Eco-Dyeing and Functional Finishing of Cotton Fabric Using a Natural Colour Derived From Lotus Seed: Enhanced Fastness Properties with Chitosan

Medicinal plant-based dyeing introduces vibrant colours alongside antibacterial and anti-odor properties, fostering sustainability. Innovations include mass sampling for consistent pigment production. Furthermore, lotus seedpods, rich in bioactive compounds, exhibit potential in diverse fields. Chitosan, a biopolymer derived from chitin, boasts versatility in drug delivery systems, wound healing, and antimicrobial applications. This experimental inquiry delves into the dyeability and antimicrobial efficacy of cotton fabric subjected to treatment with chitosan and dyed using Lotus seeds. The investigation encompasses varied parameters such as dye concentration, duration, and temperature for fabric dyeing. The study elucidates the aqueous extraction methodology for dye from Lotus seeds, the chitosan-based mordanting techniques. Standardized assessments are utilized to gauge colour fastness characteristics, encompassing washing, rubbing, and perspiration fastness, along with assessing the antimicrobial attributes of the treated cotton fabric. The study indicates that the application of chitosan-treated cotton fabric enhances dye intensity and overall dyeing quality, consequently improving the colour fastness property of washing, rubbing, and perspiration. Additionally, the treated cotton fabric exhibits significant antimicrobial activity against common bacteria, highlighting the potential of Lotus seed-derived colourants and chitosan mordant in the development of sustainable and antimicrobial textiles. The future scope lies in optimizing the integration of Lotus seed-derived colourants and chitosan mordant for enhanced sustainability and antimicrobial efficacy in textile applications, leveraging advancements in dyeing technology and antimicrobial formulation techniques.


