Due to the environmentally damaging effects of synthetic dyes and mordants, research has been carried out to introduce new methods and sources for the extraction of natural dyes and mordants. Three different flowers R. Indica, B. Glabra, and T. Erecta were selected to extract natural dyes. The Alo Vera gel and Azadirachta indica were used as natural mordants and compared with synthetic mordants namely Copper sulfate and Ferrous sulfate. The extraction was carried out by an incubator shaker and the optimum conditions obtained were 2 hours of shaking at 60℃. The antioxidant and antibacterial activity of liquid extracts was also performed. It was shown by the results that all the plants have the potential to show both activities. The TPC and TFC of all the plants were also noticeable. After the dye extraction, cotton fabric was dyed by them then the fabric was characterized by fastness to washing, and crocking. The results suggested that bougainvillea glabra is a better dye for dyeing purposes as compared to Rosa indica and Tagetes erecta. As a mordant Aloe Vera gel is proved to be better than Neem (Azadirachta indica).
Research Article
Extraction of chromophores from flowers of Rosa indica, Bougainvillea glabra, and Tagetes erecta and their application on cotton fabric
https://doi.org/10.21203/rs.3.rs-4822441/v1
This work is licensed under a CC BY 4.0 License
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Natural dyes
Rosa indica
Tagetes erecta
Bougainvillea glabra
mordants
In 1856, William Henry Perkin first introduced synthetic dyes, and at that time, synthetic dyes were widely accepted in the textile industry because of their low cost. Synthetic colorants are toxic or have a non-biodegradable nature. In the last few decades, synthetic dyes cause harmful effects on human health like carcinogenic and allergic reactions to the skin, to address the issue of toxicity the use of synthetic colorants is decreasing gradually. (Bhute, 2015). Synthetic dyes are organic compounds that are soluble in water, particularly those that are direct, acidic, basic, and reactive. The high solubility of synthetic dyes in water makes them difficult to remove by using traditional methods. The chromophoric group in the molecular structure of synthetic dyes can transmit the color to another substrate. The auxo chromic groups which are polar and can bind to the polar groups of textile fibers are responsible for the fixation of color to the material (Lellis et al., 2019). At present, a large number of synthetic dyes are used, around about 10,000,000 tons per annum. The manufacturing of synthetic dyes releases a large number of unfixed dyes. Moreover, synthetic dyes are not eco-friendly. Germany was the first country to impose a ban on the production and the use of specific azo dyes. The ban was also followed by some countries like India and Netherlands but synthetic pigments are still in use (Verma and Gupta, 2017).
The public’s growing awareness of environmental preservation, health concerns, and sustainability resulted in eco-friendly-resourced colorants that have created a new era in textile development and research. Furthermore, environmental maintenance concerns compelled western countries to use their advanced technical skills in textile materials for high-quality technical performance and production of cost-effective value-added textile products (Yusuf et al., 2017). The use of natural dyestuff instead of synthetic dyes is environmentally friendly and they do not pose any toxic effect on the wearer. The soft, soothing shades were produced by the use of natural dyestuff (Jordeva et al., 2020).
In the process of natural dyeing, there is one major problem natural dyes have limited sources for colorants but in the last few years, a lot of different methods were proceeded to find new roots for pigments of natural colorants from forestry and farming i.e., bark and woods, and also from food wastage like onion peels (Miah et al., 2017) and crushed barriers (Prabhu and Teli, 2014). Natural colorants are non-toxic, less pollutant, and have a less damaging effect on health. Natural colorants are renewable because they are obtained from natural sources and at the same time they are biodegradable. (Murugesh Babu and Ravindra, 2015). The shades of natural colorants are soothing to the human eye. Using different mordants or dyeing techniques with the same dye or with different concentration of mordants on the same dye gives us a variety of shades (Samanta et al., 2020).
In the dyeing process of textiles, natural colorants produce poor colors to improve the deficiency of color. Different types of mordant are used for textile dyeing. Metals ions are present in the mordant which play the role of an electron acceptor for the donor. Mordant forms a coordination bond with dye and also provides help in making a water-insoluble structure. The complex formation of mordant not only enhances the color of the fabric but also encourages the shades to maintain the colorants and make the colors look richer (Arora et al., 2017). Natural dyes are used in many fields like UV protective finishing, food colorants, beauty aids, sensitized solar cells, and insect-repellent finishing (Wangatia et al., 2015).
