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Article

Using the Essential Oils of Sage and Anise to Enhance the Shelf Life of the Williams (sin. Bartlett) Pear

by
Mira Elena Ionica
1,
Andrei Bita
2,* and
Felicia Tutulescu
1
1
Faculty of Horticulture, University of Craiova, 13 A. I. Cuza Street, 200585 Craiova, Romania
2
Faculty of Pharmacy, University of Medicine and Pharmacy of Craiova, 2 Petru Rareş Street, 200349 Craiova, Romania
*
Author to whom correspondence should be addressed.
Coatings 2024, 14(1), 10; https://doi.org/10.3390/coatings14010010
Submission received: 25 November 2023 / Revised: 16 December 2023 / Accepted: 19 December 2023 / Published: 20 December 2023
(This article belongs to the Special Issue Functional Coatings in Postharvest Fruit and Vegetables)
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Abstract

:
The effects of post-harvest spray treatments with essential oils (EOs) obtained from sage and aniseed on maintaining the quality of pears of the ‘Williams’ variety during storage was studied. Harvested pears were picked when they had reached their optimal maturity and underwent a treatment involving the application of aqueous solutions of glycerin, with varying amounts of sage essential oil (SEO) and aniseed essential oil (AEO). Weight loss during storage varied according to the treatment applied with the lowest values recorded for sage essential oil at concentrations of 300 ppm (6.24%) and 250 ppm (6.60%), respectively. Aniseed essential oil had a smaller effect on weight loss compared to sage essential oil. Fruit firmness was better maintained under the influence of the essential oil treatments, with those treated with sage essential oil standing out. The concentration of the essential oils that is used influences the antimicrobial activity of the post-harvest treatment that is applied, with higher essential oil concentrations leading to more pronounced decreases in the total number of mesophilic aerobic bacteria immediately after treatment (4.05 for SEO 200; 3.00 for SEO 300, respectively). The use of post-harvest techniques involving the application of aqueous solutions containing glycerol and essential plant oils by spraying can extend the shelf life of pear fruits.

1. Introduction

Due to the increasing resistance of microorganisms to various chemicals as well as the current trend to consume healthier food products without chemical residues, re-searchers are looking for new sources of antimicrobial products [1]. These can be used as replacements for various chemical treatments applied during the storage of fruit and vegetables to enhance their shelf life.
Essential oils (EOs) are complex natural compounds obtained from plants. They are characterized by their volatility and strong odor as secondary metabolites of aromatic plants. The history of essential oils began in the Orient; because the process of obtaining them by steam or hydro-distillation was first devised and used in Egypt, Persia and India [2]. Essential oils (EOs) are extracted from leaves, stems, flowers, seeds, fruit peels, etc., and are used for their olfactory, aromatic and antimicrobial properties in products such as cosmetics, foods and medicines [3].
The precise way in which essential oils function is not entirely comprehended, but some researchers have asserted that essential oils derived from plants possess the capability to disrupt the cell walls and cytoplasmic membranes of fungi and bacteria [4]. EOs can also inhibit the synthesis of proteins and polysaccharides as well as DNA and RNA in fungal and bacterial cells [5].
Salvia officinalis L. is a widely used herb in the food industry containing high levels of EOs. The antibacterial activity of essential sage oil (SEO) is widely known, the effect being based on the presence of: α-thujone, α-pinene, camphor, 1-octen-3-ol and 1,8-cineole [6,7,8]. SEO exhibits strong antimicrobial activity, especially for meat products, against: Listeria monocytogenes, Bacillus cereus, Bacillus megatherium, Bacillus subtilis, Aeromonas hydrophila, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus sp. and Klebsiella oxytoca [6,9,10,11].
SEO, extracted from the leaves of the Salvia officinalis plant, has gained attention for its diverse applications in recent years. Rich in bioactive compounds such as camphor and 1,8-cineole, sage oil exhibits notable antimicrobial and anti-inflammatory properties [12]. Modern research has highlighted its potential cognitive benefits, suggesting a positive impact on mental clarity and concentration [13]. Moreover, studies have explored the oil’s effectiveness in skincare, attributing its antibacterial properties to potential applications in addressing skin issues [14]. The versatile nature of sage essential oil finds expression in aromatherapy, where its herbaceous and sweet fragrance enhances focus and relaxation. Its contemporary relevance extends to topical applications and formulations in personal care products, showcasing the evolving understanding of its therapeutic potential. SEO also has a high antioxidant activity and can be used as a natural preservative in food products [15].
Pimpinella anisum L., (anise) is an aromatic herb containing about 6% essential oils (at least 20 mL/kg), depending on the material and extraction [16]. Anise essential oil (AEO) contains mainly trans-anethole, followed by estragole, limonene and pinene [17]. The essential oils of some anise species are used as sedatives, antidepressants and carminatives [18]. AEO has antimicrobial, antifungal and antioxidant properties [19,20,21,22]. Lately, AEO has been reported as a replacement for antibiotics in broiler feed rations due to its content of anethole, eugenol, methylchavicol, anisaldehyde and estragole [23]. Some researchers have considered that AEO, through its antimicrobial activity, can be used as a preservative for the preservation of plant and animal products [24]. Other authors showed that anise extracts had an effect against Streptococcus mutans (S. mutans) and Lactobacillus rhamnosus species [10]. Furthermore, anise essential oil has found a place in modern aromatherapy, where its characteristic licorice-like aroma is harnessed for its soothing and calming effects. This versatile oil is commonly utilized in diffusers and massage oils, reflecting a contemporary understanding of its therapeutic benefits and its integration into holistic wellness practices. AEO can also be used as a natural means of fighting insect vectors [25].
The pear (Pyrus communis L.) is a species native to Eastern Europe and Asia Minor and has been known since 3000 B.C. and was cultivated and spread by the Romans. Research on the mitosis and meiosis of pear species has indicated that these species are diploid 2n = 34 and some cultivars are triploid [26].
The pear is a tree adapted to temperate climates and the fruit has a delicate, pleasant taste, a fine texture and a subtle aroma [27]. Although it’s the fifth most cultivated fruit in the world, a large quantity of pears is lost annually due to storage (8%–29% loss) [28]. The quality of pears is a combination of organoleptic, chemical and nutritional properties as well as shelf life. The long-term cold storage of pears presents many risks leading to the loss of fruit quality, hence the need to find solutions to practical problems during such periods.
Many studies have been carried out on improving the shelf life of pears during storage using various methods [29,30,31,32] and new storage technologies have also been developed [33]. However, there are still several aspects in terms of quality deterioration of pears during storage caused by microorganisms, manipulation and physiological derangements.
In this study we aimed, for the first time, to study the effect of post-harvest treatment by spraying with EOs obtained from sage and aniseed on maintaining pear quality during storage. We also investigated the effects of the applied treatments on the storability of pears.
In our study, our objective was not to produce controlled release emulsions or gels with the utilized essential oils. Instead, the primary aim of our research was to describe a simple and practical method that could be easily employed under normal use and storage conditions. We intentionally focused on developing an approach that aligns with everyday applications, aiming for user-friendly and accessible methods. By steering away from complex formulations like controlled release systems, our intention was to provide a straightforward and versatile technique that can be readily adopted in various contexts without the need for specialized conditions or equipment. This emphasis on simplicity and applicability underscores the practical utility of our proposed method in real-world scenarios.

