1. INTRODUCTION:

Herbal extracts have long been integral to human health, offering a wealth of therapeutic benefits that have been harnessed across cultures for centuries^1,2. Among the most versatile and widely utilized herbal remedies is Cinnamomum verum (Cinnamon), a plant renowned not only for its culinary uses but also for its potent pharmacological properties^3,4. Cinnamon has demonstrated remarkable biological activities, including antimicrobial⁵, anti-bacterial⁶, antioxidant^7,8, anti-inflammatory, and anticancer effects^9,10. These therapeutic effects are primarily attributed to its rich array of bioactive compounds, such as cinnamaldehyde, eugenol, and polyphenols, which have been widely studied for their potential in modern medicine^11,12.

Fig. 01 Major phytochemical components of Cinnamon: (a) Cinnamaldehyde, (b) Euginol, (c) Linalool, (d) Cinnamyl acetate (e) β-Caryophyllene

While the therapeutic potential of cinnamon is well-established, the stability of its active constituents during processing and storage remains a critical, yet often overlooked, consideration. The evaporation of solvent during the preparation of herbal extracts, particularly ethanolic extracts, can lead to significant alterations in the chemical composition of these extracts, potentially diminishing their efficacy¹³. The impact of evaporation on phytochemical retention is influenced by multiple factors, including the geometry of the container and environmental conditions, yet this relationship is rarely explored in depth, especially for widely used herbs like cinnamon¹⁴. This study aims to bridge this gap by investigating the evaporation dynamics of ethanolic cinnamon extract under different container geometries and environmental conditions. By examining how surface area and environmental factors affect the rate of evaporation, this research delves into a novel aspect of herbal extract preservation^15,16. Using test tubes, conical flasks, petri plates, china dishes, and beakers as experimental setups, we measured the weight loss of the extract at intervals over 24 hours, while monitoring the changes in phytochemical composition through Thin-Layer Chromatography (TLC)¹⁷. The primary goal was to understand how these physical factors influence the preservation of key bioactive compounds in cinnamon extract, with the aim of improving its formulation and storage for optimal therapeutic efficacy. The findings from this study are poised to make a significant contribution to the field of natural product chemistry. By shedding light on the critical factors that influence the stability of herbal extracts, this research could inform best practices in the production, storage, and application of cinnamon-based products, not only in the pharmaceutical industry but also in the food and cosmetic sectors. Ultimately, this work underscores the importance of carefully controlling environmental and geometric factors to ensure that the valuable bioactive compounds in herbal extracts are preserved, maximizing their potential benefits to human health.

2. MATERIALS AND METHODS:

2.1 Materials

· Cinnamon (Cinnamomum verum): Dried cinnamon bark was obtained from local market.

· Ethanol: Analytical grade ethanol (95%) was used for extraction.

· Reagents for Thin-Layer Chromatography (TLC): Solvents and reagents for TLC, including silica gel plates and the appropriate mobile phases for phytochemical analysis.

2.2 Apparatus:

Evaporation Vessels: The evaporation study was carried out using various laboratory vessels with different geometries, including: test tubes, conical flasks, petri plates, china dishes, beakers.

· Analytical Balance: To measure the weight of the samples at specified time intervals.

· Incubator/Environmental Control: For controlled environmental conditions, an incubator was used to maintain temperature and humidity.

· Thin-Layer Chromatography (TLC)¹⁸ Setup: A standard TLC apparatus with UV light and a developing chamber was used for analyzing the phytochemical composition.

2.3 Preparation of Ethanolic Cinnamon Extract

Cinnamon bark was finely ground to a powder using a mortar and pestle. A 10g sample of the cinnamon powder was macerated in 100 mL of 95% ethanol in a sealed container for 48 hours with occasional shaking. The mixture was then filtered through Whatman filter paper, and the resulting ethanolic extract was concentrated by evaporating the solvent under reduced pressure using a rotary evaporator. (Figure-2).

