Mariappan Gopalakrishnan*, Dhandapani Saravanan Antimicrobial Activity of ambonicus Herbal Finish on Cotton Fabric DOI: 10.5604/01.3001.0010.2854

Bannari Amman Institute of Technology, Abstract Sathyamangalam, Tamil Nadu, India-638401 An extract obtained from Coleus ambonicus was applied on cotton fabric by means of the * E-mail: [email protected] exhaust, micro encapsulation and nano encapsulation methods, and the antimicrobial activity of the finished fabric assessed quantitatively by the AATCC test method 100 against gram positive (Staphylococcus aureus) and gram negative (Escherichia coli) microbes. The fi- nish applied on the the samples using all three methods exhibit a good bacterial reduction percentage. The finish applied on the samples using all three methods possesses a higher bacterial reduction percentage against gram positive microbes than gram negative, even after washing. The method of washing conforms to ISO method 3. The wash durability of the antimicrobial activity was assessed by the bacterial reduction percentage after washing. The wash durability of the samples using the direct exhaust method was very poor and it lost its antimicrobial activity after 10 wash cycles. The wash durability of the samples using the micro encapsulated method shows antimicrobial activity up to 10 wash cycles, dropping gradually to very low levels at 20 wash cycles. The wash durability of the samples using nano encapsulation shows good antimicrobial activity against both gram positive and gram negative microbes even after 30 washes. Key words: Coleus ambonicus, cotton, micro encapsulation, nano encapsulation, antimi- crobial activity, bacterial reduction %, FTIR spectrum, wash durability.

release mechanism. The nano encapsula- Preparation of herbal Coleus tion method of finishing provides a high ambonicus extract surface area and extends the shelf-life of Collected Coleus ambonicus herbal the finish on cotton [11, 12]. leaves were shade dried to reduce the moisture content of the leaf to below Coleus ambonicus is a medicinal 10%. Unwanted materials like dirt and found abundantly in India. Six major other parts of were removed man- Introduction compounds account for 97% of the com- ually by hand. The dried herbal leafs position of the coleus ambonicus extract. Natural textile fibres like cotton are prone were crushed into small pieces and made Of these six, is the major com- to microbe growth. The moisture regain into powder, using a mixer. The ground pound and has antimicrobial, antifungal of cotton, the climate conditions in India herbal powder was then sieved using and insecticidal properties [13]. The oth- and the cellulose present in cotton favour fine mesh fabric to remove unground er flavonoid and phenol functional microbe growth [1, 2]. Microbial growth portions. 20 grams of the fine herbal groups present in the herbal extract may on textiles may cause foul smells, skin powder was refluxed in a Soxhlet ap- also contribute to antimicrobial activity irritation, cross infection, discolouration paratus (Borosil, India) in 200 ml of [14]. In this work, the antimicrobial ac- and colour stain. It also affects the func- methanol. The resultant supernatant was tivity of coleus ambonicus herbal finish tional properties of fabric like strength filtered with Whatman No.1 filter paper on cotton fabrics was analysed using the and elastic properties [3]. and the filtrate was maintained at room direct, micro encapsulation and nano en- temperature to remove organic solvents. capsulation methods. A wide range of synthetic antimicrobi- The fabric to be treated with the herbal

al agents like triclosan, metals and their extract was scoured with soap at 60 °C to remove any existing surface finish. salts, organometallics, phenols and qua- Materials and methods ternary ammonium compounds have The soap treated fabric was washed with been applied to cotton [4]. Synthetic Materials distilled water and air dried. The herbal antimicrobial agents have excellent an- In my study, 100% cotton fabric with extract prepared was applied to the cot- timicrobial efficacy for a wide range of a plain weave construction was used. ton fabric using the exhaust method with

