10.6 b 2.4 1.0 a 14.0 4.7 4.3 a 1.7 10.7 a H- 50 ppm 11.2 b 3.3 1.6 ab 16.1 4.0 2.5 a 0.8 7.3 a T-2H 25 ppm 1.5 a 0.7 0.0 to 2.2 9.9 16.5 b 6.8 33.2 b T-2H 50 ppm 7.5 ab 1.7 0.0 to 9.2 3.5 10.2 ab 4.5 18.4 ab Control 2.7 to 6.4 5.3 b 14.6 8.5 15.7 b 8.0 32.4 b p-value 0.015 0.138 0.006 0.103 0.571 0.004 0.119 0.036 Different letters indicate significant difference according to LSD test with p<0.05Table 5. Severity index (1-3) of internal, sub-epidermal and mixed browning in cherries cv. "Regina" after 45 days at 0°C and 3 days at 15°C according to the different treatments of hexanal (H) and trans-2 hexenal (T-2H).

[0078] severity index

[0079] Treatment Evaluation

[0080] Internal P. Sub-epidermal P. Mixed Pl 45 da 0°C H- 25 ppm 1.2

[0081] H- 50 ppm 1.4

[0082] T-2H 25 ppm 1.2

[0083] T-2H 50 ppm 1.1

[0084] Control 1,2

[0085] p-value 0.476

[0086] 3 da 15°C H- 25 ppm 1.3 a 1.7 ab 1.3 a H- 50 ppm 1.3 a 1.3 a 1.4 ab T-2H 25 ppm 1.3 a 1.9 b 1.6 b T-2H 50 ppm 1.1 a 2.1 b 1.5 ab Control 2.2 b 2.0 b 2.5 c p-value 0.009 0.033 <0.001 Different letters indicate significant difference according to LSD test with p<0.05

[0087] Table 6. Polyphenol oxidase (PPO) enzyme activity in cherries cv. "Regina" stored for 45 days at 0°C and then for 3 days at 15°C according to the different treatments of hexanal (H) and trans-2 hexenal (T-2H).

[0088] PPO treatment (Ul kg) 1 )

[0089] H- 25 ppm 955.0 ab

[0090] H- 50 ppm 1126.9 b

[0091] T-2H 25 ppm 897.7 a

[0092] T-2H 50 ppm 1031.4 ab

[0093] Control 1394.3 c

[0094] p-value <0.001

[0095] Different letters indicate significant difference according to LSD test with p<0.05. Table 7. Incidence of rots (%), pedicel quality according to the categories green, yellow green, brown green and brown (%) and total rough skin (%) and by categories (mild, moderate and severe) in cherries cv. "Regina" after 45 days at 0°C and 3 days at 15°C according to the different treatments of hexanal (H) and trans-2 hexenal (T-2H).

[0096] Pedicel quality (%) Rough skin (%) . Rot _ _

[0097] Green Treatment Evaluation

[0098] ' / o 'Green ... , Light Brown Moderate Brown Severe Brown Total yellow brown

[0099] 45 da 0°C H- 25 ppm 0 66.0 19.6 11.3 ab 3.1 39.6 51.0 9.4 98.1

[0100] H- 50 ppm 0 60.6 20.0 16.9 ab 2.5 51.6 39.0 9.4 92.5 T 2H 25

[0101] 2.4 60.6 16.3 20.6 b 2.5 50.5 44.9 4.6 95.0 PPm

[0102] T 2H 50

[0103] 0 70.6 18.2 9.2 to 2.0 52.1 37.9 10.0 96.3 PPm

[0104] Control 0.3 58.3 22.8 13.9 ab 5.0 49.6 43.2 7.2 96.9 p-value 0.469 0.608 0.894 0.015 0.825 0.803 0.645 0.807 0.083 3 da 15°C H- 25 ppm 2.2 to 51.0 5.6 34.5 8.9 72.8 20.6 6.6 66.2

[0105] H- 50 ppm 4.9 to 40.2 9.8 42.3 7.7 86.5 12.4 1.2 55.2 T 2H 25

[0106] 7.4 ab 37.2 10.8 41.5 10.5 75.5 16.9 7.6 61.5 PPm

[0107] T 2H 50

[0108] 17.8 b 43.3 6.0 41.7 9.0 78.0 20.3 1.7 65.7 PPm

[0109] Control 2.9 to 51.1 7.3 29.7 11.9 72.3 20.4 7.3 56.0 p-value <0.001 0.599 0.614 0.539 0.890 0.692 0.871 0.798 0.540 Different letters indicate significant difference according to LSD test with p<0.05Table 8. Characterization of the maturity parameters of Firmness, color, soluble solids (SS) and titratable acidity (TA) in cherries cv. "Regina" after 45 days at 0°C and 3 days at 15°C according to the different treatments of hexanal (H) and trans-2 hexenal (T-2H).