2. Materials and Methods

2.1. Chemicals

Reagents used: Solvents (n-hexane, water—LiChrosolv® grade) were all purchased from Merck (Burlington, MA, USA). Sodium carbonate and Methanol have been acquired from Merck Darmstadt, Germany; Folin–Ciocalteu reagent, Gallic acid, Sodium acetate, 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and 6-hydroxy2, 5, 7, 8-tetramethylchroman-2-carboxylic acid (Trolox) have been acquired from Sigma-Aldrich (Taufkirchen, Germany). The other chemicals were of analytical grade.

2.2. Plant Material

Pears belonging to the ‘Williams’ (sin. Bartlett) variety from the Pomiculture Research and Development Station Vâlcea, Romania (45°14′09.3′′ N, 24°36′67.5′′ E) were harvested at maturity and brought to the laboratory of the Faculty of Horticulture, University of Craiova, Romania. The ‘Williams’ variety is an early maturing cultivar, the fruit reaching consumption maturity between 15 August and 5 September with a storage period of about 2 months. The fruit originates from trees grafted with the intermediary, respectively: Williams/Curé/Quince A, cultivated intensively, with a distance of 3.5 m between rows and 2.5 m between the trees per row. The branches of the trees were formed in a tiered palm system with oblique arms. The age of the plantation is 10 years and during cultivation, all technological cultivation steps were carried out, namely: fruit cutting, fruit thinning, irrigation (30 L water/tree/wet), phase-fertilization (fertigation). Basic fertilization was carried out with NPK complex fertilizers (5 tons/hectare) and during vegetation, disease control treatments were carried out with Dagonis fungicide (BASF, Bucharest, Romania) in doses of 1.2 L/ha.
Dried sage (Salvia officinalis L.) and anise (Pimpinella anisum L.) plants grown in the same area (100 g for each sample), mixed with 1000 mL of water were used for the extraction of essential oils by hydrodistillation. The EO extraction was done in a Clevenger apparatus at a temperature of 102 °C for 180 min. The volume of oil obtained was 1.1–1.3 mL/100 g dry matter. The obtained EO was kept at a temperature of 4 °C until use.