In this research work, the shaking method was used for the extraction of dyes from different flowers R. indica(red rose), T. erecta(marigold), and B. glabra(bougainvillea) dyed on cotton fabric using various mordants. Both natural and synthetic mordants were tested and compared for their efficiency against naturally extracted dyes.
Collection of samples
The flowers of Rosa indica (Rose), Bougainvillea glabra, Tagetes erecta (Marigold), Aloe vera, and leaves of Azadirachta indica were collected for extraction of mordants from local fields of Pakistan. All the collected samples, except Aloe vera, were washed and shade dried. Dried flowers and leaves were ground into a fine powder using an electronic grinding machine. For Aloe vera, the gel was separated mixed with water, and stored in an air-tight container in the refrigerator.
Extraction of colorants and mordant
Colorants and mordant were extracted from powdered samples of flowers and leaves by using the incubator shaker method. For this purpose, methanol was taken as a solvent with a Solid-liquid ratio (SLR) of 1:10 at 60℃ for 2 hours at 120rpm. After the process, the extract was filtered with Whatman filter paper no 42, and the extracts were used for further process to dye cotton fabric.
Assessment of Total Phenolic Content (TPC)
The TPC of methanolic extracts was evaluated by using the Folin–Ciocalteau’smethod.The concentrations of gallic acid were used to create the calibration curve. 1ml of aliquots of 0.01, 0.02, 0.03, 0.04, 0.06, 0.08, 0.09, and 0.10mg/ml solution of gallic acid in methanol were mixed with 5ml of Folin–Ciocalteau’s reagent (10 folds diluted) and 20% of sodium carbonate (4ml). After 1 hour the absorbance at 765nm was measured and the curve of the calibration was plotted by plotting the absorbance as the concentration functions. All the readings were taken in triplicate times.
Assessment of Total Flavonoids Content (TFC)
The TFC of extracted plants was evaluated by using the method described by (Rehman et al., 2013). Catechin was used as standard.0.5 ml of plant extract was incubated for 6 minutes with 2mL of distilled water and 0.15mL of 10% of NaNO2 solution and was added to the mixture and incubated for 6 minutes before 4% of the solution. The volume of the reaction mixture was increased to 5ml by adding methanol and mixing thoroughly. After 15 minutes of incubation, the absorbance of the reaction mixture was measured at 510 nm.
Antioxidant activity (DPPH)
Plant extracts antioxidant potential was assessed using the 2,2 Diphenyl 1-Picyrl hydrazine (DPPH) radical scavenging assay development. In this protocol 1ml of 0.004%, DPPH in methanol solution (freshly produced) was added to 3ml of plant extract and the combined solution was kept in dark for 30 minutes. The absorbance was then measured at 517nm.
High-Pressure Liquid Chromatography (HPLC)
For the detection of HPLC,a UV-Visible detector, 280nm is used at room temperature with the type of gradient HPLC in the range of 125mv, the column used was shim-pack CLC-ODS (C-18), 25cm×4.6mm, 5µm and the mobile phase on the column was a gradient ofA (H2O: acetic acid- 9.4:6, PH = 2.27) and B (Aceto nitrate 100%) for 0–15 minutes with the flow rate of 1mL/min(Oomen et al., 2020).
Fourier Transform Infrared Spectroscopy (FT-IR)
The fact that most molecules absorb light in the infrared region of the electromagnetic spectrum is the basis of FTIR. With the help of the interpretation of the spectrum of FTIR, it is possible to identify the functional groups and their respective chemical compounds in the extract. The presence of alkanes, ketones, aldehydes, alcohols, carboxylic acid, etc. was confirmed for dye extracts, column eluents, and dyed fabric (Shalini and Sampathkumar, 2012).
Determination of antimicrobial activity
Individual samples were tested against both bacterial strains, gram-negative Escherichia coli (E.coli ATCC 25922) and gram-positive Bacillus subtilus. The Institute of the Microbiology University of Agriculture Faisalabad, Pakistan confirmed the identity and purity of the samples. Bacterial strains were cultured in nutrient agar overnight at 37℃.The diffusion method was used to access the compound’s antibacterial activity. Concisely, 100L of tested micro-organisms suspension containing 107 colony forming units (CFU/ml) of bacteria cells nutrient agar medium individually impregnated with compound solution the filter disc (6mm) was placed on agar plates previously inoculated with the tested micro-organisms (Gesese et al., 2021).