[0110] Evaluation Treatment Firmness (gf mm' 1 ) Color (°h) SS (%) AT (%)

[0111] 45 to 0°C H- 25 ppm 368.4 to 11.4 21.4 0.59 ab

[0112] H- 50 ppm 451.4 b 11.6 21.4 0.58 ab T-2H 25 ppm 395.8 a 12.0 20.6 0.53 a

[0113] T-2H 50 ppm 395.8 to 11.2 21.1 0.61 b Control 379.3 to 11.8 20.9 0.55 a p-value 0.002 0.621 0.728 0.006

[0114] 3 to 15°C H- 25 ppm 311.2 to 12.9 to 19.7 to 0.43

[0115] H- 50 ppm 345.1 ab 13.6 b 19.9 ab 0.43

[0116] T-2H 25 ppm 361.3 b 12.0 a 21.4 b 0.45

[0117] T-2H 50 ppm 348.9 ab 12.3 ab 20.8 ab 0.43 Control 325.1 ab 11.8 to 20.8 ab 0.47 p-value 0.041 0.006 0.181 0.815

[0118] Different letters indicate significant difference according to LSD test with p<0.05

[0119] In conclusion, the application of hexanal and trans-2-hexenal via vapor exposure prior to AM packaging significantly reduced pulp browning in CV. Regina cherries stored for 45 days at 0°C and 3 days at 15°C. Both compounds showed a significant effect on disorder control. The doses of 50 ppm hexanal and 25 ppm trans-2-hexenal were the most effective for disorder reduction, as corroborated by lower polyphenol oxidase enzyme activity. Other positive effects of the 50 ppm hexanal product included increased firmness, maintenance of harvest color, and a higher hue value.

[0120] EXAMPLES

[0121] Example 3. Gas Emission Device Test. Exposure of fruit using a gas emission device inside the packaging. Objective: To determine the effectiveness of a gas emission device of hexanal and trans-2 hexenal in cherry cv. "Regina" under simulated commercial conditions.

[0122] Methodology: The release device was fabricated in the laboratory. After several tests, an encapsulation of each of the aldehydes was successfully developed. The device consists of the active ingredient (encapsulated volatile compound) encased in a polymer composition and impregnated between two cellulose sheets (15 x 20 cm). The volatile organic compounds (hexanal and trans-2-hexenal) were used separately in the device's fabrication. The concentration of each compound was 17 mg of hexanal per gram of encapsulation and 19.4 mg of trans-2-hexenal per gram of encapsulation. The theoretical emission potential of the device for each active ingredient, for both application doses, was characterized under simulated conditions at 0°C (45 d) without fruit inside (Fig. 3). The treatments related to the gaseous emission device correspond to Hexanal and trans-2 hexanal applied in 2 doses: DI: dose 1 of 20 g D2: dose 2 of 40 g.The treatments are denoted as H-D and H-D2 (hexanal device) and T-2H D and T-2H D2 (trans-2 hexenal device), in addition to a "control" (no device). The simulation of commercial conditions consisted of selecting "Regina" cherries directly from an export line. The standard 2.5 kg AM bag box format was used. The device was placed at the bottom of the bag, replacing the usual absorbent pad, and then the box was filled with fruit. The bags were sealed and taken to the palletizing area, awaiting rapid cooling (5-7°C). The waiting time was 16 h before entering the forced-air tunnel (rapid cooling). After cooling (13 h), the fruit was held in a holding chamber and subsequently transported to the laboratory in a refrigerated truck at 0°C. oC, where 45 days of storage at 0°C was completed. Evaluations were carried out after 45 days at 0°C and 3 days at 15°C according to the criteria described for the Vaporization Test (Example 2).