2.3. Experimental Design

Pears at harvest maturity were sorted (uniform color and size) and treated by spraying with aqueous glycerol solutions (2.5%) in which various amounts of SEO and AEO were incorporated. After treatment, the pears were kept in a draught at room temperature for shriveling, packed in 14 kg capacity boxes and stored at a temperature of 3 ± 1 °C and 80% relative humidity. 56 kg of pears (4 wooden boxes) were used for each of the different treatments.
Depending on the type of EO and the concentrations used, several experimental variants were designed (Table 1).
The concentrations of EOs used were based on previous research [34]. Several analyses and determinations were performed: weight loss, firmness, dry matter (DM), total soluble content (TSS %), total phenols content (TP mg GAE/100 g fw), antioxidant activity (AOA %), skin color during storage.
The action of EOs on the microbiological load was shown by the variation of the total number of aerobic mesophilic bacteria (TAMB log CFU/g) on the fruit surface during storage. EOs obtained by hydrodistillation were also analyzed to identify the main components. Also, at the end of the storage period a sensory analysis of the fruit was performed.

2.4. Analytical Methods

The EOs were obtained using an assembly comprising a heating mantle (NAHITA, Barcelona, Spain) directly connected to a Neo Clevenger type continuous distiller (Glasco, Seaside, OR, USA), as previously described [35]. The extracted oils were injected undiluted into the GC-MS.
The GC/MS analysis aimed to identify the constituents present in the anise and sage EOs through a comparative elucidation of its mass spectrum with a reference spectrum (NIST Library version 2020). The analysis was performed on a Thermo Scientific Focus GC equipped with an AI/AS 3000 autosampler, coupled with a DSQ II mass detector, and featuring a TraceGOLD TG-624 column (60 m × 0.25 mm × 1.4 μm). The injection volume, at a flow rate of 1.3 mL/min with a split ratio of 1:50, was 1 μL, employing helium as the carrier gas.
The initial oven temperature was set at 110 °C and maintained for 5 min, followed by an increase to 220 °C at a rate of 2 °C/min, sustained for 15 min. The MS transfer line temperature was held at 240 °C, the ion source temperature at 230 °C, and the electron impact ionization (EI) was configured at 70 eV. Spectra were examined in the full scan mode covering the range of 50 to 450 mass, and the retention times (RI) of all constituents were recorded.
Weight loss was assessed by weighing the pears with a digital scale (Sartorius 157 CP124S, Epsom, UK, accuracy = 0.01 g) both at the beginning of the experiment and every 30 days of storage and the results obtained were expressed as percentage weight loss from initial weight [34].
Dry matter (DM %) is a recently established quality indicator because it is a function of starch that hydrolyzes over time into soluble sugars during the storage and ripening process. Measured at harvest and during storage, it indicates internal quality after prolonged periods of cold storage. DM was assessed through gravimetric evaluation by subjecting 5 g of finely divided fresh pears to drying in a laboratory oven (Memmert, Germany) set at 105 °C until a consistent weight was achieved. The outcomes were then presented as a percentage of dry matter [36]. For determining the total soluble solids content (TSS %), a digital refractometer (Hanna Instruments, Woonsocket, RI, USA) was employed. A sample obtained by blending the pears in an electric mixer was used, and the results were presented as a percentage of total soluble solids [34]. To characterize the internal quality of the pears, the firmness of the fruits was determined from the opposite parts of each fruit, with a digital texturometer for vegetables and fruits (Multilab, Baia de Aramă, Romania) provided with a penetrometric ponson of 8 mm in diameter.

2.5. Total Phenolic Content Evaluation

The total phenolic content was determined using the previously described Folin–Ciocalteu method [37]. In this procedure, three grams of pear homogenate underwent extraction with 10 mL of methanol for 60 min using an ultrasonic bath at room temperature, followed by centrifugation at 6000 RPM for 15 min. The resulting supernatants were collected and stored at −40 °C. For the assay, 100 μL of the extract was combined with 5 mL of distilled water, and 500 μL of Folin–Ciocalteu reagent was added, followed by the addition of 1.5 mL of sodium carbonate solution (20% w/v). The mixture was then brought to a total volume of 10 mL with distilled water, vigorously shaken, and incubated in the dark at 40 °C for 30 min.
Using a Varian Cary 50 178 UV spectrophotometer (Varian Co., Palo Alto, CA, USA), the absorbance was measured at 765 nm. The results were quantified in milligrams of Gallic acid equivalent (GAE) per 100 fresh weight (f.w.), determined based on a calibration curve previously constructed using standard Gallic acid solutions.

2.6. Antioxidant Activity Evaluation

The assessment of antioxidant activity (AOA) was conducted following the procedure outlined by Oliveira et al., with certain modifications, specifically focusing on the free radical scavenging activity of extracts against the DPPH free radical [37]. For this evaluation, 50 μL of pear extract, obtained as described for the determination of total phenol content, was combined with 3 mL of a 0.004% DPPH methanolic solution. The mixture was vigorously shaken and left in the dark for 30 min. Subsequently, the absorbance was measured at 517 nm using a Varian Cary 50 UV-VIS spectrophotometer. To establish a baseline, a blank sample was created by blending methanol with the DPPH solution. The results (%) were calculated using the following formula:
DPPH scavenging activity (%) = [1 − absorbance of sample/absorbance of blank] × 100.

