The use of volatile antimicrobial emitting sachets for extending the shelf-life of packaged sweetbread

Abstract

This study investigated the use of active sachets that emit volatile antimicrobial compounds — ethanol, with and without the addition of cinnamon oil (CO) — to extend the shelf-life of packaged sweetbread. The addition of CO was expected to mask any undesirable odor from ethanol and enhance its antimicrobial efficacy. The active sachets contained ethanol emitter (EE) only and EE combined with CO (ECOE) at the dose of 0.5 and 1 times the minimum inhibitory concentration (MIC). The effects of active sachets were evaluated based on mold (Rhizopus stolonifer) growth inhibition, hardness, and sensory properties of the packaged sweetbread. The obtained results showed that the mold growth fully covered the Petri dish containing potato dextrose agar (PDA) in 3 days and 4 days in samples without an active sachet and with the EE sachet, respectively. Interestingly, there was no mold growth in samples containing active sachets ECOE 0.5 MIC and 1 MIC. The hardness of the sweetbread, measured with a texture analyzer, was 2105.2 gf for the control, 1375.0 gf for EE, 1313.0 gf for ECOE 0.5 MIC, and 1224.8 gf for ECOE 1 MIC, thus indicating a slower staling rate when using active sachets. Sensory evaluation revealed that there was no significant difference (p > 0.05) in the overall acceptance parameter between sweetbread packaged with and without active sachets (EE and ECOE) treatments indicating there was no decreasing consumer preference. The addition of CO to ethanol emitters enhanced the extension of the shelf-life of packaged sweetbread by inhibiting mold growth and reducing staling without decreasing sensory quality.

Keywords:  Ethanol emitter; Antimicrobial packaging; Controlled-release; Active packaging; Active sachets; Cinnamon oil

HIGHLIGHTS

The application of an ethanol emitter combined with cinnamon oil may serve as an antimicrobial active packaging solution to prolong the shelf-life of sweetbread

The combination of ethanol with cinnamon oil more effectively inhibits the growth of Rhizopus stolonifer compared to the use of ethanol alone

The combination of ethanol and cinnamon oil emitter effectively reduces the staling rate and enhances overall acceptance among panelists

1 Introduction

Active packaging is related to packaging that is treated with active ingredients using certain principles. Active packaging is superior to conventional packaging because it is more effective in maintaining food quality and increasing shelf-life (Quintavalla & Vicini, 2002). Ethanol emitters are active packaging with a release principle where ethanol is the active ingredient released into the headspace of the food packaging (Latou et al., 2010). Ethanol was reported to inhibit the growth of microorganisms in food thus extending the shelf-life of food products (Janjarasskul et al., 2016; Zaitoon et al., 2022).

Controlled-release is required for ethanol emitters so that ethanol as an active ingredient does not evaporate quickly due to its high evaporation rate (Kita et al., 2018; Mu et al., 2017). The use of carrier material will affect the ability of active ingredients to be released into food packaging (Otoni et al., 2016). Silicon dioxide, zeolite, PS/EVOH/PE tray, and PVDC/nylon/LDPE have been used in previous research as carrier materials for ethanol (Hempel et al., 2013; Mu et al., 2017; Smith et al., 1987, 1995). The high evaporation rate of ethanol can be detrimental due to the undesirable aroma of ethanol in food products. It is also possible to add capping agents to ethanol to cover up its smell. Cinnamon oil was utilized in this investigation; it has a pleasant aroma and is said to have strong antibacterial properties (Clemente et al., 2019; Souza et al., 2013). The addition of cinnamon oil to the ethanol emitter can increase the antimicrobial ability of the active sachet. Figure 1 illustrates the use of controlled-release ethanol emitter with cinnamon oil on food products. The presence of antimicrobials in active packaging will prolong the lag phase so that the growth rate is reduced and ultimately reduces the live cells of microbes (Fadiji et al., 2023).

Figure 1
Illustration of controlled-release ethanol emitter with cinnamon oil on food products.