[0123] Results

[0124] Opening the bags at the end of the 45 days at 0°C showed no fruit with symptoms of phytotoxicity attributed to the evaluated products. All treatments with the device significantly reduced the incidence of total browning compared to the control, which showed a high incidence at both evaluation times (Table 9). After 45 days at 0°C, the lowest incidence of total browning was obtained with the hexanal treatments (H-D1 and H-D2), with values ​​below 8%. However, after 3 days at 15°C, the greatest reduction was achieved with the trans-2 hexenal treatments (T-2H1D1 and T-2H2D2), with reductions of 58% and 60%, respectively, compared to the control (63.2%). Browning severity was higher in the control only after 3 days at 15°C in the case of mixed browning (Tables 10 and 11).The PPO enzyme activity was significantly higher in the control compared to the other treatments after 45 days at 0°C, an activity that decreased during exposure to 15°C, demonstrating the evolution of the disorder and the delay in the development of the fruit disorder with the treatments.

[0125] (Table 12).

[0126] Table 9. Incidence of internal, sub-epidermal, mixed, total (%) browning and healthy fruit in cherries cv. "Regina" after 45 days at 0°C and 3 days at 15°C according to the different treatments of hexanal (H) and trans-2 hexenal (T-2H) applied by means of the gas emission device in the container.

[0127] Internal browning (%)

[0128] Browning Treatment Evaluation

[0129]

[0130] Sub-epidermal Mixed Total

[0131] internal

[0132] 45 to 0°C H- DI 3.3 to 4.2 ab 0 7.5 to 92.5 c H- D2 4.1 to 1.7 to 0 5.8 to 94.2 c T-2H DI 5.8 ab 5.0 ab 0 10.8 ab 89.2 be T-2H D2 7.2 b 10.6 b 83.3 b Control 9.2 b 23.3 c 0 32.5 c 67.5 a p-value 0.005 <0.001 <0.001 <0.001 3 to 15°C H- Dl 5.8 24.2 b 7.5 37.5 b 62.5 b H- D27.5 , 18.8 b 30.0 ab 70.0 be T-2H DI 7.5 11.6 a 7.5 26.7 a 73.3 c T-2H D2 7.5 14.2 ab 3.3 25.0 a 74.9 c Control 8.3 44.1 c 10.8 63.7 c 36.0, p value-14 <0.001 0.203 <0.001 <0.001 Different letters indicate significant difference according to LSD test with p<0.05

[0133] No differences were observed in pedicel quality parameters, skin roughness, rot, or maturity parameters (color, firmness, SS) that could define a trend attributable to the application of the device (Tables 13 and 14). Only a greater retention of acidity was observed in all device treatments compared to the control, which showed a decrease after 3 days at 15°C (Table 14). Changes in organoleptic quality were detected in the fruit treated with the device, intensifying the characteristic cherry flavor, attributed mainly to the maintenance of fruit acidity (Table 14).

[0134] Table 10. Incidence of internal browning (%) according to different values ​​of severity of internal, sub-epidermal and mixed browning in cherries cv. "Regina" after 45 days at 0°C and 3 days at 15°C according to the different treatments of hexanal (H) and trans-2 hexenal (T-2H) applied by means of the gas emission device in the container.

[0135] Internal browning (%) Sub-epidermal browning (%) Evaluation Treatment

[0136] Mild Moderate Severe Total Mild Moderate Severe Total 45 da 0°C H- Dl 3.3 a 0 0 3.3 a 4.2 a 0.0 a 0 4.2 ab H- D2 4.1 ab 0 0 4.1 a 0.8 a 0.9 a 0 1.7 a T-2H DI 5.8 ab 0 0 5.8 ab 4.2 a 0.8 a 0 5.0 ab T-2H D2 4.2 ab 3.3 b 0 7.5 ab 5.0 a 3.3 ab 0.9 9.2 b Control 7.5 b 1.7 ab 0 9.2 b 13.3 b 9.2 b 0.8 23.3 c p-value 0.010 0.021 0.010 0.012 0.026 0.573 <0.001 3 da 15°C H- Dl 4.2 0.8 0.8 5.8 7.5 a 14.2 ab 2.5 24.2 b H- D2 1.7 2.5 0 4.2 8.3 a 9.2 a 0 17.5 ab T-2H DI 5.8 1.7 0 7.5 2.5 a 8.3 a 0.8 11.6 a T-2H D2 5.0 2.5 0 7.5 7.5 a 6.7 a 0 14.2 ab Control 1.7 4.9 1.7 8.3 17.5 b 24.1 b 2.5 44.1 c p-value 0.291 0.321 0.199 0.714 0.005 0.027 0.520 <0.001 Different letters indicate significant difference according to LSD test with p<0.05Table 11. Severity index (1-3) of internal, sub-epidermal and mixed browning in cherries cv. "Regina" after 45 days at 0°C and 3 days at 15°C according to the different treatments of hexanal (H) and trans-2 hexenal (T-2H) applied by means of the device.

