Indian Journal of Natural Products and Resources

Vol. 5 (4), December 2014, pp. 332-337

Mass transfer enhancement through optimized extraction of a natural dye from

Bougainvillea glabra Juss. bracts
P D Patil1*, C R Rao2, A I Wasif3, S V Anekar4 and J R Nagla3
1
Department of Technology, Shivaji University, Kolhapur, Maharashtra, India
2
Vigyan University, Vadlamudi, Guntur, Andhra Pradesh, India
3
Textile and Engineering Institute, DKTES, Ichalkaranji, Maharashtra
4
TKIET Warananagar, Maharashtra
Received 28 March 2014; Accepted 30 August 2014

Synthetic dyes are today’s established dyes for applications such as textile coloration. However, there are some serious
drawbacks of these new dyes. Therefore, there is increasing interest in the natural dyes. The present study was emphasized to use
the optimized extraction conditions so as to get maximum amount of the dye from bracts of Bougainvillea glabra Juss. Taguchi
design was implemented to know optimum extraction conditions: 150 micron particle size, 1:20 solid-solvent ratio, one hour
extraction and enzyme addition as assistance for extraction. The dye was extracted from both optimized and control experiments.
Fourier Transform Infrared Spectroscopy and UV-VIS spectroscopy techniques were used to identify chemical nature of the
extracted dye. The absorbance, mass transfer rates and dye yields were compared. The optimized extraction showed mass transfer
rate of about 3.3 times enhanced than that in the control extraction. The yield was 9.88 % more in case of optimized extraction.

Keywords: Bougainvillea glabra, Natural Dye, Taguchi optimization, Mass transfer enhancement.
IPC code; Int. cl. (2014.01)−A61K 36/00

Introduction Ever increasing environment related consciousness has

Natural dyes are the colorants extracted from been making natural dyes more popular like other natural
vegetative matter, animal residues or from minerals. products. Most of the natural dyes are biodegradable,
The dyestuffs for these colorants include roots, leaves, non-toxic and eco-friendly in nature. The other benefit of
barks, trunks, fruits and flowers of plants, animal introducing natural dyes is the significant reduction in the
sources like cochineal and shellfish or sometimes amount of toxic effluent resulting from the synthetic dye
from soils or clay. These dyes were used in the past processes7-8. At present, the international consumption of
for various purposes such as coloration of clothing, natural dyes is hardly 1% of the synthetic dyes. Prime
food, leather tanning and painting1-3. However, in the limitations and technical drawbacks of the natural dyes
mid 1800’s, with the invention of synthetic aniline, include low color yield and inefficient dye extraction
alizarin and indigo, natural dyes lost their economic methods. It has been the major problem to reintroduce
significance. Today, synthetic dyes have dominance these old dyes9. Therefore, the greatest challenge before
because of their varied colors, vast and easy us is to increase the yield of these dyes. Present study has
production and very good fastness properties. been carried out to address this particular issue. The plant
Advantages of the synthetic dyes compelled us to part selected for study is bracts of the ornamental shrub,
shift our focus from natural to synthetic4-5. But it is Bougainvillea glabra Juss.10-11.
noteworthy that using the synthetic dyes has posed Extraction of dyes from their raw dyestuffs is
some serious drawbacks within a period of mere basically a solid-liquid extraction operation. When a
150 years. The research has suspected many of the solid material is brought in contact with a solvent, the
synthetic dyes to release harmful chemicals which are soluble components from solid material move to the
allergic, carcinogenic and detrimental to human solvent. The extraction of plant material results in the
health. About 74 azo dyes have been banned mass transfer of soluble colourant to the solvent. The
worldwide in view of their carcinogenic nature6. rate of mass transfer decreases as the concentration of
dye in the solvent increases, until equilibrium is
___________
*
Correspondent author reached. Thereafter, there is no longer mass transfer
E-mail: [email protected] of dye from plant material to the solvent12.
 PATIL et al: OPTIMIZED EXTRACTION OF DYE FROM BOUGAINVILLEA GLABRA BRACTS 333

