Plants serve as abundant sources of bioactive compounds, including polyphenols and carotenoids, with significant applications in the food, pharmaceutical and cosmetic industries. Various extraction techniques have been developed to efficiently isolate these valuable natural products for commercial use. Phytonutrients in fruits play a crucial role in mitigating the incidence of chronic diseases. Furthermore, home cooking often involves thermal or heat processing methods such as boiling, steaming, frying and freeze or microwave drying. The impact of these methods on the retention of bioactive compounds remains a subject of ongoing investigation. Some studies have documented a reduction in bioactive content following thermal treatment of fruits and vegetables. Heat processing such as steaming can enhance food digestibility, extractability and the bioavailability of essential nutrients by softening the food matrix. However, the overall effect of heat processing is significantly influenced by processing parameters and the inherent structure of the food matrix. Steaming is generally considered a safer method for preserving the bioavailability of vital nutrients in plant foods. A study by Mehmood et al. showed that antioxidant content in fruits and vegetables was higher in steaming compared to microwaving or boiling (Mehmood and Zeb, 2020).
Drying emerges as a preeminent method for preserving fruits, vegetables and herbs characterized by a significant reduction in volume and weight, consequently minimizing packaging, storage and transportation costs. Furthermore, this process induces modifications in flavour and texture, yielding a novel category of products such as snacks that can serve as healthier alternatives to commercially available confectionery. Notably, water removal effectively inhibits the proliferation of microorganisms and suppresses detrimental chemical reactions, thereby extending shelf life (Calín-Sánchez et al., 2020). Food dehydration, by definition, constituted a process involving the removal of water from food through the circulation of hot air. This process effectively deactivates enzymes and inhibits bacterial growth. Dried foods exhibit desirable attributes including enhanced taste, nutritional value, reduced weight and ease of preparation, storage and utilization. Drying exerts a minimal impact on the overall nutritional composition of food. While vitamin A is generally well-preserved during the drying process, its light sensitivity necessitates the storage of vitamin A-rich foods in dark environments. Yellow, orange and red-coloured fruits and vegetables such as Tamarillo are notable sources of β-carotene. Conversely, vitamin C is susceptible to degradation upon exposure to heat. However, pre-treating foods with citrus juices such as lemon, orange or pineapple can effectively enhance vitamin C retention. Dried fruits and vegetables, owing to their high fibre and carbohydrate content and low-fat content represent highly nutritious dietary choices. Freeze-drying also recognized as lyophilization or cryodesiccation, constitutes a dehydration technique commonly employed for preserving perishable materials and facilitating convenient transportation. This process involves freezing the material followed by a reduction in surrounding pressure, enabling the frozen water within the material to undergo sublimation, directly transitioning from the solid phase to the gaseous phase.
Tamarillo (Solanum betaceum), an underexploited fruit native to South America, is also cultivated in the Nilgiris District of Tamil Nadu, India. This egg-shaped, juicy fruit, characterized by a sour taste and containing numerous flat seeds, possesses a bitter peel. Commonly referred to as "tree tomato," it is utilized in the preparation of chutneys, sauces, pickles, jams and salads. The fruit is rich in essential nutrients, particularly fiber, vitamins and minerals and it contains significant levels of bioactive substances, including polyphenols and antioxidants, making it a valuable food for managing degenerative disorders. The phytonutrients and antioxidants present in the fruit exhibit anti-cancer properties by effectively scavenging free radicals. Regular consumption of Tamarillo fruit has been associated with reduced blood sugar levels, weight loss, prevention of kidney stones, and improved immunity. This research aims to investigate the impact of processing techniques used especially as freeze-drying and steam-cooking on the phytonutrient constituents and functional characters of Tamarillo. The research aimed to determine how these processing methods affect the presence and preservation of bioactive compounds including phenolic, flavonoids, glycosides and carotenoids that contribute to antioxidant and nutraceutical potential (Suganya and Kalpana, 2022). Fruit processing involves techniques such as freezing, drying, and cooking that help preserve fruits beyond their natural harvesting season. Tamarillo is a seasonal and underexploited fruit that is not always available fresh throughout the year. By applying processing methods like freeze-drying or cooking, the shelf life of the fruit can be extended, allowing it to be consumed even during off-seasons. This not only makes the fruit available year-round but also improves its usability in various forms such as powders, preserved foods, extracts, or functional food ingredients, thus increasing its application in food and nutraceutical industries. The present study was conducted to determine the effects of processing techniques on the functional properties and bioactive compounds present in Tamarillo.