2.7. Color Evaluation

Fruit skin color was measured on the most colored part of fruits at the midpoint between the stem and calyx. The evaluation of pear color involved the measurement of parameters such as brightness (L*), redness (a*), yellowness (b*), chroma, and hue angle within the CIELab system. This analysis was conducted using a Thermo Scientific Evolution 600 UV/VIS spectrophotometer, employing 1 mm quartz cuvettes.
From these measurements, both the yellowness index and the yellow color value were calculated following specific formulas [38]:
Yellowness index = 142.86 b*/L*; Yellow color value = (a*/b*).

2.8. Bacteriological Analysis

The determination of aerobic mesophilic bacteria was conducted following the protocol outlined by Aycicek, Oguz, and Karci [39]. Ten grams of pears was blended with 90 mL of a dispersal solution (0.1% peptone water) for 120 s. An inoculum of 0.1 mL from dilutions of 1/1000 and 1/10,000 was applied to the surface of the standard culture medium (Granucult Merck, Burlington, MA, USA) spread on regular plates. Temperature control was maintained at 30 °C for 2 days. The quantitative assessment was carried out through colony counting. The results, expressed as log CFU/g, were derived from the mean of three replicates and underwent statistical validation. Conventional tests involving morphology, dye characteristics, and cultural methods were employed for identification purposes [40].

2.9. Sensory Analysis

Sensory evaluation of pears was carried out at the end of the storage period with the help of volunteer students from the Faculty of Horticulture of the University of Craiova. Each volunteer individually completed an organoleptic tasting sheet. The evaluation was performed using a 10-point hedonic scale, 1 meaning “I don’t like it at all” and 10 meaning “I like it completely”. Aspects considered were taste, color and overall acceptability. Volunteers were asked to rinse their mouths with distilled water before each sample tasted. Volunteers who carried out the tasting received no information about the samples they evaluated, which were coded with letters and numbers. As the institution does not have an ethics committee for sensory evaluation of products, all participants signed a written consent.

2.10. Statistical Analysis

The obtained results underwent statistical analysis using Statgraphics Centurion Version XVIII software (Statgraphics Technologies, Inc., The Plains, VA, USA). All tests and analyses were conducted in triplicate, and the mean values for each sample were employed in the statistical evaluation. To assess significant differences between the means of the utilized treatments and the three storage periods, a one-way ANOVA with a significance level of p < 0.05 was applied to test the null hypothesis. Subsequently, a least significant difference (LSD) test was performed to identify statistical differences at the 0.05 level of significance.
For a comprehensive overview of dataset variability, standard deviation was calculated using Microsoft 365 Excel, and the data are presented as means ± SD.

3. Results

3.1. Components of Sage and Anise Essential Oils

The sage and anise essential oils were analyzed in order to identify the main components (Figure 1 and Figure 2). Further analysis of gas chromatography data can provide valuable insights into the terpene composition of these essential oils. By comparing the results of the gas chromatography analysis, we can assess the similarities and differences in the volatile compounds present in these two essential oils (Table 2). Gas chromatography analysis of anise essential oil has shown that the major component present is anethole, which accounts for a significant percentage of the oil composition. On the other hand, gas chromatography analysis of sage essential oil revealed a different composition, with its own unique set of volatile compounds. The sage oil contains thujone, a compound known for its potential medicinal properties and distinct aroma. Other sage oil compounds are also present in smaller quantities, including 1,8-cineole (eucalyptol), camphor and α-pinene [41,42,43,44,45].

3.2. Weight Loss

The weight loss of pear fruit increased directly proportionally to the storage time (Figure 3). At the end of storage, weight loss was higher in the control (7.98%) and glycerol-treated (7.77%) variants than in the variants treated by spraying with aqueous solutions of essential oils and glycerol. The results obtained are consistent with those presented by Adhikary et al. (2022) [46] and with the normal shelf life of Williams pears (4–8 weeks) [47,48]. In terms of differences between the treatments applied, the lowest weight loss was observed with SEO 300 (6.24%) followed by SEO 250 (6.60%), SEO 200 (7.30%) and AEO 300 (7.31%).

3.3. Firmness

At the beginning of the experiment the fruit firmness was 4.385 kgf/cm2. It did not change immediately after treatment with essential oils and the dryness of the fruit. The results of fruit firmness indicate that ripening was significantly (p < 0.05) faster in the control and G 2.5 samples than in the samples treated with SEO and AEO (Figure 4). Pear firmness decreased with storage time, the results obtained being in accordance with those found by Goke, Serra and Musacchi (2020) [49].
Among the treatments, control (1.529 kg/cm2) and G 2.5 (1.733 kg/cm2) pears exhibited, after 3 months of storage, lower firmness throughout storage, followed by AEO 200 (1.937 kg/cm2) pears. Significant differences (p < 0.05) were also recorded between the applied treatments. The highest firmness was found in SEO 300 (2.957 kg/cm2) followed by SEO 250 (2.651 kg/cm2) and AEO 300 (2.549 kg/cm2). Here, there also were differences between the type and concentrations of EO used.