One of the steps to improve the mass transfer during optimization. Since its introduction in the U.S.A. in
extraction of dyes from their dyestuff is to investigate early 1980’s, the approach has been popular as
the optimum conditions suitable for efficient extraction. ‘product and process improvement tool’ in the hands
Optimized extraction conditions as suggested by the of engineering and scientific professionals14. The
Taguchi method have been employed to get maximum objective in the extraction of natural dye was to
dye from the source. Further, the mass transfer rates increase its yield through efficient mass transfer
during the extraction have been calculated13. operation. Taguchi method was mainly used to
achieve efficient extraction of dye and to significantly
Materials and Methods reduce the number of experimental trials. The factors
Bougainvillea is a genus of flowering plants native to under investigation included: raw dyestuff particle
South America from Brazil west to Peru and south to size (150, 300 and 425 microns), solid-solvent ratio
southern Argentina. It also grows in tropical and sub (1:20, 1:30 and 1:40), extraction time (60,120 and
tropical forests in India. It is also found in both rural and 180 minutes) and method of assistance for extraction
urban parts of India. It is a common woody shrub that (microwave, enzyme and microwave plus enzyme).
grows to the height of 3-5 m and is sometimes referred There were three levels of each of the factors. These
to as paper flower because the bracts are thin and levels were coded as 0, 1 and 2, respectively.
papery. These bracts can be classic red, purple, orange, Absorbance of the color extract was considered as the
white, pink and cream which bloom all over the year. response variable.
Bougainvillea glabra Juss. (Family- Nyctaginaceae) L9 (34) orthogonal arrays with 4 factors and 3 levels
pink coloured bracts were collected from the road side were used to conduct nine experimental runs15-16. Each
shrubs, separated from their branches and dried in a tray of the nine runs was conducted in duplicate. Other
dryer. The dry bracts mass was ground in a domestic parameters ascertained from the literature survey
mixer-grinder to get mixed particle sizes. The powder included extraction temperature 60°C, speed of the
was sieved to get three different sizes of the dyestuff orbital shaker 150 rpm, microwave power 300 Watt
required for experimental work. with irradiation time 40 second and enzymes
proportion 2% of cellulase and 1% pectinase both on
Chemicals and equipments used weight of raw dyestuff basis17-18. Water was used as a
Cellulase and pectinase enzymes (HIMEDIA solvent in the extraction experiments.
make) were used for enzyme assisted extraction. To Using Minitab software, analysis was conducted
maintain alkaline conditions in extraction medium, for the response variable. The General Linear Model
the buffer tablets (Merck) with pH 9.5 were used. To (GLM) approach was used to perform analysis of
weigh the raw dyestuff and the enzymes, Shimadzu variance and regression of the response variable.
AUX220 Weighing balance was used. Microwave Calculations were performed using a regression
treatment to the raw materials was one of the approach. Conclusions were drawn using calculated
extraction assistance methods. For this purpose, results and the graphs19.
Samsung DE68-02233G domestic microwave oven
was used. Orbital shaker-incubator (Nanolab India Dye extraction with optimized parameters and control
Model NLSIC-23#25/50) was used to carry out the parameters
extraction runs at a time. After the extraction, to The solid-liquid extraction was carried out in an orbital
ensure complete separation of extracted color from shaker incubator using optimum parameters as suggested
the solid waste, syringe and holders assembled with by the Taguchi Design. The particular experiment was
0.2 micron porosity nylon membrane filters named as optimized extraction experiment. Also the
(HIMEDIA make) were used. The absorbance values control experiment was conducted simultaneously with
of the colored extracts were determined by using predetermined control parameters. The control parameters
UV-VIS Spectrophotometer (Shimadzu, Germany, included coarse particle size (425 micron), 1:20 solid to
Model UV1800). FTIR analysis was performed using solvent proportion, three hours of extraction and there was
Cary 630, Agilent Technology spectrophotometer. no any assistance for extraction.
It was aimed to compare control extraction
Taguchi extraction optimization with method of data analysis performance with that of the optimized extraction.
Dr. Genichi Taguchi’s standardized experimental UV-VIS spectra obtained from the two experiments
research design was adopted for dye extraction have been shown in Fig. 1.
334 INDIAN J NAT PROD RESOUR, DECEMBER 2014

Analysis of data which parameter has significant effect on extraction

Statistical analysis and is simply a ratio of the squared deviation to the
The optimization analysis has been shown in mean of squared error. Usually, larger F-value shows
Table 1. The F-value for each parameter indicated greater effect on the extraction value due to the change
of the process parameter. Optimal combination of
process parameters was predicted using ANOVA.