Biological oxygen Demand
The amount of oxygen required by the micro-organisms for the bio-degradation of degradable conditions, carbonaceous organic matter present in the water through their bioprocess, biochemical and under the following reaction conditions five-day retention, darkness to avoid the presence of microscopic algae that produce oxygen by photosynthesis, consequently interfere with the results and temperature(Tara et al., 2019).
Chemical Oxygen Demand
COD was performed through the closed reflux method. A series of known standards are prepared using potassium hydrogen phthalate (KHP) for the completion of the chemical oxygen demand test because samples of wastewater will be in the high range standards of 100, 250, 500, and 1000mg/l are commonly prepared. COD heating block/ block are both activated to allow both instruments to stabilize. The COD test uses pre-prepared high-range (3-50ppm) or low-range (20-150ppm) vials based on expected results, if the expected results are unidentified, either range can be used(Deng et al., 2020).
Dyeing of Fabric
For dyeing scoured cotton fabric was provided by National Textile University. In this method, the fabric was pre-mordanted before dyeing. The mordant was taken and immersed in the water and then the fabric was soaked into the water the procedure was carried out for 1 hour at 90℃ using four mordants, two were natural, and two were synthetic mordants, and the results were compared. The fabric was then dyed by using three different extracts red rose, bougainvillea, and marigold. The procedure of dyeing was preceded for 1 hour at 80℃ on a dye bath with continuous stirring.
Statistical analysis
All spectroscopic analysis were performed in triplicates and one-way ANOVA (analysis of variance) was applied with α = 0.05 (Baljeet et al., 2016).
FTIR analysis of plant extracts
In the FTIR analysis of R. indica extract four different peaks were observed. The peak that was observed at 1014cm-1 shows the expected compounds of esters and amines. The peak that was observed at 1440cm-1indicates the alkanes group with CH2 bending (Fig. 1A). The observed peaks at 2838cm-1, and 2989cm-1 indicate the CH stretch with alkane. The weak peak that was observed at 1432cm-1 shows CH2 bending with the alkanes group. The medium peak at 1625cm-1 shows the ketone groups with carbonyl stretching while the other two sharp peaks at 2803cm-1, and 2983cm-1 indicate the presence of a carboxylic group that is acidic with OH stretch (Table 1). The observed peaks at 3089 in T.erecta indicate the CH stretch with alcohol (Fig. 1B).
| Sr# | Wavenumber cm− 1 (test sample) | Wavenumber cm− 1 (for reference) | Functional group assessment | Expected compound | ||
|---|---|---|---|---|---|---|
| 1. | R. indica | B. glabra | T. erecta | |||
| 2. | 1014 cm− 1 | ----- | ----- | 1300 − 1000 cm− 1 | C-O stretch and C-N stretch are often weak and hard to find | Esters, Amines |
| 3. | 1440 cm− 1 | 1432 cm− 1 | 1430 cm− 1 | 1600 − 1400 cm− 1 | C-H2 bending | Alkanes |
| 4. | ----- | 1625 cm− 1 | -------- | 1650 − 1600 cm− 1 | C = O stretch | Ketone |
| 5. | ----- | ----- | 1673 cm− 1 | 1680 − 1620 cm− 1 Often weak | C = C stretch | Alkenes |
| 6. | ----- | 2803 cm− 1 2983 cm− 1 | 2852 cm− 1 2964 cm− 1 | 3500 − 2400 cm− 1 | O-H stretch | Carboxylic acid |
| 7. | ----- | ----- | 3089 cm− 1 | 3500 − 3000 cm− 1 | C-H stretch | Alcohols |
| 8. | 2838 cm− 1 | ----- | ----- | 3000 − 2840 cm− 1 | C-H stretch | Alkane |
| 9. | 2989 cm− 1 | ----- | ----- | 3000 − 2840 cm− 1 | C-H stretch | Alkane |
The FTIR spectra analysis of B. glabra extract revealed five different peaks. The peak that was observed at 1430cm-1 shows the presence of alkanes with CH2 bending (Fig. 1C). the carboxylic group that is acidic with OH stretch, and the broad peak at 3089cm-1 indicates the compounds of alcohols with CH stretch (Nandiyanto et al., 2019).