Materials and Methods
Procurement of fruits
Ripe Tamarillo fruits were harvested from Gudalur in the Nilgiris district of Tamil Nadu. After collecting, the fruits were cleaned and stored at ambient temperature ranges from 20-25 ℃. They were then scaled, and cut into uniform pieces. Whole Tamarillo plant was authenticated by Botanical Survey of India, TamilNadu agriculture University, Coimbatore and the study obtained Institutional Human Ethical Clearance (AUW/IHE/FSN-22-23/XMT-8).
Sample preparation with freeze dried fruit
Foods can be preserved for extended periods through freezing or lyophilization, both of which effectively remove water from food samples. In this study, the freeze-drying technique was selected for drying the Tamarillo fruit samples. A total of 250 grams of fresh fruit, including the peel, was pre frozen for 12 hours and lyophilized at a temperature ranging from -45 °C to -50 °C for 42 hours under a vacuum pressure of 0.010 m to 0.012 m Torr. The resulting dried fruit samples were then ground into a powder kept in airtight container and stored in a frozen state for future use (Ahmed et al., 2013).
Sample preparation by steaming
Steaming involves cooking food over boiling water without the addition of fats (Vintilă, 2016). In this process, water was added to steamer and a perforated stand was placed inside the steamer. The water level was maintained below the stand to ensure effective steaming. The fruit was positioned on the perforated section of the stand, and the steamer was covered with a lid while cooking over medium-high heat. The fruit was cooked by the hot steam produced from the boiling water, with the samples prepared under atmospheric pressure. A total of 50 grams of fruit was steamed for 5 minutes at a temperature of 100 °C.
Identification of functional group using FTIR Spectroscopy
FTIR analysis was carried out using the extracted material at a wavelength of 3600 nm and a mid-infrared spectrum at a wavelength of 4000-600 nm. The functional group was determined using FTIR spectroscopy for the freeze-dried materials. One gram of the sample was collected, combined with 50 milliliters of 80% ethanol, stored in a shaking incubator and filtered through Whatman filter paper No. 40. The filtrate was collected in sterile bottles, centrifuged at 2500 rpm for 15 minutes and kept refrigerated at 5 °C until use (Sheybani et al., 2023).
Sample extraction with ultrasonic techniques
Five grams of processed samples were weighed and combined with 25 milliliters of concentrated methanol. The mixed materials were maintained in an ultrasonic homogenizer at 50–60 °C for 15-20 minutes after being vortexed for 10 minutes. Ultrasound waves as opposed to microwave (electromagnetic wave), are high-frequency sound waves that are audible to humans. An elastic medium such as liquid solvents, soft plant tissue, etc., is significantly impacted by sound waves. When sound waves pass through the medium, the shape of the medium changes and when sound waves are not there, the medium returns to its original shape. thus High frequency ultrasonic waves exert a piston-like effect on the medium (Panja, 2018). Cavitation bubbles are created inside the medium throughout the process; when they collapse, a significant amount of energy in these tiny bubbles are released, and localized hot and cold spots are produced. The cavitation effect is the mechanism used in phytochemical extraction.
Estimation of bioactive compounds
Phenolic compounds: The Folin-Ciocalteu method was used to determine the total phenolic content (TPC) in Tamarillo fruit samples. The process involved mixing 5 grams of the sample with 25 ml of 98% methanol, the mixture was then vortexed and ultrasonicated for 60 minutes. The extracts were filtered, centrifuged, and then 1 ml of the extracted sample solution was added to a Folin-Ciocalteu reagent. After 5 minutes at ambient temperature of 20-25 ℃, 1.5 ml of 20% sodium carbonate solution was added, and the mixture was stored in the dark for two hours at room temperature. A UV spectrophotometer assessed the sample absorbance at 765 nm against a control, and the results were extrapolated from a Gallic acid standard curve, expressed as milligrams of Gallic Acid Equivalent (GAE) per 100 grams of dry weight (Diep et al., 2020).