[0137] severity index

[0138] Treatment Evaluation

[0139] Internal P. Sub-epidermal P. Mixed 45 da 0°C H- Dl 1.0 1.0

[0140] H- D2 1.0 1.5

[0141] T-2H DI 1.1 1.2

[0142] T-2H D2 1.4 1.6

[0143] Control 1.1 1.5

[0144] p-value 0.057 0.503

[0145] 3 da 15°C H- Dl 1.4 1.8 1.0 a H- D2 1.7 1.5 1.0 a T-2H DI 1.1 1.8 1.0 a T-2H D2 1.3 1.4 1.5 b Control 2.1 1.7 3.0 c p-value 0.103 0.117 <0.001 Different letters indicate significant difference according to LSD test with p<0.05

[0146] Table 12. Polyphenol oxidase (PPO) enzyme activity in cherries cv. "Regina" evaluated after 45 days at 0°C and 3 days at 15°C according to the different hexanal (H) and trans-2 hexenal (T-2H) treatments applied using the device.

[0147] PPO (Ul kg 2 )

[0148] Treatment

[0149] 45 days 0°C 3 days 15°C

[0150] H- Dl 1356.1 be 1146.0 d

[0151] H- D2 1241.5 b 1069.6 cd

[0152] T-2H DI 764.0 to 916.8 be

[0153] T-2H D2 592.1 to 840.4 ab

[0154] Control 1528.0 c 706.7 a

[0155] p-value <0.001 <0.001

[0156] Different letters indicate significant difference according to LSD test with p<0.05. Table 13. Incidence of rots (%), pedicel quality according to the categories green, yellow green, brown green and brown (%) and total rough skin (%) and by categories (mild, moderate and severe) in cherries cv. "Regina" after 45 days at 0°C and 3 days at 15°C according to the different treatments of hexanal (H) and trans-2 hexenal (T-2H) applied by means of the device.

[0157] Pedicel quality (%) Rough skin (%)

[0158] " . . , T Rot

[0159] Green Treatment Evaluation

[0160] ' 1Green ... . Forgive Mild Moderate Severe Total

[0161] gray yellow

[0162] co n

[0163] 45 to 0°C H- DI 0.4 to 59.8 to 5.5 ab 31.4 c 3.3 be ¿ 17.3 0.0 80.3 ab

[0164] ab

[0165] H- D2 0.4 a 86.5 c 3.2 a 9.8 ab 0.5 ab 65.8 b 17.5 0.0 83.3 ab

[0166] T-2H DI 1.3 b 79.8 be 7.5 b 12.7 ab 0.0 a 63.5 b 18.3 0.0 81.8 ab

[0167] T-2H D2 0.7 a 70.8 b 7.2 b 16.7 b 5.3 c 54.5 a 18.5 1.3 74.3 a

[0168] son

[0169] Control 0.4 to 84.8 c 8.5 b 6.5 to 0.2 ab 24.8 0.8 85.9 b

[0170] ab

[0171] p-value 0.001 <0.001 0.010 <0.001 0.007 0.010 0.418 0.259 0.043

[0172] 3 to 15°C H- Dl 0.5 25.4 2.7 ab 55.0 16.9 be 63.3 14.0 to 0.8 to 78.1

[0173] H- D2 0.2 29.7 5.1 b 60.0 5.2 to 60.6 22.3 ab 0.9 to 83.8

[0174] T-2H DI 0.2 35.4 1.7 a 53.3 9.5 ab 52.6 26.5 b 0.5 a 79.6

[0175] T-2H D2 0.7 26.5 1.8 to 49.7 22.0 c 61.8 23.5 ab 1.2 to 86.5

[0176] Control 0.3 33.1 2.1 ab 61.0 3.8 a 62.6 18.9 ab 4.3 b 85.8

[0177] p-value 0.581 0.577 0.049 0.24 0.004 0.639 0.280 <0.001 0.470

[0178] Different letters indicate significant difference according to LSD test with p<0.05. Table 14. Characterization of the maturity parameters of Firmness, color, soluble solids (SS) and titratable acidity (TA) in cherries cv. "Regina" after 45 days at 0°C and 3 days at 15°C according to the different treatments of hexanal (H) and trans-2 hexenal (T-2H) applied by means of the device.