Bread has a short shelf-life or is categorized as perishable food. Damage to bread includes physical, chemical, and microbiological damage. Physical damage to bread is also known as staling. Staling occurs due to starch, gluten, and water in bread experiencing retrogradation and dehydration so that the bread becomes hardened (Curti et al., 2014). A rancid aroma in bread can characterize chemical damage to bread. Microbiological damage to bread is mainly caused by mold growth, especially Rhizopus stolonifer (Natawijaya et al., 2015; Tiyas et al., 2023). Damage to bread can be prevented by providing controlled-release ethanol emitters added with cinnamon oil. The use of controlled-release ethanol emitters with cinnamon oil is expected to extend the shelf-life of bread without significantly imparting its aroma characteristics. This research aims to develop controlled-release ethanol emitter with cinnamon oil active packaging and its application to extend the shelf-life of bread products. Specifically, this study aims to look at the anti-mold ability of controlled-release ethanol emitter with cinnamon oil and to look at changes in the physical and sensory properties of bread packaged with the addition of controlled-release ethanol emitter with cinnamon oil sachets.

2 Material and methods

2.1 Materials

The materials used include active ingredients for making ethanol emitter active packaging in the form of ethanol (Merck, food grade), silica dioxide (Darisa, food grade), sodium stearate (Techno Pharmachem, food grade), cinnamon oil (Symrise Asia Pacific Pte Ltd, food grade), as well as sachets of Mg paper/PE, and sweetbread. Materials for anti-mold testing were potato dextrose agar (PDA) media, sweetbread, controlled-release ethanol emitter with cinnamon oil sachets, and Rhizopus stolonifer isolates.

2.2 Preparation of controlled-release ethanol emitter with cinnamon oil sachets

The controlled-release ethanol emitter with cinnamon oil was based on Mu et al. (2017) with modifications. The ethanol emitter formulation consisted of ethanol 53.7% (v/v), sodium stearate 2.6% (v/v), silicon dioxide 34% (v/v) and distilled water 9.7% (v/v). The treatment formulation added cinnamon oil as much as 0.5 MIC and 1 MIC. The active packaging was prepared by mixing ethanol, sodium stearate, distilled water, and cinnamon oil. The solution was heated to 70 ^oC and then combined with SiO₂, and the mixture was obtained in powder form. The powder obtained was weighed and packaged in MG paper/PE measuring 2.5 cm x 4 cm for anti-mold ability analysis and 5 cm x 4 cm for staling rate and sensory analysis, then sealed.

2.3 Anti-Mold ability analysis (Krisch et al., 2013)

The analysis was performed by spot-inoculating PDA media using a 10⁴ spore/mL Rhizopus stolonifer mold suspension in a petri dish. The diameter of the Petri dish was 8.5 cm. Sachets of controlled-release ethanol emitter with cinnamon oil were attached to the lid of the Petri dish and incubated for four days. Observations were made by measuring the diameter of molds that grew on PDA media and expressed as a percentage of mold growth.

2.4 Texture analysis of sweetbread (AACC, 1999; Jensen et al., 2015)

Texture analysis was conducted to determine the staling rate of bread. Hardness values on texture analysis could represent the staling rate of sweetbreads after storage with controlled-release ethanol emitters with cinnamon oil. Texture analysis was performed using TA-XT2i (stable microsystem, UK) based on (AACC, 1999; Jensen et al., 2015). The AACC 74-09.01 method was performed by preparing an 85 mm piece of bread; the probe was 35 mm thick. The instrument was set with a pre-test speed of 2.00 mm/s, crosshead speed of 100 mm/min, chart speed of 500 mm/min, post-test speed of 10.0 mm/s, rupture test distance of 1 mm, and 40% compression. The test results were used to measure the hardness value. The texture of the bread was analyzed on storage days 0, 3, 7, and 14.