Fig. 3. Open vs. Controlled evaporation study of herbal ethanolic extract (cinnamon)

Fig. 4. (a) TLC Chamber for standard mobile phase Hexane: Ethyl acetate (6:4, 7:3, 8:2) (b) Iodine Chamber (c) TLC plates of fresh ethanolic extract of cinnamon

Fig. 5. Open vs. Controlled TLC study of herbal ethanolic extract (cinnamon) kept in- (a) Beaker (b) Conical flask (c) Test tube (d) China dish (e) Petri plate

Table 1: Observation of evaporating behaviour of herbal ethanolic extract in open vessels

S. No.	Apparatus	Weight of Empty Apparatus	Initial weight (0 hr)	Weight after 1:00 hr	Weight after 2:00 hr	Weight after 6:00 hr	Weight after 12:00 hr	Weight after 18:00 hr	Weight after 24:00 hr
1.	Conical-flask	68.42 g	73.41 g	73.34 g	73.27 g	73.06 g	72.81 g	72.34 g	72.09 g
2.	Beaker	35.68 g	40.65 g	40.23 g	39.82 g	39.12 g	38.52 g	36.93 g	36.85 g
3.	Test-tube	16.39 g	21.31 g	21.29 g	21.25 g	21.08 g	20.91 g	20.79 g	20.67 g
4.	Petri-plate	38.17 g	43.06 g	39.86 g	39.12 g	38.23 g	38.23 g	38.23 g	38.23 g
5.	China-dish	92.79 g	97.75 g	95.50 g	93.96 g	93.75 g	93.11 g	93.11 g	93.11 g

Table 2: Observation of evaporating behaviour of herbal ethanolic extract in controlled vessels

S. No.	Apparatus	Weight of Empty Apparatus	Initial weight (0 hr)	Weight after 1:00 hr	Weight after 2:00 hr	Weight after 6:00 hr	Weight after 12:00 hr	Weight after 18:00 hr	Weight after 24:00 hr
1.	Conical-flask	67.12 g	72.05 g	72.04 g	72.04 g	72.03 g	71.96 g	71.88 g	71.83 g
2.	Beaker	33.97 g	38.95 g	38.94 g	38.92 g	38.92 g	38.85 g	38.74 g	38.67 g
3.	Test-tube	18.46 g	23.42 g	23.41 g	23.41 g	23.39 g	23.31 g	23.28 g	23.23 g
4.	Petri-plate	37.92 g	42.87 g	42.80 g	42.53 g	42.41 g	42.32 g	42.25 g	42.19 g
5.	China-dish	93.62 g	98.59 g	98.52 g	98.49 g	98.41 g	98.32 g	98. 25 g	98. 21 g

Among the tested mobile phase ratios, Hexane: Ethyl Acetate (7:3) demonstrated the most effective separation of the major constituents present in the ethanolic extract of cinnamon. This ratio provided well-resolved and distinct spots with R_f values falling within the ideal range of 0.3 to 0.7, ensuring accurate identification and comparative analysis. While the 8:2 ratio favoured the migration of non-polar compounds and the 6:4 ratio enhanced the movement of polar components, the 7:3 ratio offered a balanced polarity suitable for resolving both types of compounds efficiently as shown in fig. 05 (c). Therefore, based on its superior performance in achieving optimal separation and clarity of spots, the 7:3 Hexane: Ethyl Acetate system was selected as the most suitable mobile phase for further chromatographic studies.

· Open Conditions: In open conditions, a decrease in the intensity of the key bioactive compound spots was observed, particularly for cinnamaldehyde and eugenol. At the 24-hour mark, some compounds appeared to have been significantly reduced or were no longer detectable.

· Controlled Conditions: In the controlled environment, the intensity of the phytochemical spots remained relatively stable throughout the 24-hour period. This suggests that the controlled environment was more effective in preserving the bioactive compounds in the cinnamon extract.

Visual analysis of the TLC plates revealed that under open conditions, the intensity of key phytochemical spots, particularly for cinnamaldehyde and eugenol, declined markedly over time. At 24 hours, several spots showed faint or nearly undetectable intensities, suggesting significant degradation or volatilization. In contrast, TLC plates from controlled environments retained strong to moderate spot intensity, indicating better preservation. Future studies may incorporate densitometric scanning for quantitative estimation of compound degradation.