[0179] Firmness

[0180] Evaluation Treatment Color (°h) SS (%) AT (%) (gf mm 1 )

[0181] 45 days at O°C H- DI 471.3 10.8 20.5 0.82 H- D2 496.3 10.5 20.6 0.83 T-2H DI 480.1 11.2 20.6 0.82 T-2H D2 480.2 20.6 0.82 Control 493.7 11.2 20.5 0.80 p-value 0.779 0.448 0.999 0.788 3 days at 15°C H- DI 465.0 11.7 ab 22.1 c 0.69 c H- D2 472.0 12.0 b 20.1 ab 26.0 ab T-H DI 492.1 11.5 to 21.5 be 0.62 ab T-2H D2 501.2 11.5 to 20.9 be 0.64 be Control 502.9 11.9 ab 19.6 to 0.57 a p-value 0.295 0.026 <0.009 Letters indicate significant differences significant according to LSD test with p<0.05

[0182] In conclusion, the application of hexanal or trans-2 hexenal compounds by incorporating the device inside the packaging in AM significantly reduced internal browning of 'Regina' cherries. No differences were found between the hexanal (H) and trans-2 hexenal (T-2H) compounds, nor between the doses of the active ingredient evaluated. Another positive effect observed with both types of devices was the preservation of the characteristic cherry flavor, attributed primarily to the maintenance of acidity during high-temperature ripening.

Claims

CLAIMS 1. A composition for reducing physiological disorders in horticultural products during transport and post-harvest storage, CHARACTERIZED in that it comprises a C6 aldehyde compound, in addition to chitosan, acetic acid, beta-cyclodextrin, polyvinyl alcohol and wherein the C6 aldehyde compound is in a 1:3 ratio with the beta-cyclodextrin.

2. A composition according to claim 1, CHARACTERIZED in that the concentration ranges of each compound correspond to: • 0.2 - 3.0 %w / v Chitosan, • 0.2 - 2.0 %w / v Acetic acid, • 5.0 - 15.0 %w / v Beta-cyclodextrin, • 1.0 - 5.0 %w / v Polyvinyl alcohol (PVA), and • 5.0 - 15.0 %v / v active compound C6 aldehyde, where the active compound C6 aldehyde is in a 1:1 to 1:3 ratio with beta-cyclodextrin.

3. A composition according to claim 1 or 2, CHARACTERIZED in that the C6 aldehyde compound is selected from Hexanal (C S HI2O, compound A) or Trans-2-hexenal (CgHioO, compound B).

4. A composition according to any of claims 1 to 3, CHARACTERIZED in that the active C6 aldehyde compound is in a concentration of 5 to 15% w / v.

5. A gaseous emission device for reducing physiological disorders in horticultural products during transport and post-harvest storage, CHARACTERIZED in that it comprises: a. A top cellulose sheet; b. A lower cellulose sheet; c. An intermediate support medium containing a composition comprising a C6 aldehyde compound, chitosan, acetic acid, beta-cyclodextrin, polyvinyl alcohol and the active compound in a 1:1 to 1:3 ratio with beta-cyclodextrin, wherein the gaseous emission of the compounds is activated in the presence of free water.

6. The device according to claim 5, CHARACTERIZED in that the intermediate support medium is composed of a solid or hydrogel type biopolymeric matrix.

7. The device according to claim 5, CHARACTERIZED in that the intermediate support medium according to (c) is composed of one or more sheets of a biopolymeric matrix consisting of chitosan and PVA.

8. The device according to claim 5, CHARACTERIZED in that the C6 aldehyde compound is selected from Hexanal (C S HI2O, compound A) or Trans-2-hexenal (CgHioO, compound B) 9. Use of a composition as defined in any of claims 1 to 4, CHARACTERIZED in that the composition is used to reduce internal browning in fruit during transport and post-harvest.

10. Use of a device as defined in any of claims 5 to 8, CHARACTERIZED in that the device is used to reduce internal browning in fruit during transport and post-harvest.

11. Use according to claim 10, CHARACTERIZED in that the device is used to reduce internal browning in cherries.

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