Spectrophotometric analysis
Determination of TPC
The TPC in various extracts of flowers of T. erecta, R. indica, and B. glabra was estimated by Folin-Ciocalteau’s method using the standard as a Gallic acid. The calibration curve was built using the absorbance value obtained at several concentrations of Gallic acid. The regression equation of TPC was compute from equation (Y = 0.0055x + 0.0987 R2 = 0.9968) which was reported as mg gallic acid as equivalent (GAE) per gram of sample in dry weight (mg/g) (Fig. 2A & 2B). The R.indica contains a higher value of TPC as compared to T. erecta and B. glabra. The TPC value for the R. indica was 503.61 mg GAE/mL after the TPC value of B. glabra 404.05 mg GAE/mL was slightly higher than T. erecta 312.44 mg GAE/mL (Table 2).
| Sample | TPC mg GAE/mL | P-value (α = 0.05) | TFC in sample mg QE/g | P-value (α = 0.05) | % Inhibition | P-value (α = 0.05) |
|---|---|---|---|---|---|---|
| R. indica | 503.61 | P < 0.001 Significant | 78.20 | P < 0.001 Significant | 80.27 | P < 0.001 Significant |
| T. erecta | 312.44 | 58.41 | 49.69 | |||
| B.glabra | 401.72 | 95.1 | 60.49 |
The phenolic concentration of any plant is directly connected to antioxidant activity. The hydrogen donors or reducing agents and free-radical scavengers are all functions of phenolic compounds. The presence of a sizable amount of phenolic in the flower may have a substantial impact on the antioxidant qualities (Ghayempour et al., 2016).
Total flavonoid content
From the regression equation of the calibration curve (Y = 0.0038x + 0.0285), R2 = 0.9835 total flavonoids of the extract were computed which were expressed as quercetin equivalents (QE) per gram in the dry weight of the sample. The same trends will be shown in total flavonoid content as total phenolic content. The highest total flavonoids content value was shown by the flower B. glabra 95.10 mg QE/g and the lowest value 58.41 mg QE/g was given by the flower T. erecta (Table 2). The graphical data indicates that in comparison of the TPC, TFC and % inhibition the value of TPC was recorded highest while the lower value was noted in % inhibition (Fig. 3A & 3B).
The polarity of the extraction solvents has an impact on the concentration of flavonoids and phenol as well. Flavonoids are polyphenol compounds in plants that have structures of benzo-pyrone colorimetric with an aluminum chloride reagent that can be used to determine the total content of flavonoid extract (Jucá et al., 2020).
Antioxidant activity (DPPH)
The antioxidant activity of the different flowers was evaluated by DDPH free radical scavenging assay. In a concentration-dependent way, the radical scavenging capacity of several extracts increased. The highest antioxidant activity was shown by R. indica, at 81.27% and the lowest activity was given by T. erecta at 49.69%. The radical scavenging activity of different extracted flowers R. indica, T. erecta and B. glabra may be due to the presence of phenolic and flavonoid compounds and most of the activity is due to phenols. The damaging effect of oxidative stress is inhibited or avoided by the natural antioxidant potential of plants, among many others the DPPH assay (Nam et al., 2015) (Table 2).
HPLC analysis
The extracts of all the samples were analyzed both qualitatively and quantitatively via HPLC. The detection showed various compounds like quercetin, gallic acid, Caffeic acid, Vanillic acid, Chlorogenic acid, P-Coumaric acid, and Ferulic acid. In B. glabra and T. erecta quercetin, gallic acid, P-Coumaric acid, and many other compounds were commonly found. The amount of quercetin detected in R.indicais 78.31ppm, in B. glabra 67.8592ppm and T. erecta the amount is 528.0420 ppm and the amount of gallic acid is 45. 98ppm in R. indica, in B. glabra 88.6050 ppm and T. erecta, is 392.360 ppm. In T. erecta, the amount of quercetin and gallic acid is relatively higher as compared to R. indica and B. glabra (Table 3). Quercetin is a flavonoid and gallic acid is a subtype of aphenolic group containing antioxidant and coloring properties. Here, quercetin is a coloring compound and gallic acid is obtained from tannin responsible for the impartment of color(Gulzar et al., 2015).