Fig. 08 Comparison of R_fValues of Cinnamon Extract in Open vs Controlled condition (24h TLC)

4. DISCUSSION:

The experiment demonstrated that apparatus geometry and environmental exposure significantly influence the stability of cinnamon ethanolic extracts. Extracts in open vessels, especially those with larger surface areas like petri plates and china dishes, exhibited notable reductions in R_f values and spot intensities after 24 hours, indicative of compound degradation or volatilization. In contrast, controlled conditions using aluminium foil coverings preserved the extract's phytochemical profile more effectively, as evidenced by consistently higher R_f values and stronger TLC spots. These findings underscore the importance of environmental control in herbal extract processing and storage to maintain phytochemical efficacy.

4.1 Interpretation of Evaporation Dynamics:

This study demonstrates that both the geometry of the container and environmental conditions significantly influence the evaporation rate of ethanolic cinnamon extract. The faster evaporation observed in larger surface area vessels (e.g., Petri plates and china dishes) aligns with basic physical principles, where increased surface area allows for greater exposure to air, facilitating faster solvent evaporation. In contrast, containers with smaller surface areas, such as test tubes, showed slower evaporation rates. Moreover, controlled environmental conditions, such as constant temperature and humidity, were found to reduce the evaporation rate, highlighting the importance of environmental control in preserving volatile compounds in herbal extracts.

The results also underscore the role of open conditions in accelerating evaporation, which could lead to the loss of volatile bioactive compounds. This finding has important implications for the storage and preservation of herbal extracts, where open-air exposure could compromise the integrity of the extract. These observations are consistent with previous studies that have shown how exposure to air and high temperatures can lead to the degradation of bioactive compounds in plant extracts.

4.2 Comparison with Existing Literature:

While previous research has explored the effects of environmental factors on the evaporation of herbal extracts, the focus on container geometry and surface area has been limited. Several studies have investigated the evaporation of essential oils, where container shape and surface area were found to influence evaporation rates, similar to the findings in this study. However, the impact of these factors on the phytochemical composition of ethanolic herbal extracts, particularly cinnamon, has not been extensively studied, making this research a novel contribution to the field.

Our findings align with the literature on the stability of bioactive compounds in plant extracts, where controlled conditions have been shown to better preserve these compounds compared to open exposure. The results from TLC analysis in this study further support the idea that environmental factors play a crucial role in preserving the phytochemical integrity of cinnamon extracts, particularly compounds such as cinnamaldehyde and eugenol, which are known for their bioactivity.

4.3 Phytochemical Stability and Bioactive Compound Retention:

The TLC analysis revealed a significant difference in the retention of bioactive compounds between the open and controlled conditions. Under open conditions, the intensity of the cinnamaldehyde and eugenol spots decreased over time, indicating that these compounds were likely evaporating or degrading. In contrast, under controlled conditions, the phytochemical composition remained more stable, suggesting that the preservation of bioactive compounds was better maintained when environmental factors such as temperature and humidity were regulated. This highlights the importance of controlling evaporation during the extraction and storage process to ensure that herbal extracts retain their therapeutic potential.

Fig. 09 Schematic representation of Comparision between all TLC plates

4.4 Practical Implications:

The findings of this study have direct implications for the formulation, storage, and preservation of cinnamon-based extracts, particularly in the pharmaceutical and food industries. The identification of container geometry as a critical factor in evaporation can help optimize the design of storage vessels for herbal extracts. Additionally, the role of controlled environmental conditions in preserving the bioactive compounds suggests that manufacturers of cinnamon-based products should consider these factors to maximize the efficacy of their products.

For example, pharmaceutical companies may consider using storage containers with smaller surface areas or employing airtight containers to reduce evaporation and preserve the therapeutic properties of cinnamon extracts. Similarly, the food industry could apply these findings to improve the shelf-life and quality of cinnamon-containing food products, ensuring that the beneficial compounds remain intact.

4.5 Limitations and Future Research:

While this study provides valuable insights into the evaporation dynamics and phytochemical stability of cinnamon extract, there are a few limitations. First, the study focused on ethanolic extracts, and the findings may not be directly applicable to other solvent extracts or essential oils, which may exhibit different evaporation behaviours. Additionally, the study did not examine the long-term stability of the extracts beyond 24 hours, which could provide further insights into the gradual degradation of phytochemicals over time.

Future research could expand on these findings by exploring the impact of different solvents, such as methanol or water, on the evaporation dynamics and stability of herbal extracts. Moreover, studies on the long-term storage of cinnamon extracts, including the effects of varying temperature and humidity over extended periods, would provide a more comprehensive understanding of the preservation techniques required for maintaining bioactive compound stability.