2.5 Sensory analysis of sweetbread (Meilgaard et al., 2016)

Sensory analysis was conducted using an attribute rating test and panelist acceptance test of sweetbread based on Meilgaard et al. (2016). The panelists used were untrained panelists with an age range of 17-40 years. The attributes tested were the intensity aroma of ethanol, the intensity aroma of cinnamon oil, and overall panelist acceptance of the sweetbread with a rating of 0 to 10. Sensory tests were conducted on bread stored on days 0, 1, 3, and 7.

2.6 Statistical analysis

Data analysis was conducted using Excel 2016 and IBM SPSS v.25 software. Data processed using Excel software included the anti-mold ability of controlled-release ethanol emitter with cinnamon oil sachets. Texture and sweetbread sensory data were processed using randomized factorial design Analysis of Variance (ANOVA) with the help of IBM SPSS v.25 software.

3 Result and discussion

3.1 Mold growth inhibitory of the controlled-release ethanol emitter and cinnamon oil

The Figure 2 depicts the inhibition of Rhizopus stolonifer mold growth on PDA media by different treatments, including a control (blue), an ethanol emitter (EE, orange), and two concentrations of a controlled-release ethanol emitter combined with cinnamon oil extract (ECOE 0.5 MIC and ECOE 1 MIC). The mold growth inhibition was evaluated by measuring the diameter of the mold colony for 4 days with the maximum possible growth equal to the Petri dish diameter (8.5 cm). In the control Petri dish, mold growth increased rapidly, reaching full coverage (8.5 cm) by day 3. The active sachet EE only showed inhibition activity, but mold growth still occurred until 100% after 4 days. While the ECOE treatments (0.5 MIC and 1 MIC) could totally inhibit mold growth indicated by no visible mold growth after 4 days.

Figure 2
Diameter of Rhizopus stolonifer mold growth on Petri dish containing PDA media (D = 8.5 cm)

The EE has effectively delayed the growth of mold, particularly in the early days (up to day 2), but mold eventually overcomes the inhibition by day 4, reaching full growth. This is in agreement with the result that was reported by Berni & Scaramuzza (2013). The active sachets that emit ethanol with cinnamon oil (ECOE 0.5 MIC and 1 MIC) demonstrated a complete inhibition of mold growth for 4 days. These results indicate that the controlled-release ethanol emitter in combination with cinnamon oil (ECOE) is a potent antifungal treatment against Rhizopus stolonifer. Cinnamon oil was reported to have broad-spectrum antimicrobial activity and is a strong antimicrobial compared to other essential oils (Kowalska et al., 2021). Moreover, Hurley et al. (2013) reported that the addition of cinnamon oil in the packaging of bakery products avoid mold growth until seven days of storage.

Ethanol produced antimicrobial activity that was effective against molds and inhibited the growth of yeasts and bacteria (Kumar et al., 2018). Spraying 95% ethanol at a concentration of 0.5% to 1.5% (w/w) could inhibit mold growth and increase the shelf-life of bread products (Kumar et al., 2018). Ethanol has been shown to inhibit bacteria on the surface of fish fillets (Olsen, 2021). Cinnamon oil used as a capping agent has antimicrobial activity that is effective against Gram-positive and Gram-negative bacteria, mold, and yeast and inhibits toxin production in molds (Gupta et al., 2008; Haddi et al., 2017; Suppakul, 2016). The phase of mold growth affected by antimicrobials is the lag phase, where the mold is still adjusting to the surrounding environment (Tapía et al., 2020). The addition of cinnamon oil to ethanol emitters provided a synergistic antimicrobial effect so that the antimicrobial strength of ECOE 0.5 MIC and ECOE 1 MIC sachets was higher than EE sachets against Rhizopus stolonifer.