Characterization of the extracted dye

To detect the UV-VIS spectrum profile of the
extract from the source, the extract was scanned in the
wavelength ranging from 200-800 nm by using
UV-VIS spectrophotometer and the characteristic
peaks were detected.
The FTIR spectrum was used to identify the
functional groups of the colourant present in dye
sample based on the peaks values in the region of IR
radiation. The sample was passed into the system and
FTIR Spectrum was recorded in the wavelength
region of 4000 to 650/cm.
The peaks values of FTIR so recorded were used
for functional group analysis. The results of FTIR
analysis supported the presence of colorant in the dye
Fig. 1- UV-VIS Spectra (‫ ג‬max 272nm) sample (Fig. 2).
Table 1- Analysis of variance for absorbance, using adjusted SS for tests
Source DF Seq SS Adj SS Adj MS F P
Replica 1 0.0000 0.0000 0.0000 0.0000 1.000
Particle Size 2 0.0831 0.0831 0.04155 1.70E+05 0.000
Solid-Liquid ratio 2 0.03498 0.03498 0.01749 7.00E+04 0.000
Time of extraction 2 0.0129 0.0129 0.00645 2.60E+04 0.000
Method of assistance 2 0.00914 0.00914 0.00457 1.80E+04 0.000
Error 8 2E-06 2E-06 0
Total 17 0.14011

Fig. 2- FTIR Spectrum of dye from Bougainvillea glabra bracts

 PATIL et al: OPTIMIZED EXTRACTION OF DYE FROM BOUGAINVILLEA GLABRA BRACTS 335

Analysis of data for yield and mass transfer rate calculation The colorant with the molecular weight 551 was
Determination of dye yield:To find out the present in the extract. Therefore, presence of the
concentration of the extracted dye, the dye powder colorant in the extract was 268.42 /551 =0.4871 g/lit
obtained from the spray drier was used to prepare which was 0.04871 g/100 mL. However, 3 g of the
solutions of different concentrations. These solutions raw dyestuff was used for extraction.
were tested for their respective absorbance values
using UV- VIS spectrophotometer. % Yield in the control experiment
The dye concentrations 0.1, 0.2, 0.3, 0.4 and 0.5 = [color extracted/quantity of raw dyestuff]*100
were plotted against the respective absorbance values = [0.04871/3]*100 =1.6238 … (5)
0.02, 0.03, 0.05, 0.08 and 0.09. Fig. 3 showed the For experiment with optimized condition
BEST-FIT line. The slope of this line is the product of c = Absorbance/slope =0.280/0.19 = 1.473 moles/L
path length and molar extinction coefficient. Using … (6)
slope and absorbance, the concentration and the yields
for the control extraction and optimum extraction 0.5 mL of the original extract was diluted to
were determined and compared. 100 mL which is 200 times the original. Therefore the
Beer-Lambert law was used to determine original concentration was 1.473 * 200 =294.73
concentration of an absorbing species in solution. moles/L.
The colorant with the molecular weight 551 was
A = log10 (Io/I) = [ε x L] x [ c ] = slope x [ c ] … (1) present in the extract. Therefore, presence of the
colorant in the extract was 294.73/551 =0.5349 g/L
Thus, Concentration = Absorbance/Slope … (2) which was 0.05349 g/100 mL. However, 3 g of the
raw dyestuff was used for extraction.
The equation of the trend line is
% Yield in the optimized experiment
Y=0.19 X - 0.003 … (3) = [color extracted/quantity of raw dyestuff]*100
= [0.05349/3]*100 =1.783 … (7)
The concentration and the respective yield
calculations are as follows: Mass transfer rate enhancement: Further, using
the absorbance values of the two extraction
For control experiment experiments, the rates of mass transfer were calculated
c = Absorbance/slope =0.255/0.19=1.342 moles/L … (4) and compared. The mass transfer rates expressed in
terms of Absorbance Units (AU) per hour were
0.5 mL of the original extract was diluted to 100ml calculated as below.
which is 200 times the original. Therefore the original The formula for the rate of mass transfer is as
concentration was 1.342 * 200 =268.42 moles/L. follows:
Mass transfer Rate = absorbance/time = A/t, AU/h
For control experiment:
Rate of extraction mass transfer=
absorbance/time=0.255/3=0.085 AU/h … (8)
For experiment with optimum set of parameter:
Rate of extraction mass transfer= absorbance/time
=0.280/1=0.280 AU/h … (9)
When optimized extraction was compared against
control, the enhancement in mass transfer rate was as
follows:
Mass Transfer rate enhancement = Rate (Optimum)/
Rate (Control) … (10)
= 0.280 /0.085
Fig. 3- Absorbance v/s concentration =3.3 times
336 INDIAN J NAT PROD RESOUR, DECEMBER 2014