5. CONCLUSION:

This study provides valuable insights into the evaporation dynamics of ethanolic cinnamon extract and the impact of container geometry and environmental conditions on the preservation of bioactive compounds. The results demonstrated that both surface area and environmental factors play a critical role in the evaporation rate of herbal extracts, with larger surface areas promoting faster evaporation, particularly under open conditions. Additionally, the study showed that controlled environmental conditions effectively preserve the phytochemical integrity of cinnamon extracts, as evidenced by the more stable retention of bioactive compounds in TLC analysis. The findings have practical implications for the storage and preservation of herbal extracts, suggesting that the choice of container geometry and environmental conditions should be carefully considered to maintain the therapeutic efficacy of herbal products. This research contributes to the growing body of knowledge on the preservation of natural products and provides a foundation for optimizing the formulation, storage, and application of cinnamon-based extracts in the pharmaceutical, food, and cosmetic industries. Future research could expand on these findings by exploring the impact of different solvents, long-term storage conditions, and other herbal extracts, which would further enhance our understanding of the factors that influence the stability of bioactive compounds. Additionally, studies on the molecular mechanisms underlying the degradation of phytochemicals during evaporation could provide deeper insights into how to better preserve the therapeutic properties of herbal extracts.

6. COMPETING INTERESTS:

The authors declare that they have no competing interests.

7. CONTRIBUTIONS:

Katre SG is the only contributor behind the basic concept, literature survey, review, creation of manuscript content, examination, correction and editing of the manuscript; literature survey, writing the manuscript, referencing and citation. Chhangani JA contributed in review, corrections and editing of the manuscript. The author read and approved the final manuscript.

8. ACKNOWLEDGMENTS:

The authors are thankful to Mahatma Gandhi Institute for Rural Industrialization, Wardha and Ministry of MSME, Government of India for their constant support.

9. REFERENCES:

1. Proestos C. The Benefits of Plant Extracts for Human Health. Foods (Basel, Switzerland). 2020; 9(11). doi:10.3390/foods9111653

2. Oula Alzeina and Lina soubh. Evaluation of Polyphenols’ Inhibitory Activity of α-amylase in Cinnamon. Res J Sci Technol. 2019; 11(3): 174-178. doi:10.5958/2349-2988.2019.00026.3

3. Chetashri N. Patil, Swati U. Kolhe, Manthan R. Rode SSL andAsawari PM. Cinnamon an all-Inclusive Review: Detailed Examination of the Botanical Characteristics, Pharmacological properties, and Therapeutic Potential of Diverse Cinnamon Species. Asian J Res Pharm Sci. 2024; 14(3): 249-5. doi:10.52711/2231-5659.2024.00041

4. Purnendu Panda, Indu.S, Banamali Das KR. and MMR. Uses of Twak (Cinnamon) In Ayurveda with pharmacological evidence - A Review. Res J Pharmacol Pharmacodynamics. 2023; 15(3): 141-143. doi:10.52711/2321-5836.2023.00025

5. S. Sundar, K. Padmalatha, G. Helasri, M. Vasanthi, B. Sai Narmada, B. Lekhya RNJ and TS. Anti-microbial Activity of Aqueous Extract of Natural Preservatives- Cumin, Cinnamon, Coriander and Mint. Res J Pharm Technol. 2016; 9(7): 843-847. doi:10.5958/0974-360X.2016.00159.1

6. Sangavi. R GP and AK. Antibacterial Activity of Ethanolic extract of Cinnamon against clinical Isolates of Staphylococcus aureus. Res J Pharm Technol. 2019; 12(1):259-261. doi:10.5958/0974-360X.2019.00049.0

7. Neha Deokate and Rajendra Patil. Formulation and Evaluation of Antioxidant Cream of Cinnamon. Res J Top Cosmet Sci. 2024; 15(2): 75-78. doi:10.52711/2321-5844.2024.00013

8. Muhammad Hamza Ashfaq AS and SS. Antioxidant Activity of Cinnamon zeylanicum: (A Review). Asian J Pharm Res. 2021; 11(2): 106-6. doi:10.52711/2231-5691.2021.00021