3.2 Staling rate analysis of bread stored with controlled-release ethanol emitter with cinnamon oil

The Figure 3 represents an analysis of sweetbread staling, measured by hardness, over a storage period of 0, 3, 7, and 14 days. The sweetbread was subjected to different treatments, including a control (without treatment), EE, and ECOE) at two different concentrations: 0.5 MIC and 1 MIC. The control sample (blue line) showed the highest rate of staling, with the hardness reaching approximately 2200 by day 14. The linear regression equation for the control group (y = 108.12x + 662.43, 𝑅² = 0.7686) suggests a strong positive correlation between storage time and hardness. The higher slope (108.12) indicates the most rapid staling rate among the treatments. The sweetbread treated with ethanol emitter only, (EE, orange line) showed a relatively lower staling rate compared to the control, with a final hardness below 1500 on day 14. The regression equation (y = 66.551x + 498.55, 𝑅² = 0.8327) indicates a lower staling rate (slope = 66.55) than the control, suggesting that EE slows down the staling process. The sweetbread treated with a lower concentration of cinnamon oil extract (0.5 MIC, grey line) had a hardness level that increased steadily, reaching around 1700 by day 14. The regression equation (y = 71.005x + 523.66, 𝑅2 = 0.6896) showed a slightly higher staling rate (slope = 71.01) than EE but lower than the control. The highest concentration of cinnamon oil extract (1 MIC, yellow line) showed the lowest overall staling effect, with the final hardness remaining below 1300 by day 14. The regression equation (y = 54.361x + 559.61, 𝑅² = 0.677) showed the slowest staling rate (slope = 54.36) among all treatments. The results demonstrate that the use of a controlled-release ethanol emitter in combination with cinnamon oil, particularly at 1 MIC, significantly slows down the staling rate of sweetbread during storage.

Figure 3
Interaction plot of sweetbread hardness with storage time on days 0, 3, 7, and 14 with control, EE, ECOE (0.5 MIC), ECOE (1 MIC) treatments.

Bread hardness increases with the length of storage due to moisture loss and starch retrogradation during storage (Ammar et al., 2016). The hardness value is strongly related to bread staling. Staling in bread occurs due to physical and chemical changes in bread, such as crumb hardening, crust softening, and loss of fresh bread flavor (Abd-El-Khalek et al., 2019). Staling is closely related to starch retrogradation and water transfer from the inside of the bread to the outside. EE, ECOE (0.5 MIC), and ECOE (1 MIC) sachets had low hardness values, so they got stale quickly when compared to the control. It is presumably due to the water contained in the active packaging, which is more accessible to evaporate first, so the water contained in the bread, which is bound, is retained longer. Besides, the emitted cinnamon oil might play an important role as a plasticizer for the bread keeping the freshness while increasing its antimicrobial capacity.

3.3 Sensory analysis

The result of sensory analysis (Table 1) presents the evaluation of bread stored with a controlled-release ethanol emitter combined with cinnamon oil under different treatments over 7 days. The sensory parameters analyzed include Aroma of Ethanol, Aroma of Cinnamon Oil, and Overall Acceptance. The treatments consisted of a control, EE, and ECOE at two concentrations: 0.5 MIC and 1 MIC. Each parameter was assessed on storage days 0, 1, 3, and 7. Statistical differences between the treatments are indicated by different letters (uppercase across rows for storage days, lowercase across columns for treatments) based on Duncan’s Multiple Range Test (DMRT).

Table 1
Sensory analysis of bread stored with controlled-release ethanol emitter with cinnamon oil.

Sensory Parameters	Treatments	Days of Storage
		0	1	3	7
Aroma of Ethanol	Control	0.97Aa	1.09Aa	0.76Aa	0.91Aa
	EE	0.97Aa	1.18Aa	1.27Aa	1.64Aa
	ECOE (0.5 MIC)	0.97Aa	2.00Bab	1.30Ba	1.94Bab
	ECOE (1 MIC)	0.97Aa	2.67Bb	1.55ABa	2.27Bab
Aroma of Cinnamon Oil	Control	1.18Aa	1.61Aa	1.36Aa	0.88Aa
	EE	1.18Aa	4.15Bb	4.24Bb	4.15Bb
	ECOE (0.5 MIC)	1.18Aa	5.33Bc	5.09Bb	4.85Bb
	ECOE (1 MIC)	1.18Aa	5.82Bc	5.21Bb	4.79Bb
Overall Acceptance	Control	8.30Aa	7.55Aa	6.85Ba	5.94Ba
	EE	8.30Aa	7.58Aa	6.70ABa	6.15Ba
	ECOE (0.5 MIC)	8.30Aa	7.00Ba	7.21Ba	6.36Ba
	ECOE (1 MIC)	8.30Aa	6.91Ba	6.88Ba	6.76Ba

• Notes: numbers in the same row followed by uppercase letters indicate significant difference at the level α = 0.05; and in the same column numbers followed by different lowercase letters indicate significant difference at the level α = 0.05 by Duncan Multiple Range Test (DMRT).