microbes, but issues of eco-friendliness The fabric has 32 threads per centimeter a 1:30 material to liquor ratio at 60 °C for & biodegradability limit the use of syn- in the warp and weft directions. The yarn 30 minutes, after which the fabric was thetic antimicrobial agents [5]. Natural count used for this purpose was 15 tex in washed and dried in air. antimicrobial agents like neem, pome- the warp and weft directions. 115 grams granate fruit, tulsi leaf and etc., possess per square meter fabric density was used. Micro encapsulation of herbal extract very good resistance to most microbes The fabric was desized, scoured and Gum acacia was taken as wall material [6-10]. Most of the natural antimicrobial bleached with the standard recipe. for encapsulation of the Coleus amboni- agents are available in abundance and are cus extract. 10 grams of gum acacia was eco-friendly. Coleus ambonicus used as natural anti- allowed to swell for half an hour by mix- microbial agents was collected locally ing with 100 ml of hot water. 50 ml of The encapsulation process increases the from the Sathyamangalam village, Erode hot water was added to this mixture and life cycle of the finish by a controlled district, Tamil Nadu, India. stirred for 15 minutes at 40 °C to 50 °C

106 Gopalakrishnan M, Saravanan D. Antimicrobial Activity of Coleus ambonicus Herbal Finish on Cotton Fabric. FIBRES & TEXTILES in Eastern Europe 2017; 25, 4(124): 106-108. DOI: 10.5604/01.3001.0010.2854 temperature. 10 ml of core material Cole- a) b) us ambonicus was added to this mixture and stirred at 300-500 rpm for 15 min- utes. 10 ml of 20% sodium sulphate solu- tion was added slowly drop wise for 5-10 minutes. 5 ml of 17% formaldehyde was added at a slower stirrer speed. The stir- rer was then stopped and the mixture was freeze dried. The cotton fabric was im- mersed in the micro encapsulated herbal extract using the exhaust method with a 1:20 material to liquor ratio at 55 °C for 30 minutes and then dried at 80-85 °C in an oven. Figure 1. SEM photographs of fabric treated with nano encapsulated Coleus Ambon- Nano encapsulation of herbal extract icus at magnifications: a) 2000, b) 5000. The herbal extract prepared was encap- sulated with bovine serum albumin as the wall material by the coacervation process, followed by cross linking with glutaraldehyde. The Coleus ambonicus herbal extract was incubated for an hour at room temperature with the 2% bovine serum albumin protein powder required. A digital pH meter was used to adjust the pH to 5.5, by means of a 0.1M HCL. Ethanol was added slowly at 1 ml per minute into the solution to obtain a 2:1 ratio (v/v). The required amount of 25% glutaraldehyde was added and kept for two hours to allow the cross linking of protein. A rotary vaccum evaporator was used at reduced pressure to remove or- ganic solvents. The resultant nano parti- cles were purified by centrifuging at 4 °C and 10,000 rpm. The resulting pellets formed of nano capsules were suspend- ed in a phosphate buffer (pH: 7.4, 0.1M) and lyophilised with 2% w/v mannital. Figure 2. Particle size of nano encapsulated Coleus Ambonicus herbal powder. The nano capsules thus obtained were further dried by lyophilisation. The re- 37 + 2 °C for 24 hours. After incubation, assessment. A scanning electron micro- sultant nano encapsulated herbal powder the number of bacteria present in the scope was used to analyse the binding of was applied on cotton by the exhaust solution and percentage reduction (R) nano encapsulated particles on the fabric. method. Citric acid was used as a bind- were calculated using the following for- An FTIR8400S Spectrophotometer (Shi- er with a 1:20 material-to-liquor ratio at mula: madzu, Japan) was used to obtain FTIR 55 °C temperature for 30 minutes. Anti- R = 100(B-A)/B spectra to discover functional groups microbial activity of the finished fabric present in the sample. was assessed by the AATCC 100 test Where, A is the number of bacteria re- method. covered from inoculated treated test specimen swatches in a jar incubated Test methods over the contact period desired, and B Result and discussion Antimicrobial activity of the treated and the number of bacteria recovered from SEM analysis untreated samples was assessed quantita- inoculated treated test specimen swatch- Figure 1 shows SEM photographs of tively by the agar diffusion test method es in a jar immediately after inoculation nano encapsulated Coleus ambonicus at (AATCC 100). 4.8 + 0.1 cm circular cut (AATCC test method). The durability different magnifications. It is clear that treated, untreated and control samples of the finishing was assessed by analys- the nano encapsulated Coleus ambonicus were inoculated with the required quan- ing antimicrobial activity of the washed was attached to the fibre surface firmly. tity of AATCC specified gram positive samples. The washing of the sample was (Staphylococcus aureus) and gram neg- done in a laundrometer with the ISO 3 Figure 2 shows the diameter of the nano ative (Escherichia coli) test organisms. wash test method. After the specified encapsulated powder. The average diam- 100 + 1 ml of neutralising solution dis- number of washes, the test samples were eter of the nano encapsulated powder is tilled water was added and incubated at taken and washed thoroughly for further around 155.07 nm.