3.4. Dry Matter (DM)

Before the post-harvest treatments, the DM content was 15.04% and after treatments the DM content increased slightly without significant differences due to the presence of EOs on the pear surface. The results are in line with those found by Travers et al. (2014) [50].
After treatments, DM increased constantly throughout storage due to water losses through evapotranspiration, with significant differences (p < 0.05) between variants (Figure 5).
Surprisingly, the highest increase in DM was registered with the AEO 200 (25.6%) variant followed by the G 2.5 (23.57%) and control (23.51%) variants. Regarding the differences between the applied treatments, it can be observed that there are significant differences both in terms of the type and concentration of EO used.

3.5. Total Soluble Solids

Before the application of the post-harvest treatments, TSS was 11.42% in line with that found by Li et al. (2013) [51]. After the application of the EO treatments, TSS slightly increased, the highest values being found in the AEO 200 (12.43%) and G 2.5 (12.33%) variants (Figure 6). During storage, TSS increased constantly, the highest increase being found in the second month of storage. At the end of storage, TSS varied between 17.26% (control) and 13.80% (SEO 200).
The largest increases in TSS were observed in the second month of storage. Significant differences (p < 0.05) were found between the treatments applied throughout storage. Thus, the highest increase in TSS was found in the control variant (17.26%) followed by G 2.5 (16.73%) and AEO 300 (16.45%). The type of EO used and its concentration also influenced the TSS variation during storage. The lowest TSS value was found for the SEO 200 (13.80%) treated variants followed by AEO 200 (14.06%) and AEO 300 (14.06%).

3.6. Total Phenolic Content

Before the EO treatments, the TP was 56.25 mg GAE/100 g f.w. (Figure 7) which is consistent with the data found by Savic et al. (2021) [52] and He et al. (2022) [28] and lower than that found by Saoudi, Khennouf and Mayouf (2020) [53]. After one month of storage TP decreased then increased in the next two months with significant differences (p < 0.05) between variants. Significant differences (p < 0.05) were found at the end of storage for the variants studied. Thus, the highest TP was found in the SEO 300 treated variant (150 mg GAE/100 g f.w.) and the lowest in the SEO 200 treated variant (90.93 mg GAE/100 g f.w.).

3.7. Antioxidant Activity

Before the application of the post-harvest treatments, pear fruit had a relatively low free radical neutralization percentage (8.67%) compared with other species and data found by other authors [54].
Immediately after the application of the treatments, the percentage of free radical neutralization increased significantly in the EO-treated variants (Figure 8). This increment was a function of the type of EO used and proportional to the oil concentration.
After the first month of storage the percentage of free radical neutralization decreased with significant differences (p < 0.05) between variants, except for the control (13.20%) and G 2.5 variants (17.09%). In the last 2 months of storage, the percentage of free radical neutralization increased significantly, especially in the SEO treated variants except SEO 300. The highest percentage of free radical neutralization at the end of storage was found in SEO 250 (31.59%) and AEO 300 (31.11%). The lower concentrations of EO used resulted in a lower percentage of free radical neutralization even than the control (26.87%) and the G 2.5 variant (30.41%). An exception was found in the SEO 300 variant where the percentage of the neutralization of free radicals presented lower values than the SEO variants with lower concentrations (19.54%).

3.8. Color Evaluation

During storage, pears change color, which is an index to evaluate the ripeness of the fruit that can determine, along with other indexes, when the fruit is to be taken out of storage [55]. The color variation of pears during storage can be evaluated with the Yellowness index (14.86 b*/L*) and yellow color value (a*/b*). The data found on the Yellowness index (Figure 9) and yellow color value (Figure 10) shows that since the p-value of the F-test is greater than or equal to 0.05, there is not a statistically significant difference between the means of the eight variables at the 5% significance level for storage period. Significant differences (p < 0.05) were found only at the beginning and the end of storage. Similar results were found by Peralta-Ruiz et al. (2021) [55].
Prior to treatment, the yellowness index was 84.05, changing insignificantly after treatment. During storage, the yellowness index increased with significant differences between variants at the end of storage. The highest yellowness index value was found in the control (98.11), and the lowest values in the AEO 300 (88.22) and AEO 200 (89.14) variants. No significant differences (p < 0.05) were found between the variants analyzed in terms of the effect of the treatments applied. The same trend was found in yellow color value (Figure 10).

