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Natural Product Research

Formerly Natural Product Letters

ISSN: (Print) (Online) Journal homepage: www.tandfonline.com/journals/gnpl20

Natural colour extraction from horticultural crops,

advancements, and applications—a review

Gokiladevi R, P. Ellampirai M, Ramesh Kumar A, Srivignesh S & K. Rama

Krishna

To cite this article: Gokiladevi R, P. Ellampirai M, Ramesh Kumar A, Srivignesh S & K. Rama
Krishna (17 Nov 2023): Natural colour extraction from horticultural crops, advancements, and
applications—a review, Natural Product Research, DOI: 10.1080/14786419.2023.2280796

To link to this article: https://doi.org/10.1080/14786419.2023.2280796

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Published online: 17 Nov 2023.

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Natural Product Research

https://doi.org/10.1080/14786419.2023.2280796

Natural colour extraction from horticultural crops,

advancements, and applications—a review
Gokiladevi R, P. Ellampirai M, Ramesh Kumar A, Srivignesh S and K. Rama
Krishna
Department of Horticulture, School of Life Sciences, Central University of Tamil Nadu, Thiruvarur, India

ABSTRACT ARTICLE HISTORY

The scope for natural colours is increasing because of the aware- Received 20 July 2023
ness of allergic, toxic, and hazardous reactions associated with syn- Accepted 2 November
thetic dyes. Natural colours are extracted from sources that are 2023
naturally available, such as flora, fauna, and minerals. Nature KEYWORDS
sourced us multiple possibilities of colours with varied shades and Supercritical; micro-wave;
hues that are subtle and harmonious when combined. Reasons like pharmaceutical; food
the instability of natural colours during industrial processing, sea- colourants; textile;
sonal availability of raw materials, fading of colour over time, cost intelligent-packaging
of the benefit, and struggles in attaining sustainability have
reduced commercialisation success as synthetic colours. Some
plants that yield natural colours are also included in crop rotation
practice. Natural dye extraction is a source of employment for the
countrified subdivisions of poor developing countries. Indigenous
technologies on natural colour extraction are available and have
been practiced over the years; due to a lack of documentation and
information on colour-yielding plants or products from horticul-
tural crops, and their extraction methods, the use of natural colours
is diminishing day by day. Even in recent years, emerging tech-
niques have been adopted in research and development, and the
information has not been brought together for the use of indus-
tries and allied sectors. Several modern approaches, such as
Ultrasonication, microwave, enzymatic, supercritical, pressurised
liquid extraction, etc., have proven to give better results in extract-
ing natural colours. Thereby, having instantaneous information will
help to go green, be eco-friendly, and effectively utilise all the
resources without compromising industrial benefits. Reviewing the
availability of natural colours from horticultural crops, classifica-
tions, recent trends in their extraction process, and applications in
various fields will help achieve the above.

CONTACT K. Rama Krishna ramakrishna.karri001@gmail.com

Supplemental data for this article is available online at https://doi.org/10.1080/14786419.2023.2280796
© 2023 Informa UK Limited, trading as Taylor & Francis Group
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2 G. R ET AL.

1. Introduction
Colours have played an essential part in the industrial revolution. Before that, colours
were obtained from natural resources, such as plants, animals, and minerals. After
the industrial revolution, there was high demand for colours in sectors, such as textile,
food, pharmaceuticals, cosmetics, and fashion, etc. which led to immense production
of synthetic dyes with the added advantage of low price (Kumaresan et al. 2011).
Thereby the slow diminishing of natural colours due to their scarce availability and
high cost. The global natural food colours market was valued at U.S. dollars (USD)
1.54 billion in 2021 and is expected to grow at a compound annual growth rate
(CAGR) of 7.4% during the forecast period (Polaris Market Search 2023). As per Market.
us, the food colourants market size is projected to surpass around USD 10 Billion by
2032, and it is poised to reach a CAGR of 9.3% from 2023 to 2032. According to
Dikshit and Tallapragada (2018), Food colourant estimated market share is ~41% for
synthetic colour, 20% for Natural identical colours, 11% for Caramel colours, and 27%
for Natural colours.
The use of natural colour is growing at a faster rate due to strict regulation standards
followed by many countries to reduce the hazardous and carcinogenic effects of syn-
thetic colours; the use of natural colours has recently gained attention. Due to their
non-carcinogenic, non-toxic, and biodegradable nature, the natural colours have been
considered safe for the environment and humans (Cristea et al. 2003). The rich biodi-
versity of India (sub-continent) provides plenty of raw materials originating from nature
sources, especially horticultural crops, such as fruits, vegetables, flowers, spices, etc.
India has recently achieved new heights in the production of horticultural crops (310.73
mMT), giving enormous opportunities for utilisation in natural colour extraction as
horticultural crops are a rich source of natural colours. To achieve a continuous supply
of natural colours to industries, we need to develop a sustainable link chain between
cultivation, collection, and their use (Gokhale et al. 2004). Even though India has
achieved higher production in natural colour-yielding crops, it is enabled to cater to
the needs of industries and societies where colours play a pivotal role in their growth.
The setback in utilising natural colours from horticultural crops may be due to the lack
of awareness of their source, extraction, and application. In most developing countries,
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Natural Product Research 3