FIBRES & TEXTILES in Eastern Europe 2017, Vol. 25, 4(124) 107 Transmittance, % Transmittance,

Wavenumbers in, cm-1

Figure 3. FTIR Analysis of Coleus Ambonicus.

FTIR spectrum analysis quantitatively by the AATCC agar diffu- encapsulation methods possess a higher The peaks in the FTIR spectrum confirm sion test method 100, the results of which bacterial reduction percentage against functional groups present in the herb- are given in Table 1. The samples fin- gram positive microbes than gram neg- al powder. Figure 3 shows the FTIR ished with Coleus ambonicus in all three ative. This indicates that Coleus ambon- spectrum obtained from the Coleus am- methods show good antimicrobial activ- icus herbs are very active against gram bonicus herbal extract. The peaks 1580- ity. This may be due to the rich content positive rather than against gram nega- 1590 cm-1 substantiate the presence of of thymol in Coleus ambonicus herbs. tive. primary alkyl amide, 3600-3700 cm-1 Sivropoulou A et al. reported the antimi- – the existence of O-H phenolic com- crobial activity of Coleus ambonicus due Wash durability of finishing pounds, and 3000-2900 cm-1 revealed to the presence of thymol [13] and con- Table 2. show the wash durability of the presence of C-H in the aromatic phe- solacion. Y. Ragasa et al. reported three the samples finished by the direct,- mi nolic functional groups. Peaks at 2000- flavones: salvigenin, cirsimaritin and cro encapsulation and nano encapsu- 2200 cm-1 confirm the presence of flavo- chrysoeriol present in Coleus amboni- lation methods. A marked reduction in noids [15]. The presence of flavonoids cus herbs and that the presence of these the bacterial reduction percentage was provides the natural colour of the herbal flavones contributes to the antimicrobial observed in the sample finished by the extract and antimicrobial properties [16]. activity of the herbs [14]. direct exhaust method when increasing the wash cycles. The bacterial reduction The existence of these functional groups The samples finished by the direct- ex percentage was 15 and 12 for S. aureus in Coleus ambonicus is responsible for haust method show a better degree of and E. coli, respectively, after 10 wash the antimicrobial activity of the sample. bacterial reduction percentage for both cycles, confirming that the direct method gram positive (Staphylococcus aureus) of application has poor wash durability Antimicrobial activity and gram negative (Escherichia coli) of finishing. The bacterial reduction percentage of the than the micro encapsulation and nano sample finished using the direct exhaust encapsulation methods before washing. The micro and nano encapsulated sam- method, micro encapsulation method and The samples finished by the direct- ex ples show better resistivity in both gram nano encapsulation method was assessed haust, micro encapsulation and nano positive and gram negative microbes than for the direct method after washing. Table 1. Bacterial reduction percentage of the sample at zero wash cycles. It is evident that the nano encapsulated and micro encapsulated samples have re- Zero wash stricted release of anti microbial agents. S. aureus E. coli Direct method 100 100 The samples finished by the micro encap- Micro encapsulation 87 82 sulation method exhibit good resistance Nano encapsulation 85 81 to both gram positive and gram negative

Table 2. Bacterial reduction percentage of samples at different wash cycle.

Zero wash 5 wash 10 wash 20 wash 30 wash S. aureus E. coli S. aureus E. coli S. aureus E. coli S. aureus E. coli S. aureus E. coli Direct method 100 100 56 51 15 12 0 0 0 0 Micro encapsulation 87 82 78 73 65 58 18 11 0 0 Nano encapsulation 85 81 80 76 75 71 58 54 38 31

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