3.9. Total Aerobic Mesophilic Bacteria

Fungi were isolated from the surface of the examined pears. These were classified into the yeast category and did not register logarithmic growth during the storage period. No molds were present. The variants in which essential oils were used showed no fungi, providing evidence that no deterioration occurred during the storage period.
Before applying the treatments, TAMB was 5.07 log CFU/g. TAMB decreased immediately after applying the treatments with significant differences (p < 0.05) depending on the type and the concentration of the EO used (Figure 11).
The lowest TAMB was found in the SEO 300 (3.00 log CFU/g) variant, followed by AEO 300 (3.30 log CFU/g) and AEO 200 (3.60 log CFU/g). During storage, TAMB increased constantly with significant differences both between the storage periods and between the treatments applied. Regarding the differences between the types of EOs used, it was found that immediately after treatment, SEO had higher antimicrobial activity (p < 0.05) than AEO (3.00 for SEO 300, 3.30 for AEO 300, respectively). These significant differences (p < 0.05) were persistent throughout the storage period. The concentration of EO used influences the antimicrobial activity of the post-harvest treatment applied, with higher concentrations of EO leading to a more pronounced decrease in TAMB immediately after treatment (4.05 for SEO 200; 3.77 for SEO 250, 3.00 for SEO 300, respectively).

3.10. Sensory Analysis

The taste of the pear at the end of storage was rated by volunteers with scores between 3.4 (control) and 9.6 (SEO 300). In terms of color, the lowest scores were recorded for the control (2.7) and G 2.5 (4.9) variants, with significant differences from the EO treated variants which received the highest scores (9.5 for SEO 200 and AEO 300). The same trend was observed in terms of general acceptance, where the lowest scores were acquired by the control (3.8) and G 2.5 (5.7) variants and the SEO 300 (9.5) and SEO 250 (9.3) variants recorded the highest score (Figure 12).

4. Discussion

The gas chromatography analysis of anise essential oil has shown that the major component present is anethole. The gas chromatography analysis of sage essential oil revealed a different composition, with its own unique set of volatile compounds. Weight loss is a physiological indicator of the quality of a vegetal product during storage [46]. It was found that spraying pear fruit with SEO had a stronger effect on weight loss during storage compared to AEO. Also, the concentrations of the EO used influence weight loss by decreasing inversely in proportion to the concentration of the EO applied. A concentration of 300 ppm AEO thus had a similar effect on the weight loss of pears with a concentration of 200 ppm SEO. The possible reason behind the reduced weight loss in the treated fruits can be given by the presence of the essential oil combined with glycerin on the fruit surface, which on the one hand creates a semi-permeable layer on the fruit surface [56] and on the other hand, the essential oils used exhibit antimicrobial activity and the glycerin confers protection against the cold [57]. The high values of weight loss during the storage of the pears for 3 months lead to the idea that the storage time was too long in the control and glycerin-treated variants, resulting in a low level of acceptability.
It was found that higher concentrations of EOs maintain better fruit firmness and delay fruit ripening during storage. SEO was also found to perform better than AEO. AEO 300 showed similar action to SEO 250 in terms of pear firmness. The decrease in firmness is associated with the transformation of insoluble pectin in the fruit into soluble forms under the influence of protopectinase and pectinmethyl esterase because of ripening [58]. Therefore, the normal transformation of insoluble pectin into soluble pectin during storage appears to have been significantly delayed in both the SEO and AEO treated variants.
DM is an index of fruit ripening and quality. Measured at harvest, it can be used to predict the internal quality of pears after long periods of cold storage as has been used for apples [59,60]. DM is also identified as an indicator of eating quality [60]. Thus, the lowest DM values after storage were recorded for the SEO variants compared to the AEO variants, the lowest value being recorded for the SEO 300 variant. Also, the concentration of the EO used influenced the variation in the DM content, which decreased inversely proportional to the concentration of the EO used. At the end of storage, the lowest DM content was recorded for SEO 300 (16.54%) followed by the AEO 300 (21.47%) and AEO 250 variants (21.95%). The lower value of DM found in the variants treated with sage essential oil shows that this oil keeps water in the products during storage in a greater proportion compared to anise essential oil.
Higher EO concentrations resulted in increased TSS values for both types of EOs used. Still, the increase of TSS during storage agreed with the findings of Hussein et al. (2020) [61] and Al-Dairi & Pathare (2021) [62]. The increase in TSS during storage is due to the solubilization of starch with its transformation into soluble saccharides due to the continued ripening of the fruit [62].
Phenolic compounds are a category of secondary metabolites present in plants and are an integral part of human nutrition [53]. Immediately after the application of the post-harvest treatment, TP increased directly proportionally to the concentration of the EO used, probably due to oils adhering to the fruit surface. Differences were observed between the treatments applied. Thus, immediately after the application of the treatment, the highest TP value was found in the SEO treated variants followed by AEO. The increase in TP after post-harvest treatments is due to the polyphenol content of the EO used, AEO having the lowest TP content compared to SEO [63]. Differences between the EO concentrations used were also maintained.
Various herbal essential oils have strong antioxidant effects with beneficial effects on health, which have been reported in various studies [64]. The antioxidants protect cells from oxidative degradation by scavenging any free radicals [65]. The treatments performed with different EOs in different concentrations led to different behavior of the pear fruit during storage in terms of antioxidant activity, some authors [66], finding a downward trend of antioxidant activity in the pear epidermis during storage.
The pear is a fruit that continues to ripen after harvest. It is found that the treatments applied delay the ripening process of pears during storage. This delay may be due to protecting the polyphenolic compounds present on the pear’s surface by spraying [67]. The value obtained for the control for the yellowness index (close to 100) and yellow color value shows that the storage time was too long for this variant and the fruit over ripened. Moreover, physiological changes (spots) appeared on the skin of the pear in this variant, which are not accepted by consumers.
The microflora on raw vegetables and fruits is represented by spoilage bacteria, yeasts and molds but also occasional pathogenic bacteria and viruses capable of causing human infections [39]. The TAMB count on the surface of the fruits is one of the microbiological indicators of their quality. TAMB reflects the existence of favorable conditions for the multiplication of microorganisms [68].
Bacteria of the genus Bacillus, Erwinia and Pseudomonas as well as fungi of the genus Penicillium, Botritys and Mucor were isolated from the surface of the pear analyzed before treatment with sage and aniseed essential oils. After the application of EO treatments, there was a reduction in the number of microorganisms but also a change in the type of microorganisms responsible for pear fruit spoilage. Of the bacteria, the genus Pseudomonas and Erwinia were found throughout the storage period, while Bacillus was inhibited in a higher proportion. This can be explained by the lower resistance of G (+) bacteria compared to G (−) bacteria to the action of the compounds contained in the tested oils.
Among the molds, the Penicillium and Botritys genus persisted during the storage period, but the number of these fungi was diminished immediately after treatment. The multiplication of these fungi was carried out by the viable remaining spores. The two mold genera are frequently isolated from pear and other pectin-rich fruits [69,70].
There were significant differences in the volunteer response to both the types and concentrations of the EOs used. In terms of the differences between the types of EO used, the volunteers appreciated the pears treated with sage essential oil more than those treated with aniseed essential oil. As a result of the sensory evaluation, it was determined that the volunteers scored the SEO 300 and SEO 250 variants the highest. As for the aniseed essential oil-treated variants, although the volunteers appreciated the color of the fruit, they gave lower scores in terms of the taste and general acceptance of the pear fruit.