natural colours are used on a small scale compared to commercial synthetic colours.
However, with the increasing worldwide concern over the environment, eco-friendly
natural colours have gained interest once again (Agarwal and Tiwari 1989). The review
provides holistic scientific information on the availability, sources, applications, and
techniques of extracting natural colours from the horticulture sector. This information
can be utilised for improving production, thereby sustaining the growth of natural
colours linked business and reducing the dependency on synthetic dyes.

2. History and classification of natural colours

The history of natural colourants was identified in 2600 BC. When it comes to India,
the Indus Valley civilisation was the pioneer of using natural pigments during 2500
BC. This was evident from the finding of the madder-dyed coloured garments at the
excavation site of Harappa and Mohenjodaro during 3500 BC. During 300 BC, artificial
colours were used in the beverages like wine. Between the period of 1850–1900,
synthetic dyes became very common all around the world. In earlier periods, natural
colour classification was done based on the alphabetical order of their scientific names
(Gulrajani et al. 1992). Later the classification was done based on their origin, hues,
chemical constitution, and applications (Paul, 1996) (Figure S1). The details of the
classification of natural colours are presented in the Supplementary File 1.

3. Trending applications of natural colours

3.1. The textile industry and UV protective finishing
The evidence of textile dyeing was found 5000 years ago at Mohenjodaro, Pakistan.
Using suitable finishing agents and mordant is the most desirable customer choice.
The best choice for dyeing wool, leather, and cotton is madder extract. Due to the
different chemical constituents of natural colours, the dyeing procedure will differ
from the synthetic dyeing process. When anthraquinone dyes complex with metal
salts, it will increase the fastness of the colour (Shahid et al. 2013). Using madder,
weld plants, and indigo for the production of natural colours has been identified
as an economically feasible technique in terms of raw materials, production, and
waste handling cost (Sirait 2018). The application of natural dye in textiles can
shorten production time and cost. The optimisation of natural colour production
can be done by combining the utilised waste from natural colour extraction with
the extract obtained from cultivated plants (Shahid et al. 2013). Ultraviolet radiation
from the sun causes many skin problems like sun burn, skin tanning, premature
skin ageing, wrinkles, and overexposure can even lead to skin cancer. Using pigment,
delustrants, or ultra violet absorber quality containing natural colours in textile will
block the sun rays and protect the skin from damage (Table S2). It implies that
dyed fabrics protect more than un-dyed fabrics. It was also noticed that protection
level is directly correlated with the concentration of natural colour used. Due to the
environmental and energy conservation benefits that are associated with natural
colours, textile industries have a promising future for UV-protective fabrics. UV pro-
tection capacity of the dyed fabrics depends on the porosity, thickness, moisture
content, structure, and chemical nature of fabrics (Gies et al. 2007). The UV protection
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4 G. R ET AL.

property of dyed fabrics has increased with metallic salts (Gulrajani and Deepti
2011). Recently, it was noticed that extraction from the orange peel would have the
fantastic UV-protecting property (Hou et al. 2013). According to Zhou and Tang
(2016), silk fabric coloured with water-soluble curcumin extracted from the root of
Curcuma longa L was compared with the silk fabric coloured with modified curcumin
(a water-soluble reactive UV absorber was introduced into curcumin molecule), and
both were tested. The silk fabric dyed with modified curcumin had excellent colour
fastness, extremely good UV protection ability, high antibacterial activity comparable
to the curcumin dyed silk, but its antioxidant activity was inferior than cucurmin
coloued fabric.