Flavonoids: A 10 ml of sample extracts were shaken vigorously with 4 ml of distilled water and allowed to keep for 5 minutes. Various aliquots of Quercetin working standard (100 g/ml in methanol) were prepared in separate test tubes, adjusting the total volume to 4 ml with distilled water including a blank sample as control. Then 0.3 mL of 5% sodium nitrate and 0.3 ml of 10% aluminum chloride were added to each tube, followed by shaking incubator at 37 °C for 10 minutes. Subsequently, 2 ml of 1 M sodium hydroxide and 10 ml of distilled water were added. The test tubes were shaken thoroughly, the mixture was filtered, and the absorbance of the reaction mixture was measured at 510 nm using a colorimeter against the blank. The total flavonoid content was quantified as milligrams of quercetin equivalent based on a standard curve (Rohilla and Mahanta, 2022).
Cardiac glycosides: A 10% of extract was combined with 10 ml of freshly prepared Baljet's reagent. After one hour, the mixture was diluted with 20 ml of distilled water, and absorbance was measured at 495 nm using a spectrophotometer. A standard curve was created using different concentrations of securidaside. The total glycoside content from triplicate measurements was expressed as milligrams of securidaside per gram of Tamarillo sample extracts (Dibulo et al., 2017).
Carotenoids: A sequential extraction was performed on a 5 g Tamarillo sample. This involved three successive extractions with 10 ml of hexane (containing 0.1% butylated hydroxytoluene, BHT) followed by three extractions with 10 ml of ethyl acetate (containing 0.1% BHT). After each extraction, the suspension was ultrasonicated for 15 minutes at moderate temperature in a dark environment and then centrifuged at 3000 rpm for 10-15 minutes. The resulting suspension was filtered through a 0.45-micrometer syringe filter. The collected filtrate was evaporated to dryness under vacuum and re-dissolved in methanol. The absorption of the mixture was measured at 425 nm, and the total carotenoids content was calculated using the standard curve. The results are expressed in terms of β-carotene as µg per 100 g of the Tamarillo sample (Giuffrida et al., 2018).
Data analysis
Paired sample t-test was applied to identify the statistical difference between the effects of drying and cooking on the bioactive compounds of tamarillo fruits. Sigmaplot 14.5 software was used to analyze the statistical values.
Results
Identification of functional groups using FTIR spectroscopy
Functional groups were identified by FTIR Spectroscopy and the results were presented in Table 1 and Figures 1 and 2.
Freeze-dried and steam cooked Tamarillo fruit samples had more or less unique spectroscopic signatures. The FTIR spectra of the following functional groups can be used to confirm
the presence of phenolic substances in the
extracts of Tamarillo fruit samples: OH alcoholic/phenolic (3400-3200 cm-1), C=C aromatic (1650-1600cm-1), C-H aromatic (700-420cm-1). The peak values and their functional groups are depicted in Table 1 and Figures 1 and 2. Analysis of freeze-dried and steam-cooked Tamarillo fruit samples reveals distinct spectroscopic signatures, with FTIR Spectroscopy spectra, confirming the presence of phenolic substances in Tamarillo fruit samples. Key functional groups identified include OH alcoholic/phenolic groups (3400-3200 cm-1), C=C aromatic groups (1650-1600 cm-1), and C-H aromatic groups (700-420 cm-1). The FTIR spectra of freeze-dried and steam-cooked Tamarillo fruit samples exhibit similar bands but with varying magnitudes, reflecting differences in composition.
| Table 1. FTIR peak values and functional groupings of Tamarillo. |
|
| Peak values (cm-1) |
Functional group |
| 3695.61 |
Phenols |
| 2970.38 |
Alkenes |
| 1743.65 |
Aliphatic acid / polysaccharides |
| 1450.47 |
Aromatic / polysacahraides |
| 1381.03 |
Nitro compound / cutin / waxes |
| 1273.02 |
Ether / polysaccharides and phenolics |
| 1049.28 |
Esters / polysaccharides |
| 879.54 |
Carboxylic acids |