5. Conclusions

Spraying pear fruits with aqueous solutions containing glycerol (2.5%) and various concentrations of sage and aniseed EOs results in an extension of the pears’ shelf life.
Furthermore, these treatments also contributed to a more modest decline in pear firmness during the storage period, with SEO 300 recording 2.957 kg/cm² compared to 1.529 kg/cm² in the control treatment at the end of storage. Additionally, the use of essential oils led to a reduced weight loss during storage. This method also reduces the breakdown of phenolic compounds in fully mature pears, thereby preserving the pears’ antioxidant properties.
The application of essential oil sprays on pears can effectively prolong their shelf life by as much as 30 days when stored at 3 ± 1 °C with a relative humidity of 80%. The specific extension in shelf life depends on the type and concentration of the essential oil utilized.
The most effective essential oil is found to be sage essential oil in concentrations of 250 and 300 ppm. For anise essential oil, the most effective concentration was 300 ppm.
EO concentrations higher than 250 ppm decreased the TAMB immediately after treatment (p ≤ 0.05) and the differences persisted until the end of storage.
The control and G 2.5 groups exhibited consistent increases in the total count of aerobic mesophilic bacteria (TAMB) throughout the storage duration. By the end of the storage period, TAMB levels in these groups had doubled in comparison to the EO-treated variations (reaching 6.78 in the control and 3.90 in ESO 300).
The use of post-harvest treatments by spraying pear fruits with aqueous solutions of glycerol and essential plant oils can increase shelf life by reducing microbiological load on the fruit surface, modifying evapotranspiration and slowing fruit ripening.
Sage essential oil was found to be more effective than aniseed essential oil and can be used in concentrations of 250 ppm.

Author Contributions

M.E.I.: project administration, writing—original draft, investigation, statistical data processing. F.T.: writing—original draft, and review and editing, investigation. A.B.: investigation, writing—original draft. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Informed consent was obtained from all subjects involved in the study.

Data Availability Statement

Data are contained within the article.

Acknowledgments

We thank engineer Carmen Andrei for administrative and technical support.

Conflicts of Interest

The authors declare no conflict of interest.