INTRODUCTION
Natural dyeing has a lengthy history, having been utilized since ancient times.Initially, natural dyes were obtained from plants, insects, and animals.The initial use of natural dyes on textile fibres is estimated to have begun in Mesopotamia and India around 4000 BC.However, with the discovery of synthetic dyes in the 19 th century, the use of natural dyes decreased significantly.Synthetic dyes offered a wider range of colours and were easier to use.Additionally, industrialization and https://doi.org/10.31881/TLR.2024.099globalization led to an increase in the use of synthetic dyes [1].However, in recent years, there has been a resurgence of interest in natural dyes due to growing concerns about the environment and health hazards associated with synthetic dyes.Natural dyes are considered eco-friendly and more sustainable, and there is a niche market for textiles coloured with natural dyes.Natural dyeing is the process of dyeing textile fibres and fabrics using plant materials.Natural dyes are derived from various plant components such as leaves, bark, flowers, fruits, and roots [2].They possess unique qualities such as calming colours, biodegradability, safety, and antimicrobial resilience.However, the usage of natural dyes has decreased due to the availability of inexpensive synthetic dyes.Nevertheless, there is a growing interest in reviving the use of natural dyes, especially in the field of textile arts [3].Recent research focuses on the application of natural dyes in dyeing textile fabrics to reduce health hazards and pollution, as well as enhance dyeing quality and antimicrobial activity.Challenges in natural dyeing include difficulty in colour reproduction and the need for education about natural colourant use.
Innovations in natural dye production include mass sampling of dye plants for consistent pigment production [4].
Natural dyeing with medicinal plants is a sustainable and eco-friendly method that offers not only vibrant colours but also medicinal properties.Various plants such as Hemigraphis colourata, Bacopa monnieri, Lawsonia inermis, Hibiscus sabdariffa, Rheum officinale, Sambucus nigra, and Chamomilla recutita have been studied for their dyeing potential and medicinal characteristics.These plants have been used to develop herbal composites and dye solutions that can be applied to fabric samples using different techniques such as pad-dry cure process, radiation methods, and ultrasonic dyeing equipment [5].The finished fabrics show good antibacterial activity against Escherichia coli and Staphylococcus aureus, as well as anti-odour properties.The natural dyeing method using plant materials has a lower allergenic potential compared to synthetic methods.However, achieving full coverage of hair colour with plant materials may be challenging, and the selection of additives with appropriate pH values is important for favourable results [6].
Natural dyeing involves the use of plant extracts and compounds to colour textiles.Different natural dyes have been studied, and their active compounds have been identified.For example, madder root extract contains salicylic acid, quercetin, ellagic acid, and benzoic acid as main compounds [7][8].
Cochineal extract contains rutin, kampherol, myricetin, quercetin, and salicylic acid as main compounds.Impatiens balsamina extract, chloroacetic acid, and an organic carboxylic acid compound are used in a natural plant textile fabric dyeing agent [9].A natural textile dye includes a garden balsam extract, Cinnamomum camphora leaf extraction liquid, and an organic carboxylic acid compound.
These compounds contribute to the colour and properties of the dyed fabrics, providing stability, brightness, and environmental friendliness [10].https://doi.org/10.31881/TLR.2024.099 Lotus seedpods are the by-products of lotus plants and have been studied for their phytochemicals, biological activities, and industrial applications.They contain various phytochemicals such as proanthocyanidins, flavonoids, alkaloids, and terpenoids, which exhibit bioactivities like antioxidation, antibacterial, and neuroprotection.Additionally, lotus seedpods have been explored for their potential as a source of polysaccharides, which have shown antioxidant and α-glucosidase inhibitory effects, making them suitable for use as natural antioxidants and hypoglycemic substitutes.Furthermore, gasassisted combined with glycerol extraction (GAGE) has been proposed as a green and efficient method for recovering polyphenols from lotus seedpods, which have demonstrated antioxidant activities [11].
Overall, lotus seedpods have shown promise in various fields such as the food industry, medicine, and environmental sustainability.Lotus seedpods have various medicinal uses.They are rich in phytochemicals such as proanthocyanidins, flavonoids, alkaloids, and terpenoids, which exhibit bioactivities like ameliorating cognitive impairment, antioxidation, antibacterial, anti-glycative, neuroprotection, and anti-tyrosinase activities [12].The seed epicarp of Nelumbo nucifera Gaertn.has been found to have higher extractable total phenolic content and flavonoid content compared to the seed and seed pod.It also showed higher antioxidant activity, including DPPH and ABTS radical scavenging, reducing power, and hydrogen peroxide scavenging activities [11].Lotus seedpod extract has been shown to have hepatoprotective effects, reducing intracellular lipid accumulation, oxidative stress, and apoptosis in human hepatocytes.Additionally, lotus seedpod extract has been used for the treatment and prevention of kidney diseases, as it can suppress inflammatory factors, enhance antioxidant enzyme activity, and reduce kidney oxidative pressure [13][14].Overall, lotus seedpods are considered a functional food with multiple therapeutic benefits, including anti-adipogenic, antioxidant, antitumor, cardiovascular, hepato-protective, anti-inflammatory, and hypoglycemic effects.
Chitosan is a biopolymer derived from chitin, and it has gained attention in various fields due to its unique properties.It is biocompatible, biodegradable, and can be easily modified for specific applications.Chitosan has been extensively used in drug delivery systems, allowing for targeted and sustained release of drugs.Chemical modifications have improved its solubility, stability, and functionalization ability, making it suitable for biomedical applications [15,16].Chitosan's versatile properties, such as antibacterial, antifungal, and non-toxicity, make it a smart material for different applications, including wound healing, tissue engineering, and gene therapy [17].https://doi.org/10.31881/TLR.2024.099Its ability to encapsulate and deliver substances has led to advancements in medical and pharmaceutical applications.Chitosan also finds applications in the textile industry, where it imparts antimicrobial and other biological activities to fibres and fabrics [18].Chitosan, a naturally occurring polymer derived from chitin found in the shells of animals, has various medicinal uses.It has been studied for its non-toxicity, biodegradability, biocompatibility, immunostimulant, anticancer, antibacterial, and antimicrobial activity [19].Chitosan can be used in the treatment and prevention of various illnesses, as well as in diagnostic and therapeutic applications for managing malignancy.
Chitosan can also be combined with other active substances, such as metals, drugs, and natural compounds, to enhance its antibacterial effects and application potential [20].Additionally, chitosan derivatives, such as chitosan Schiff bases, have shown increased antimicrobial activity against bacteria and yeasts, making them potential candidates for antimicrobial prevention in various fields [21].
Chitosan and its derivatives have also been extensively studied for their antibacterial, antitumor, https://doi.org/10.31881/TLR.2024.099antioxidant, and tissue regeneration effects, making them suitable for drug delivery and wound healing applications.Overall, chitosan and its derivatives have promising potential in the medical field for various therapeutic effects and drug delivery functions [22][23][24].Today, the global emphasis on natural dyeing and sustainability is growing exponentially.Increasing awareness among people has underscored the necessity for textiles dyed with natural substances.In a recent study, I explored the potential of lotus seeds as a natural dye for cotton fabric.To enhance colour retention and durability, the cotton fabric was meta-treated with Chitosan powder.
Subsequently, the treated samples underwent rigorous testing to evaluate various properties such as washing, rubbing, and lightfastness.Additionally, antimicrobial properties were assessed to gauge the fabric's potential to resist microbial growth.https://doi.org/10.31881/TLR.2024.099

Materials and Methods
In this experimental study, exclusively 100% cotton fabric served as the primary material.The procurement of the fabric took place at the local market situated in Pune, Maharashtra.Additionally, the essential components for the experiment, namely lotus seed powder and Chitosan, were also sourced from Pune's local market.