Fig. 4- Main Effects Plots

Results and Discussion

Optimization results
The optimal combination of the process parameters
was predicted using ANOVA. The statistical analysis
showed the mean values of absorbance as 0.3437,
0.1815 and 0.2302 for levels of particles 0, 1 and 2,
Fig. 5- Structure of betanin
respectively. For solid-liquid ratios with the
levels 0, 1 and 2, the mean absorbance values were chemical functional groups like OH or NH Stretch,
0.3092, 0.2442 and 0.202, respectively. The three carbonyl (O=C), R-NO2 and C-O in the extracted dye.
levels of time of extraction showed the mean The results of FT-IR, UV-VIS spectroscopy as well
absorbance values as 0.2728, 0.2685 and 0.214, support the presence of colourant of the flavones
respectively. Finally for the three respective levels of nature. The molecular formula and molecular weight
the method of assistance, the mean absorbance values (MW) of the colourant are C24H27N2O13 and 551,
shown were 0.2482, 0.281 and 0.2262. respectively. Fig. 5 shows the structure of betanin.
Fig. 4 denoted the optimum set of conditions. The
finest particle size was found as the optimum size. Absorbance, mass transfer rates and dye yields comparison
The optimum solid-solvent ratio found was 1:20. For The absorbance, mass transfer rates and dye yields
one hour of extraction, the absorbance value was the showed significant improvement in case of extraction
highest depicting that the optimum extraction time is with optimized conditions when compared with the
one hour. Extraction enhancement of the colorant was control experiment. It is observed that there was
seen through the highest absorbance in the enzyme significant enhancement in the mass transfer rates.
assistance method. The combined effect of these Amongst the particle sizes, the finest size was the
optimal parameters when used in the extraction of the optimum that gave rise to the increased mass transfer
dye, the absorbance obtained was 0.280 as against the and the increased dye yield. It might be due to the
absorbance 0.255 in the control experiment. highest contact surface area of the finest size. The
optimum solid-solvent ratio was 1:20. Dynamic dye
Colourant, chemical nature transfer equilibrium might have reached with the
The FTIR spectral analysis is important to identify particular ratio. Volume greater than 20 mL was
the characteristic functional groups of the extracted therefore, showing significantly low absorbance
dye. The analysis showed distinct peaks at 3359, values. The complete transfer of dye would have been
2757, 1571, 1011 and 800/cm, respectively. The difficult to the solvent lesser than 20 mL. As regards
broad and strong band at 3359/cm can be attributed to the extraction time, one hour was found as sufficient
bonded -OH groups. The peak at 2757/cm is time for complete extraction. Extraction enhancement
indicative of C-H symmetry in stretching mode. The of the desired component was seen through the
wave number 1571/cm represents the carbonyl group in highest absorbance value in the enzyme method.
stretching mode associated with amide group, the peak Enzyme treatment loosens the structural integrity of
at 1011/cm indicates ether group and finally 800/cm the plant material making their cells easily breakable.
representing p-disubstituted aromatic group. Hence This might be the reason for releasing more solutes
FTIR spectral analysis shows the presence of different from them thereby increasing the mass transfer rate.
 PATIL et al: OPTIMIZED EXTRACTION OF DYE FROM BOUGAINVILLEA GLABRA BRACTS 337