| Compound name | Concentrations in ppm | ||
|---|---|---|---|
| R. indica | B. glabra | T. erecta | |
| Quercetin | 78.31ppm | 67.8592ppm | 528.0420ppm |
| Gallic acid | 45.98ppm | 88.6050ppm | 392.3260ppm |
| Caffeic acid | 35.87ppm | --- | --- |
| Vanillic acid | 44.59ppm | --- | 129.0367ppm |
| Chlorogenic acid | 71.57ppm | --- | 205.9679ppm |
| p-coumaric acid | 42.41ppm | 169.0388ppm | 28.9287ppm |
| Ferulic acid | 96.33ppm | 170.3029ppm | --- |
| Syringic acid | --- | 42.7299ppm | --- |
| Cinnamic acid | --- | 54.6295ppm | --- |
| Sinapic acid | --- | --- | 17.1618ppm |
| Benzoic acid | --- | --- | 219.9579ppm |
The fastness properties of Bougainvillea glabra dye are good as compared to the natural dye of Rosa indica, and Tagetes erecta however, the spectrophotometric analysis favors the natural dye of Rosa indica, and Tagetes erecta. In the case of fastness properties, natural dye of Bougainvillea glabra is considered good because it stays better on the fabric, the color doesn’t fade away and there is no excess of bleeding. And it is also mentioned above that the wet fastness of bougainvillea glabra extract was found good.
Analysis of color strength
Fastness properties
The fabric of cotton was dyed with the Pre-mordanting method and different grey scale rating was noted. Further, 4–5 staining of color on cotton fabric was observed on all dye samples. The shades obtained from the B. glabra flower as compared to other samples show maximum affinity towards the fabric of cotton. Analysis of natural dye B. glabra shows good washing fastness on a fabric by using Alovera gel as a mordant as compared to Neem. The crocking test of this dye shows dry fastness (4–5) better as compared to wet fastness (almost 3–4). The washing and crocking of the natural dye from T. erecta and R. indica flowers were not good. All the characterizations conclude that better results were shown with the natural dye of the B. glabra flower. The dye made from R. indica and T. erecta show poor color strength on the fabric. The shades obtained from the B. glabra flower show maximum affinity towards the fabric of cotton (Tables 4 & 5).
| Colorant source | Mordant | Washing fastness | Crocking | ||
|---|---|---|---|---|---|
| Stain of cotton | Shade of cotton | Dry | Wet | ||
| B. glabra | Azadirachta indica Aloe vera gel Copper sulfate ferrous sulfate | 4–5 4–5 4–5 4–5 | 2–3 4 1–2 1 | 3–4 4 3–4 3 | 4–5 4–5 3 4–5 |
| T. erecta | Azadirachta indica Aloe vera gel Copper sulfate ferrous sulfate | 2–3 4–5 2–3 4–5 | 1–2 1–2 1 1 | NIL | NIL |
| R. indica | Azadirachta indica Aloe vera gel Copper sulfate ferrous sulfate | 2–3 4–5 4–5 2–3 | 2 2–3 1 1 | NIL | NIL |
Table 5: Shows different shades obtained with different dyes and mordant on cotton fabric
In 2021 the researcher used the extraction method for the dyeing purpose with Rosa indica using different techniques of mordanting like post mordanting and pre-mordanting and make a comparison between them. Different analyses were also performed like fastness properties. The results of fastness properties are in accordance with previously described results where similar fastness rates were observed (Adeel et al., 2021).
The color properties of dyed cotton fabric based on CIELab color coordinates.