3.2. Food colourant

Due to the benefits of quality and health promotion that are associated with nat-
ural colours, both the manufacturers and consumers prefer natural colours in place
of synthetic ones (Dawson 2008). Natural colours produce hues ranging from green
through orange, red, blue, yellow, and violet, depending on the stability of the
colourants used. Natural colourants extracted from horticultural crops had shown
an anti-parasitic effect, which was reflected when ethanolic and aqueous extract
from Vitis vinifera L. were prepared and tested for the anti-leishmanial activity
against the promastigotes (Leishmani ainfantum). The result showed that the eth-
anolic extract is more active than the aqueous one (IC50 = 0.108 mg/mL) at the
inhibitory concentration 50(IC50). The ethanolic extract under microscopic obser-
vations showed that it boosted the destruction of membranes (Cytoplasm and
nuclear) of promastigotes and its shape of the cell. The enhanced anti-leishmanial
activity of ethanolic extract is due to the presence of more decadent amount of
anthocyanin when compared to aqueous extract (Mansour et al. 2013). Saffron
extract (50 mg) tested for its antioxidants property in wheat cookies showed that
cookies with saffron extract had reduced spread ratio, hardness, and increased
Colour values, with higher retention of antioxidant properties, sensory scores, and
overall acceptance upto six months of storage (Bhat et al. 2018). Recently fungi
are used as cell factories for natural colour production by using biotechnology.
Intervention of biotechnology has led to mass production of fungal cells (Delgado
Vargas et al. 2000) and the colours produced from them are effective antioxidants
and pro-anthocyanins. They are also effective in increasing the shelf life of food
and had positive affect on consumer health. Lin et al. 2018, had used Gardenia
Yellow (G), Curcumin(C), Soy sauce (S), and Radish Red (R) as a natural food colou-
rant in dried-tofu. The findings demonstrated that the formulas G:R = 0.2:0.8 and
C:R = 0.08:0.92 as mixed natural colourants had the same eye sensory quality
approval and the intention to purchase by the consumers as that of synthetic
colour. Furthermore, it was indicated that dietary exposure to the level of meth-
ylglyoxal (50 M) found in soy sauce did not significantly alter the cellular shape
and cell cycle of neurons. Overall, the soy sauce as a colouring agent was safe,
thereby demonstrating its potential as a food colourant. Some of the natural colours
and their nutritional values are given in Table S3.
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Natural Product Research 5

3.3. Pharmaceutical colours

For using natural colours in medicinal products, an additional licence is required. Its
addition may impart other benefits to the medicine. Using arjuna or sariva will thin
the blood, reduce the RBC agglutination, and purify the blood. Flavonoids have shown
reduced vascular purpurea incidence and strengthen the capillary walls. Anthocyanin
will help aid vision and reduce cataract incidence (Aggarwal 2021) (Table S4). The
chemopreventive potential from the leaf powder of henna (Lawsonia inermis) and law-
sone extracted from it was evaluated for its inhibition effect of Epstein-Barr virus early
antigen (EBV-EA) in mice through oral and topical. Feeding of henna powder (0.0025%)
through drinking water reduced tumour incidence by 66% and lawsone (0.0025%) by
72% (Kapadia et al. 2013). In Another study, where Black tea’s theaflavin monomers,
such as theaflavin 3gallate (TF2a) and theaflavin 3′-gallate (TF2b) had significantly
inhibited the growth of cisplatin-resistant ovarian cancer A2780/CP70 cells while being
less cytotoxic to the normal ovarian IOSE-364 cell line. The study found that TF2a and
TF2b could treat and prevent platinum-resistant ovarian cancer (Pan et al. 2017).

3.4. Beauty aids

Increasing awareness among consumers of the side effects associated with synthetic
dyes, the market for natural colourants in cosmetics has increased. They are used in
foundations, hair colouring, lip care products, colour cosmetics, including UV protec-
tion and anti-ageing. Sources of colours that are used in cosmetics are turmeric,
saffron, paprika, coffee, tea, beetroot, pumpkin, calendula, etc., In earlier period,
synthetic dyes were used as colourants in cosmetics (Table S5). The harmful carcino-
genic and toxic effects of synthetic dyes have created thirst for using natural colours.
Due to the anticancer and antioxidant properties of natural colours, synthetic colour
in the cosmetic industry has now been replaced by natural colours (Mohana Priya
et al. 2020). Annatto (extract from Bixa orellana L.), β-Carotene from fruits, and henna
are extensively used in the cosmetic industry (Weisz et al. 2007). Rosa rubiginosa, a
flower of the Bougainvillaea spectabilis plant, Beetroot (Beta vulgaris), and Crocus sativus
flower were assessed for their utility as cosmetic lipstick based on factors including
melting point, breaking point, a force of application, surface anomalies, ageing sta-
bility, perfume, etc. It was determined that herbal lipstick had a low risk of side effects
with the greatest local impact on the lips (Mahanthesh et al. 2020).