The aroma of ethanol attribute of samples with control and EE treatments had no simple effect in the storage on days 0, 1, 3, and 7, while the ECOE 0.5 MIC and ECOE 1 MIC treatments had a simple effect in the storage on days 0, 1, 3, and 7. Storage on days 0 and 3 had no simple effect on the aroma of the ethanol attribute, while storage on days 1 and 7 had a simple effect on the aroma of the ethanol attribute. The intensity of the aroma of ethanol in the samples was more vigorous in the ECOE 0.5 MIC and ECOE 1 MIC treatments. The aroma of cinnamon oil in each treatment had a simple effect on days 1, 3, and 7 compared to the control. The smell of cinnamon oil by panelists in the EE treatment was thought to be due to being distracted by the reference to cinnamon oil that was smelled first. Overall acceptance by panelists in EE, ECOE 0.5 MIC, and ECOE 1 MIC was not significantly different but significantly different when viewed from storage days 0, 1, 3, and 7.

Sensory attribute rating assessment is included in sensory analysis where panelists are asked to give intensity ratings to selected attributes on a numerical intensity scale (Meilgaard et al., 2016). The addition of ethanol and cinnamon oil, which has a distinctive flavor and aroma, will affect the sensory aspect of the product depending on the concentration (Brnawi et al., 2018). Testing the intensity of ethanol aroma and cinnamon oil aroma is accompanied by consumer acceptance to see whether the presence of ethanol aroma and cinnamon oil aroma at the tested concentration is still acceptable to panelists. The result of sensory analysis showed that panelists still accepted the aroma of EE, ECOE 0.5 MIC, and ECOE 1 MIC, even though there is a reasonably strong aroma of ethanol and cinnamon oil.

4 Conclusion

The addition of cinnamon oil significantly enhanced the anti-mold activity of ethanol emitters. While the ethanol emitter (EE) alone showed limited effectiveness against Rhizopus stolonifer, both ECOE 0.5 MIC and ECOE 1 MIC demonstrated superior mold inhibition. Sweetbread stored with EE, ECOE 0.5 MIC, and ECOE 1 MIC exhibited lower hardness values compared to the control, indicating a reduced staling rate. Among the treatments, ECOE 1 MIC was the most effective at slowing the staling process. Sensory evaluations revealed that the presence of ethanol and cinnamon oil aromas in the treated bread did not negatively impact overall acceptance by the panelists. Overall, ECOE 0.5 MIC emerges as the most promising formulation, as it effectively inhibits mold growth, reduces staling and is well-accepted by consumers, all while using a lower concentration of cinnamon oil.

Acknowledgements

We are grateful to the Directorate of Research, Technology and Community Service, Ministry of Education, Culture, Research and Technology of Indonesia for funding this research through the programs Pendanaan Thesis Magister (PTM) 2023.

• Cite as:
  Shalita, A.,Adawiyah, D. R., & Suyatma, N. E. (2024). The use of volatile antimicrobial emitting sachets for extending the shelf-life of packaged sweetbread. Brazilian Journal of Food Technology, 27, e2024028. https://doi.org/10.1590/1981-6723.02824
• Funding:
  Ministry of Education and Culture Republic of Indonesia (PTM 2023)

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Edited by

• Associate Editor:
  Cassandra Dalle Mulle Santos.

Publication Dates

• Publication in this collection
  09 Dec 2024
• Date of issue
  2024

History

• Received
  24 Mar 2024
• Accepted
  16 Oct 2024