The dyed fabric were evaluated for their color coordinates and the results were represented in Table 5, 6, and 7 where L* is equal to light fastness a*is equal to red, green coordinates
| Dye | Mordants | L* | a* | b* | c* | h◦ | CV-SWL |
|---|---|---|---|---|---|---|---|
| R. indica | Azadirachta indica | 70.37 | 1.88 | 15.90 | 16.01 | 83.25 | 1.495 |
| Aloe vera gel | 82.38 | -0.58 | 17.61 | 17.62 | 91.88 | 0.922 | |
| Copper sulfate | 79.85 | -0.41 | 18.37 | 18.38 | 91.27 | 1.208 | |
| Ferrous sulfate | 63.75 | -4.61 | 17.92 | 18.50 | 104.43 | 2.766 | |
| T. erecta | Azadirachta indica | 82.14 | -0.60 | 17.85 | 17.86 | 91.94 | 0.932 |
| Aloe vera gel | 73.49 | 6.82 | 17.92 | 19.18 | 69.17 | 1.343 | |
| Copper sulfate | 70.85 | 1.67 | 14.82 | 14.92 | 72.93 | 10.309 | |
| Ferrous sulfate | 32.83 | 1.64 | 5.35 | 5.60 | 43.38 | 8.829 | |
| B. glabra | Azadirachta indica | 44.75 | 16.00 | 15.12 | 22.01 | 43.38 | 8.829 |
| Aloe vera gel | 47.01 | 14.55 | 15.90 | 21.55 | 47.53 | 7.878 | |
| Copper sulfate | 54.41 | 6.48 | 26.96 | 27.72 | 76.47 | 8.053 | |
| Ferrous sulfate | 55.94 | 5.91 | 25.39 | 26.07 | 76.89 | 6.748 |
| By | E. coli | Bacillus subtilus/Staphylococcus aureus |
|---|---|---|
| zone of inhibition in mm | ||
| R. indica | 21 | 19 |
| Positive control (ciprofloxacin) | 41 | 43 |
| T .erecta | 20 | 21 |
| Positive control (ciprofloxacin) | 39 | 44 |
| B. glabra | 3 | 5 |
| Positive control (ciprofloxacin) | 41 | 44 |
b* is equal to blue, yellow coordinates c* is equal to chromaticy h◦ is equal to hue angle and here CV-SWL is the color value at single wavelength in the terms of value, it is known as K/S(color strength). K/Scolor strength values were evaluated by spectrophometer. It measures different concentration for different dyed samples and their values will be compared. The dye extracted from R. indica produced lighter shades (L > 70) with Aloe vera gel, Azadirachta indica, and Copper sulfate mordant while Ferrous sulfate produced deeper shades with L (63.75). The net effect of its high Chroma value and lightness is reflected in its brilliant appearance.
Fabric dyed from extracted R. indica dye had a negative a*value with Aloe vera gel, Copper sulfate mordant, and Ferrous sulfate. While, CIE Lab color coordinates of T. erecta dye extract applied on cotton fabric using various mordants represents lighter shades (L > 70) with Aloe vera gel, Azadirachta indica, and Copper sulfate mordant while deeper shade was only produced with Ferrous sulfate L (32.83). Fabric dyed from extracted T. erecta dye only had a negative a*value with Ferrous sulfate. The dye extracted from B. glabra produced deeper shades (L > 70) with all mordants (Table 6).
In 2016, researcher performed an experiment on banana and banana appears to be the most promising in plentiful supply after citrus, the plant of banana was the most commonly used naturally produced product. During the research, there was an assumption that 41 percent of the outer part of the pseudo stem 3 percent of the peduncle, 9 percent of the floral stem, and 16 percent of the leaf and leaf stalk were present in plants of matured banana, according to the finding of the researcher after the harvest action of fruit nearly 89 percent of banana was wasted. The part of banana plants were also used as a mordant another natural dye for the coloration of cotton was pseudo stem sap and banana leaf. Plants of bananas were collected and crushed. Natural dye was extracted from the floral stems of the banana using a roller squeezer. The dyeing of the sample was carried out for different time duration like 60, 70, 80,90, and 100 minutes to investigate the temperature effect for the determination of variation of temperature on colorimetric appearances using CIE*L*a*b* color space different testing were performed lie color strength, the severity of color levelness, color coordinates and index of brightness rubbing fastness, light fastness Colorfastness, and washing fastness were used to evaluate the stability of dyed fabric bonding(Repon et al., 2016).
Antimicrobial Activity
In this research Natural dyes were tested against bacterial strains, including gram-negative Escherichia coli (E. coli ATCC 25922) and gram-positive Bacillus subtilus (B. subtilus). Bacterial strains were cultured in nutrient agar overnight at 37℃. The effect of dye concentration on antibacterial activity was further studied and, in each case, the zone of inhibition diameter was measured. Clear zones were seen for Bacillus subtilus. In Bacillus subtilus the positive control of B. glabra and T. erecta is 1% higher than the positive control in R. indica. But in E. coli the positive control of B. glabra than T. erecta and R. indica. The activity of T. erecta is much lower (Table 7). The antibacterial activity of sample extracts showed that they can be used in medical textiles and can provide benefits to society (Khanzada et al., 2020).