9. Pathak R, Sharma H. A Review on Medicinal Uses of Cinnamomum verum (Cinnamon). J Drug Deliv Ther. 2021; 11: 161-166. doi:10.22270/jddt. v11i6-S.5145

10. Wang J, Su B, Jiang H, et al. Traditional uses, phytochemistry and pharmacological activities of the genus Cinnamomum (Lauraceae): A review. Fitoterapia. 2020; 146: 104675. doi: https://doi.org/10.1016/j.fitote.2020.104675

11. Almatroodi SA, Alsahli MA, Almatroudi A, et al. Cinnamon and its active compounds: A potential candidate in disease and tumour management through modulating various genes activity. Gene Reports. 2020; 21: 100966.

12. Ruchika Sharma, Neha Kumari M. A and CP. V. Standardization and Phytochemical Screening analysis for Herbal Extracts: Zingiber officinalis, Rosc., Curcuma longa Linn., Cinnamonum zeylanicum Nees., Piper longum, Linn., Boerhaavia diffussa Linn. Asian J Pharm Technol. 2020; 10(3): 127-133.

13. Mishra N, Srivastava R. Therapeutic and Pharmaceutical Potential of Cinnamon. In: 2020:124-136. doi:10.4018/978-1-7998-2524-1.ch010

14. Arampath PC, Dekker M. Thermal Effect, Diffusion, and Leaching of Health-Promoting Phytochemicals in Commercial Canning Process of Mango (Mangifera indica L.) and Pineapple (Ananas comosus L.). Foods (Basel, Switzerland). 2020;10(1). doi:10.3390/foods10010046

15. Liu Y, Jiang Q, Wang Q, et al. The divergence between potential and actual evapotranspiration: An insight from climate, water, and vegetation change. Sci Total Environ. 2022; 807: 150648. doi: https://doi.org/10.1016/j.scitotenv.2021.150648

16. Kazemi MA, Elliott JAW, Nobes DS. The influence of container geometry and thermal conductivity on evaporation of water at low pressures. Sci Rep. 2018; 8(1): 15121.

17. Ragya Eslavath, V. Harikrishna, N. Kosuru, Goli Venkateshwarlu MS and KK. Phytochemical Screening and TLC, UV-Spectrophotometer Study of Bougainvillea glabra. Asian J Pharm Anal. 2013; 3(3): 83-85.

18. Rucha A Patel, Meghna P. Patel HAR and NS. Forced Degradation Studies of Olmesartan Medoxomil and Characterization of Its Major Degradation Products by LC-MS/MS, NMR, IR and TLC. Asian J Pharm Anal. 2015; 5(3): 119-125.

Received on 15.07.2025 Revised on 19.08.2025

Accepted on 13.09.2025 Published on 08.10.2025

Available online from October 15, 2025

Asian Journal of Pharmaceutical Analysis. 2025; 15(4):256-262.

DOI: 10.52711/2231-5675.2025.00040

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

S. No.	Sample	Mobile Phase	Distance travelled by solute	Distance travelled by solvent	R_f value
1.	Standard ethanolic extract of cinnamon (at 0:00 hr)	Hexane: ethyl acetate (6:4)	3.6 cm	7.6 cm	0.47
2.		Hexane: ethyl acetate (7:3)	4.5 cm	7.6 cm	0.59
3.		Hexane: ethyl acetate (8:2)	5.1 cm	7.6 cm	0.67

Sr. No.	Apparatus	Distance travelled by substance	Distance travelled by solvent	R_f value
1.	Conical-flask	1.7 cm	7.6 cm	0.22
2.	Beaker	1.0 cm	7.6 cm	0.13
3.	Test-tube	1.3 cm	7.6 cm	0.17
4.	Petri-plate	0.5 cm	7.6 cm	0.065
5.	China-dish	0.7 cm	7.6 cm	0.092

S. No.	Apparatus	Distance travelled by substance	Distance travelled by solvent	R_f value
1.	Conical-flask	2.4 cm	7.6 cm	0.31
2.	Beaker	1.8 cm	7.6 cm	0.24
3.	Test-tube	2.1 cm	7.6 cm	0.27
4.	Petri-plate	1.4 cm	7.6 cm	0.18
5.	China-dish	1.8 cm	7.6 cm	0.24