The enhanced mass transfer rate ultimately increases 4 Patil P D, Rao C R and Wasif A I, Natural dyes making their
the yield of the colorant. way back to a golden age, Textile Asia, 2012, 29-31.
5 Haung M, Analysis of EU banned azo colorants in textiles
Thus, the rise in absorbance values, the mass using Poroshell 120 and 1290 Infinity, Agilent Technologies,
transfer rates and the yields might be due to combined Inc., 2011, 1-4.
effect of all the parameters used at their optimal values. 6 Nimkar U and Bhajekar R, Ecological requirements of textile
This has led to the finding that the optimization of industry, Colourage, 2006, 43(4), 135-142.
7 Gokhale S B, Tatiya A U, Bakliwal S R and Fursule R A,
extraction process is beneficial for mass transfer rate Natural dye yielding plants in India, Nat Prod Rad, 2004, 3
enhancement in the natural dye production. (4), 228-234.
8 Mukherjee A, Pollution control in natural colour dyeing
Conclusion through dye strength approximation, Man-Made Textiles in
Optimum extraction conditions investigated by India, 2006, 49(6), 222-225.
9 Venkatasubramanian S, Vijaeeswarri J and Lakshmi Anna J,
Taguchi design were successfully used to conduct Effective natural dye extraction from different plant
natural dye extraction operation in an efficient way. materials using ultrasound, Ind Crop Prod, 2011, 116-122.
The UV-VIS and FTIR results very well supported 10 Sahu N and Saxena J, Phytochemical Analysis of
the presence of betanin colorant in the bracts of Bougainvillea glabra Choisy by FTIR and UV-VIS
Spectroscopic Analysis, Int J Pharm Sci Rev Res, 2013,
Bougainvillea glabra Juss. The results showed 21(1), 196-198.
significant increase in the absorbance of the color 11 Shiv Narayan A K, Sunil Kumar R, Govindasamy S and
extract indicating the significant increase in the mass Chandrasekaran M, Extraction optimization and
transfer rates and therefore, yield of the colorant was characterization of water soluble red purple pigment from
seen to be increased when optimal extraction floral bracts of Bougainvillea glabra, ArabJC
http://dx.doi.org/10.1016/j.arabjc.2013.07.047.
conditions were used for the extraction. This has 12 Treybal R E, Mass Transfer Operations, 3rd edition,
ultimate benefit in terms of reduction in time and McGraw-Hill, New York, 1968, 110-135.
conservation of the resource used for dye production. 13 Sim C C, Kumaresan S and Sarmidi M R, Mass Transfer
The study can certainly contribute to the future coefficients of Eurycoma longifolia batch extraction process,
Proceedings of 18th Symposium of Malaysian Chemical
scale-up of this plant for dye production useful for Engineers’, 2004, 1-20.
textile, food, pharmaceutical, dye-sensitized solar cell 14 The Wealth of India - A Dictionary of Indian Raw Materials
and cosmetic applications. and Industrial Products, Publication and Information
Directorate, CSIR, New Delhi), 1948-1975.
Acknowledgements 15 Montgomery D C, Design and analysis of experiments, 3rd
This study was encouraged by managements of edition, John Wiley and Sons, 2005, 449-562.
16 Patil P D, Rao C R,Wasif A I and Nagla J R, Taguchi
Shivaji University Kolhapur, DKTE’s Textile and optimization for efficient extraction of a natural dye from
Institute, Ichalkaranji and TKIET Warananagar, Bougainvillea glabra bracts, Int J Res Enging & Technol,
Maharashtra State, India. Thanks to all of them. 2014, 03(04), 23-30.
17 Phadke M S, Quality engineering using robust design, 2nd
References edition, Pearson education, Inc., 2009, 35-45.
1 Roy M, Dyes in Ancient and Medieval India, Indian J Hist 18 Wilkins M R, Wilbur W W, Grohmann K and Cameron R G,
Sci, 1978, 13, 83-113. Hydrolysis of grapefruit peel waste with cellulase and
2 Siva R, Status of natural dyes and dye-yielding plants in pectinase enzymes, Bioresour Technol, 2007, 98, 1596-1601.
India, Curr Sci, 2007, 92(7), 916-925. 19 Puri M, Sharma D and Barrow C J, Enzyme-assisted
3 Samanta A and Agrwal P, Application of Natural Dyes on extraction of bio-actives from plants, Trends in Biotechnol,
Textiles, Indian J Fiber & Text Res, 2009, 34, 388-399. 2012, 30(1), 37-44.