FTIR of dyed fabrics
Fabric dyed with B. glabra flower extract
FTIR analysis revealed a considerable difference in dyed fabric. However, these differences faded with complexes like synthetic mordants (copper sulfate, ferrous sulfate) and natural mordants (Neem, and Aloe vera gel) on cotton fabric. Variation in orientation and bond angle in the colored fabric may account for the difference in the reflectance curve.
Mordant ferrous sulfate
The broad peak at 3210.89cm− 1 could be OH and NH functional groups stretching frequently corresponding to carboxylic acids, alcohols, and amines. Another broad peak at 3162.98cm− 1, 2938.98cm− 1, and 2848.20cm− 1 with CH and OH stretch show the presence of alcohols and carboxylic group. The sharp peak at 1646.91cm− 1 indicates the expected compound of ketone with carbonyl stretching. The medium and weak peak at 1427.06cm− 1 and 1322.92cm− 1 indicates the presence of alkanes with CH2 bending and CO, CN stretch at esters and amines (Fig. 4A).
Mordant Aloe vera gel
The board peak at 2763.49cm− 1 shows the OH stretch with carboxylic groups and the weak peak at 1328.71cm− 1 shows the esters and amines group with CO and CN stretch (Fig. 4B).
Mordant copper sulfate
Four peaks were observed. The sharp peaks at 2624.64cm− 1 and 2359.58cm− 1 show the OH, stretch with the carboxylic acid group and carbon dioxide group. The medium peak at 1652.69cm− 1 indicates the group of the compounds of alkenes and the weak peak at 1334.49cm− 1 indicates the compound of esters and amines with CO and CN stretch (Fig. 5A).
Mordant Azadirachta indica leaves
The sharp peaks at 2794.34cm− 1 and 2582.21cm− 1 show the OH stretch with the carboxylic acid group. The narrow/ weak peak at 1319.07cm− 1 expects the phenol compounds with OH bending(Nandiyanto et al., 2019).
FT-IR and HPLC analysis extract of B. glabra dye showed that carboxylic acid and hydroxyl groups were present in it. However, these groups were also detected in the FT-IR analysis of dyed fabric. So, it was assumed that these functional groups present in the dye showed color to the fabric (Fig. 5B). The results of HPLC of B .glabra dye extract show the presence of quercetin contains hydroxyl group and in gallic acid carboxylic acid was also present as well as hydroxyl group. They were considered to be responsible for the impartment of color (Table 8).
| Sr# | Wavenumber cm− 1 (test sample) B.glabradye | Wavenumber cm− 1 (for reference) | Functional group assessment | Expected compound | |||
|---|---|---|---|---|---|---|---|
| Ferrous sulfate | Copper sulfate | Aloe vera gel | Azadirachta indicaleaves | ||||
| 1. | ----- | ----- | ----- | 1319.07cm− 1 | 1390 − 1310 cm− 1 | O-H bending | Phenol compounds |
| 2. | 1322.92 cm− 1 | 1334.49 cm− 1 | 1328.71cm− 1 | ----- | 1342 − 1266 cm− 1 | C-O stretch and C-N stretch are Often weak and hard to find) | Esters and amines |
| 3. | 1427.06 cm− 1 | ----- | ----- | ----- | 1600 − 1400 cm− 1 | C-H2 bending | Alkanes |
| 4. | 1646.91 cm− 1 | ----- | ----- | ----- | 1650 − 1600 cm− 1 | C = O stretch | ketone |
| 5. | ----- | 2359.58 cm− 1 | ----- | ----- | 2400 − 2000 cm− 1 | =C = O | carbon dioxide |
| 6. | ----- | 1652.69 cm− 1 | ----- | ----- | 1680 − 1620 cm− 1 | C = C stretch | alkenes |
| 7. | 2848.20 cm− 1 2938.98 cm− 1 | 2624.64 | 2763.49 cm− 1 | 2582.21 cm− 1 2794.34 cm− 1 | 3500 − 2400 cm− 1 | O-H stretch | Carboxylic acid |
| 8. | 3162.68 cm− 1 | ----- | ----- | ----- | 3500 − 3000 cm− 1 | C-H stretch | alcohols |
| 9. | 3210.89 cm− 1 | ----- | ----- | ----- | 3600 − 3200 cm− 1 | O-H stretch, N-H stretch | Alcohols, Carboxylic acid, and amines |
Biological oxygen demand
In this research work, BOD was performed to check the contamination of the natural dyes of B. glabra in wastewater using two different mordants one natural mordant Aloe vera gel and one synthetic mordant copper sulfate and the result of BOD was compared between these two mordants. The BOD value shown by the mordant Aloe vera gel is higher as compared to copper sulfate. Both contained contamination which means the quality was poor and the sample was polluted with a lot of organic matter. The level of BOD in mg/l 100 or more than 100 means water quality is much polluted. The BOD results indicate that both samples of natural dye but with different mordants contain contamination. The BOD value shown by the natural dye of the B. glabra flower with the mordant Aloe vera gel is higher as compared to synthetic mordant copper sulfate (Table 9).