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Coatings 14 00010 g001
Figure 1. Gas chromatogram of the Pimpinella anisum essential oil extract.
Figure 1. Gas chromatogram of the Pimpinella anisum essential oil extract.
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Figure 2. Gas chromatogram of the Salvia officinalis essential oil extract.
Figure 2. Gas chromatogram of the Salvia officinalis essential oil extract.
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Figure 3. Effects of the applied treatments on weight loss (%) during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
Figure 3. Effects of the applied treatments on weight loss (%) during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
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Figure 4. Effects of the applied treatments on fruit firmness (kg/cm2) during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
Figure 4. Effects of the applied treatments on fruit firmness (kg/cm2) during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
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Figure 5. Effects of the applied treatments on DM content (%) during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
Figure 5. Effects of the applied treatments on DM content (%) during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
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Figure 6. Effects of the applied treatments on Total soluble solids content (%) during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
Figure 6. Effects of the applied treatments on Total soluble solids content (%) during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
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Figure 7. Effects of the applied treatments on total phenolic content (mg GAE/100 g f.w.) during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
Figure 7. Effects of the applied treatments on total phenolic content (mg GAE/100 g f.w.) during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
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Figure 8. Effects of the applied treatments on antioxidant activity (%) during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
Figure 8. Effects of the applied treatments on antioxidant activity (%) during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
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Figure 9. Effects of the applied treatments on the yellowness index during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
Figure 9. Effects of the applied treatments on the yellowness index during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
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Figure 10. Effects of the applied treatments on the yellowness color value during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
Figure 10. Effects of the applied treatments on the yellowness color value during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
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Figure 11. Effects of the applied treatments on the total aerobic mesophilic bacteria during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
Figure 11. Effects of the applied treatments on the total aerobic mesophilic bacteria during storage for 3 months at 3 ± 1 °C and 80% relative humidity. Bars sharing the same sampling time and denoted by distinct lowercase letters exhibit statistically significant differences, as determined by the least significant difference (LSD) test (p < 0.05). Similarly, bars representing the same treatment across different storage times and marked with varying uppercase letters signify significant differences, as determined by the LSD test (p < 0.05).
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Figure 12. Sensory analysis of pears after storage.
Figure 12. Sensory analysis of pears after storage.
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Table 1. Type and concentrations of EO used for pear spraying.
Table 1. Type and concentrations of EO used for pear spraying.
VariantThe Treatment Used
Controluntreated
G 2.5aqueous glycerol solution 2.5%
ESO 200200 ppm essential sage oil in aqueous glycerol solution
ESO 250250 ppm essential sage oil
ESO 300300 ppm essential sage oil
AEO 200200 ppm anise essential oil
AEO 250250 ppm anise essential oil
AEO 300300 ppm anise essential oil
Table 2. GC-MS analysis of the Pimpinella anisum and Salvia officinalis essential oil extract.
Table 2. GC-MS analysis of the Pimpinella anisum and Salvia officinalis essential oil extract.
No.RTCompoundAnise EO (%)Sage EO (%)
111.95Thujene-0.572
212.46α-Pinene-1.878
313.38Camphene-3.169
414.32Sabinene-0.482
514.48β-Myrcene-1.535
614.63β-Pinene-5.838
715.53Morillol-0.388
816.26α-Terpinene-0.511
916.58trans-Ocimene-0.184
1017.24β-Phellandrene-0.170
1117.64Eucalyptol-17.615
1218.33γ-Terpinene-0.857
1320.09Limonene0.0051.903
1420.42o-Cymene0.0080.333
1521.06Sabinene hydrate-0.185
1621.77Terpinolene0.0030.407
1723.21trans-Sabinene hydrate-0.164
1824.05Thujone-21.996
1924.64β-Thujone-5.076
2025.49Isothujol-0.109
2125.74cis-Sabinol-0.207
2226.17Linalool0.0030.240
2326.77L-Fenchone0.101-
2426.87Camphor-23.846
2527.22Terpinen-4-ol-1.348
2627.71endo-Borneol-0.898
2728.35α-Terpineol-0.350
2832.79Bornyl acetate-0.160
2933.04Estragole0.418-
3038.1Carvone0.003-
3138.99Caryophyllene-1.119
3239.61Anethole97.467-
3339.66Aromandendrene-0.125
3440.49Anisaldehyde0.536-
3540.66γ-Muurolene-2.540
3641.88Ledene-0.105
3742.76β-Cadinene-0.133
3844.7β-Elemene0.007-
3948.18Viridiflorol-5.056
4048.84Anisylacetone0.011-
4149.3Humulene-1,2-epoxide-0.502
4250.26Curcumene0.087-
4350.49Zingiberene0.094-
4450.72γ-Himachalane1.102-
4551.28α-Longipinene0.070-
4651.98β-Himachalene0.032-
4753.814,5-dehydro-Isolongifolene0.018-
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MDPI and ACS Style

Ionica, M.E.; Bita, A.; Tutulescu, F. Using the Essential Oils of Sage and Anise to Enhance the Shelf Life of the Williams (sin. Bartlett) Pear. Coatings 2024, 14, 10. https://doi.org/10.3390/coatings14010010

AMA Style

Ionica ME, Bita A, Tutulescu F. Using the Essential Oils of Sage and Anise to Enhance the Shelf Life of the Williams (sin. Bartlett) Pear. Coatings. 2024; 14(1):10. https://doi.org/10.3390/coatings14010010

Chicago/Turabian Style

Ionica, Mira Elena, Andrei Bita, and Felicia Tutulescu. 2024. "Using the Essential Oils of Sage and Anise to Enhance the Shelf Life of the Williams (sin. Bartlett) Pear" Coatings 14, no. 1: 10. https://doi.org/10.3390/coatings14010010

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