Aqueous extraction
The extraction of dye from lotus seeds was carried out using the aqueous method.Initially, a beaker containing 300ml of water was prepared.Once the dyeing bath temperature reached 50 degrees Celsius, the lotus seed powder was introduced.Fifteen grams of dye were incorporated into the 300ml water.The dye solution underwent boiling at 70 o C for 40 minutes.Subsequently, the solution was subjected to double filtration through filter paper, resulting in a prepared and refined dye solution suitable for the dyeing process.

Mordanting and dyeing Procedure
The experiment involved treating samples under various time and temperature conditions, employing chitosan as a mordanting agent.Chitosan, a weak base, exhibits insolubility in water and organic solvents but can be dissolved in dilute aqueous acidic solutions (pH < 6.5), facilitating the conversion of glucosamine units into a soluble form, R-NH3+.Meta mordanting was conducted before dyeing the samples, aiming to enhance the dye's fixation onto the material.The dyeing process relied on three primary factors: dye concentration, time, and temperature.Dye concentration levels were categorized as low (15 g), medium (20 g), and high (25 g), while time durations were classified as short (70 min), medium (80 min), and long (90 min), and temperatures were categorized as low (60 °C), medium (70 °C), and high (80 °C) [25,26].

Colour fastness assessment
The study examined the varied colour fastness properties of woven cotton fabrics.The aspect of washability, which is a crucial element in textile appraisal, refers to the ability of a dyed or printed fabric to retain its colour fastness after undergoing laundering processes.Standardized protocols, such as ISO 105-C06:2010, are playing a pivotal role in the execution of systematic washing fastness tests, thus, they are making the test more reliable and comparable.Another very important element in the textile evaluation is rubbing fastness, especially in the context of dyed or printed textiles, as it is a measure of the ability of the fabric colour to resist rubbing or abrasion, which mimics real-world wear and laundering conditions.The approved AATCC Test Method 8-2016 by the American Association of Textile Chemists and Colourists (AATCC) defines processes for measuring the colourfastness to rubbing using a crockmeter device under both dry and wet conditions.Similarly, standardized procedures are available for assessing perspiration fastness, and are developed to achieve consistent and reliable results.One widely acknowledged standard in this domain is the ISO 105-E04:2013, which was adopted by the International Organization for Standardization (ISO) and is used to evaluate the colourfastness of textiles in acidic and alkaline perspiration.

Antimicrobial Test
AATCC Test Method 100-2019, a guideline from the American Association of Textile Chemists and Colourists (AATCC), defines an approach which quantifies reductions in bacterial populations due to exposure to treated materials [27].The broad process described in 'AATCC Test procedure 100-1999, Antibacterial Finishes on Textile Materials: Assessment of' with changes served as the foundation for the technique used to assess the qualitative and quantitative antibacterial properties of textile materials.As indicated in the AATCC Test Method, textile swatches were 'inoculated' with a test or challenge bacterium.After incubation, the bacteria were eluted from the swatches with defined https://doi.org/10.31881/TLR.2024.099quantities of extraction solution.The amount of bacteria in this extraction solution was then quantified for each treated textile sample to allow for comparison.In the AATCC Test Method, the treated specimen's per cent decrease was determined and compared.The test approach was discovered to be more versatile and When just the final population densities were compared, the process performed successfully [28].

Washing Fastness
The experiment that we performed involved the dyeing of the samples with lotus seed dye.Then, we compared whether they absorbed the dye better after the previous mordanting or not using chitosan.
Mordanting was the process of treating the fabric with a mordant, which was the substance that enhanced the dye-fibre association, hence resulting in dye-fastness and intense colour.https://doi.org/10.31881/TLR.2024.099 In the case of non-mordanted samples, the grades ranged between 2 to 3 respectively, and mordanted samples had grades ranging from 4 to 5.These differences indicated that mordanting with chitosan led to improved dye intensity and overall dyeing quality of the textile material when compared to nonmordanted fabrics.
The findings showed that mordanting with chitosan was a potential way for the dyeing process and produced great fastness properties.This upgrade was more evident in samples with higher dye concentrations, longer dyeing times, and elevated temperatures than left samples, which always achieved higher fastness grades compared to their un-mordanted counterparts.This implied that chitosan mordanting functionalized the lotus seed dyeing, which increased the adherence to the fabric and consequently irradiated more durable.