| Sr # | Parameter | Unit | NEQS Guideline values | Biological oxygen demand |
|---|---|---|---|---|
| 1. | Aloe vera gel | mg/l | 80 | 1260 |
| 2. | Copper sulfate | mg/l | 80 | 1123 |
Chemical oxygen demand
Chemical oxygen demand will be performed in 24 hours to complete. COD was performed through the reflux method. In this research, COD was performed to check the contamination of the natural dyes in wastewater using two different mordants one natural mordant Aloe vera gel and one synthetic mordant copper sulfate with the same flower of B. glabra and the result of COD was compared between these two. The COD value shown by mordant Aloe vera gel is equal to synthetic copper sulfate mordant (Table 10).
| S. No | Parameter | unit | NEQS Guideline values | Chemical oxygen demand |
|---|---|---|---|---|
| 1. | Aloe vera gel | mg/l | 150 | ˃1500 |
| 2. | Copper sulfate | mg/l | 150 | ˃1500 |
In this study, the shaking method was used for extraction of dyes from different flowers and dyed on cotton fabric using various mordants. Both natural and synthetic mordants were tested and compared for their efficiency against naturally extracted dyes. In this research, the dyed fabric was tested using four mordants. Two fabrics were mordanted with the synthetic mordants Copper sulfate and Ferrous sulfate, and the other two were mordanted with natural mordants Neem (Azadirachta indica) and Aloe vera gel. The optimum temperature for dyeing of fabric was 80℃. The natural dye of R. indica and the flower of T. erecta showed poor color strength on the fabric.
The shades obtained from the natural dye of B. glabra flower extract showed maximum color strength on the fabric. Using different mordants for dyeing resulted in various kinds of shades. On comparing both natural mordants, it was observed that Aloe vera gel results were better than (Azadirachta indica). The R. indica only (Azadirachta indica) showed fair results to synthetic dyes. The flower of T. erecta and R. indica demonstrated poor results. Therefore, the crocking test for this flower was not performed because the washing results were not good (1–2).
Acknowledgement
The authors acknowledge Government College Women University, Faisalabad (GCWUF), Pakistan, and National Textile University, Faisalabad for their kind support.
Funding statement
All authors declare that there was no funding.
Competing Interest
All authors declare that there are no competing interests.
Consent for Publication
Not Applicable
Availability of data and materials
The datasets used and/or analyzed during the current study available from the corresponding author on reasonable request.
Authors' contributions
Faiza Nazir wrote the original manuscript perform the experiment and collected the plant samples; Ahsan Nazir supervised the study designed the experiments; Robass Ahmad analyzed worked on contents; Hafiza Affan Abid reviewed the literature, designed the graphs; Toqeer Abbas and Anis Ali Shah reviewed and edited the article; all authors revised the manuscript.
Voucher specimens
Plant specimens were identified by Dr. Iftikhar Ahmad associate professor of Botany, University of Sargodha, Sargodha, Pakistan and Dr. Mansoor Hameed, Professor at University of Agriculture Faisalabad. Voucher specimens are available at Institute of Plant Sciences, Department of Chemistry, Government College Women University Faisalabad, Pakistan, C0993-D40.
Statement regarding plant guidelines
Research regarding this manuscript was conducted inside the Department of Chemistry, Government College Women University Faisalabad, Pakistan the plants were treated by Faiza Nazir after the approval and guidelines from the departmental permission and standard methods were used to analyze the plant materials.
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No competing interests reported.