Rubbing fastnesses
The rubbing fastness properties of cotton fabric dyed with a natural colour derived from Lotus seed and enhanced dyeability with chitosan were assessed through a comprehensive series of tests.Nine fabric samples denoted as S1 to S9, underwent varying dye concentrations, time durations, and temperatures to evaluate their rubbing fastness performance.In the wet rubbing test without mordanting, the results ranged from grade 2 to 3, indicating moderate to good resistance to rubbing.
However, upon mordanting, there was a notable improvement in wet rubbing fastness, with most samples achieving grades 4 to 5, indicating excellent resistance to wet rubbing.
Similarly, in the dry rubbing test without mordanting, the samples showed fair to good rubbing fastness, ranging from grade 2 to 3. Yet, after mordanting, the dry rubbing fastness significantly improved, with most samples achieving grades 4 to 5, signifying excellent resistance to dry rubbing.It 0 is worth noting that the highest dye concentration (25 g) generally resulted in superior rubbing fastness properties compared to lower concentrations (15 g and 20 g), particularly when mordanted.
Additionally, longer dyeing times and higher temperatures tended to enhance rubbing fastness, as evidenced by the higher grades achieved in samples subjected to prolonged dyeing times and elevated temperatures.
These findings underscore the efficacy of Lotus seed-derived natural colour, combined with chitosan as a dyeability enhancer, in producing cotton fabrics with exceptional rubbing fastness properties.This is especially evident when optimal dye concentrations, dyeing times, and temperatures are employed, alongside mordanting treatments.

Perspiration fastness
The perspiration fastness of cotton fabric dyed with a natural colour derived from Lotus seeds and enhanced dyeability with chitosan mordant was evaluated using a grading system ranging from 1 (very poor) to 5 (excellent).The results, as depicted in the provided table, illustrate the varying grades achieved under different conditions.Across the samples, the application of chitosan mordant generally resulted in improved perspiration fastness compared to un-mordanted samples.Specifically, samples treated with chitosan displayed grades ranging from fair to excellent, with most achieving grades of 4 to 5.This suggests a positive impact of chitosan mordant in enhancing the perspiration fastness of the Lotus seed-derived natural colour on cotton fabric.
Furthermore, variations in dye concentration, time, and temperature during the dyeing process influenced the resulting colourfastness grades.Higher concentrations of dye and longer dyeing times tended to yield better perspiration fastness grades, particularly when combined with chitosan mordant treatment.Additionally, higher dyeing temperatures generally correlated with improved colour retention under perspiration conditions.
These findings underscore the potential of Lotus seed-derived natural colour, augmented with chitosan mordant, as a promising option for dyeing cotton fabric with satisfactory perspiration fastness, contributing to the development of more durable and sustainable textile materials.

Antimicrobial properties
In this study, the antimicrobial efficacy of cotton fabric treated with a natural colour derived from Lotus seed, enhanced with chitosan mordant, was investigated.The experiment aimed to assess the dyeability of the fabric as well as its antimicrobial properties following treatment.The cotton fabric underwent a dyeing process using the natural colour extracted from Lotus seed, and chitosan was employed as a mordant to enhance the dyeing process and potentially impart additional antimicrobial properties.
The results of the antimicrobial tests revealed promising outcomes.The treated cotton fabric exhibited notable antimicrobial activity against a range of microorganisms.This antimicrobial effect is attributed to the inherent properties of Lotus seed-derived colourant and the supplementary enhancement provided by chitosan as a mordant.The combination of these components not only facilitated effective dyeing of the cotton fabric but also conferred antimicrobial functionality, thereby potentially expanding the utility of cotton textiles in various applications where microbial control is desired.applications of such treated textiles in diverse fields, including healthcare, apparel, and home textiles, where antimicrobial functionality is of paramount importance.

Field Emission Scanning Electron Microscopy
Field Emission Scanning Electron Microscopy is an advanced imaging method utilized for examining the surface morphology and structure of materials with exceptional precision.By employing a stream of electrons, it generates high-resolution images, offering intricate details regarding the sample's surface topography.Analysis of SEM images has revealed the remarkable antimicrobial efficacy of Lotus seeds, along with the consistent application of uniformity onto the fabric surface.

Figure 1 .
Figure 1.Mechanism Between Natural Dyes and Fabric

Figure 7 .
Figure 7. Antimicrobial results against Staphylococcus aureus and Klebsiella pneumonia Figures 7 to 10 exhibit compelling evidence, clearly showcasing the contrast between surfaces treated with and without the mordant.

Table 1 .
Research Studies on Lotus Methods and Results

Table 2 .
Research Studies on Chitosan Methods and Results

Table 4 .
Sample coding for dyeing the samples