3.5. Therapeutic uses

Natural colours, in addition to dyeing properties, also have a wide range of medicinal
uses. Antioxidant and anti-inflammatory benefits associated with anthocyanin and
carotenoids are used in therapeutic theory. Using natural colour as therapeutic therapy
is as ancient as human civilisation. Colours from pomegranate, lawsone from henna,
and juglone from walnut possess antibacterial and antifungal properties. Combining
Colour therapy with conventional medication and physical therapy has been proven
to have a positive impact on improving the condition of a patient with generalised
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6 G. R ET AL.

anxiety disorder (GAD) and depression. It also reduces negative emotions, improving
positive emotions than conventional treatment (Samuel et al. 2022). Indicaxanthin is
a naturally occurring colouring compound in Opuntia ficus-indica (prickly pear) (Allegra
et al. 2019). Indicaxanthin pigment is a novel therapeutic drug that has to be studied
in more intricate combination therapy studies by pairing it with other therapeutic
agents that target various disease progressions.

3.6. Insect repellent

Insects cause significant damage to textile products, such as carpets, garments, blankets,
and handlooms during warehouse storage. Eight of the ten insects tested for their
insect-repellent properties performed well against Anthrenus verbasci. It was in the
order of ‘lac dye, gall nut, catechu, red cabbage, cochineal, indigo, Amur cork tree
extract’. Cochineal and madder-containing anthraquinone compounds were found very
useful in protecting wool fabrics against black carpet beetles. This study was done by
Kato (Kato et al. 2004). The lowest weight loss and the best effect of insect deterrence
was witnessed in the use of madder dye. Example: identification of anti-insect properties
from lichens (Rangno and Hobbs 2005). Selective natural colourants showed functional
properties where it was discovered that wool fabric dyed with henna (L. inermis) and
walnut shell (Juglans regia) extracts showed a mothproof effect (Nazari 2017).

3.7. Intelligent packaging

Active and intelligent packaging are new concepts that have largely contributed
to safer and healthier food. According to the European Commission [Commission
Regulation (EU) 1935/2004; Commission Regulation (EU) 450/2009], active packaging
materials are ‘materials and articles that are intended to extend the shelf-life or
to maintain or improve the condition of packaged food which are designed to
deliberately incorporate components that would release or absorb substances into
or from the packaged food or the environment surrounding the food’. Due to their
easy preparation, renewability, low toxicity, and eco-friendly properties, natural
colourants play a significant role in smart packaging. The utilisation of natural
colourant in food packaging technologies to improve food properties, extend shelf
life, and monitor food freshness was demonstrated by using colorimetric indicators
that sense the changes in the pH of the foodstuff and exhibit colour change can
enable real-time monitoring system of food quality (Echegaray et al. 2023). Natural
dyes β-carotene, curcumin, and lutein were applied as indicator substances in
intelligent biodegradable packaging materials and showed a definite change in
colour under the influence of elevated temperature, weathering, and UV radiation
(Latos-Brozio and Masek 2020). Pourjavaher et al. (2017) created intelligent pack-
aging using bacterial cellulose nanofibers that were loaded with concentrated and
diluted anthocyanins from purple cabbage. The Films containing more anthocyanin
in various pH solutions did not altered significantly, particularly at pH values higher
than 5. The film with diluted anthocyanins displayed a more apparent reaction to
pH between 2 and 10, which may be an indicator of pH changes in packed food.
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Natural Product Research 7

3.8. Dye-sensitised solar cell

Solar energy is one of the renewable energies that is promisingly eco-friendly. The
solar cells are capable of converting solar energy into electrical energy. Dye-sensitised
solar cells (DSSC) are innovative solar cells that perform a similar process to photo-
synthesis in plants. The natural colour as a sensitiser plays a crucial role in absorbing
sunlight and transforming solar energy into electrical energy (Zhou et al. 2011). The
ruthenium-based dyes are extensively used in DSSC (Campbell et al. 2007). Researchers
are searching for alternatives to ruthenium-based dyes because of their high cost,
limited availability, and unacceptable environmental Impacts (Campbell et al. 2007).
From many types of research, it was found that natural colours like betalains, chlo-
rophyll, chalcone, and anthocyanins are also effective when used as an alternative to
ruthenium (Campbell et al. 2007; Chang et al. 2010; Narayan 2012). Natural dyes were
taken from flowers of Amaranthus caudatus, Bougainvillaea spectabilis, Delonix regia,
Nerium oleander, Spathodea companulata, and a combination of the extracts to create
dye-sensitised solar cells (DSSCs) which were investigated for the PEC performances
of DSSCs using mixed dye solutions.
In contrast to the individual extracts, the combined extract does not exhibit any
synergistic photosensitisation. The best performance was instead displayed by the
cell sensitised with ethanol extract of the A. caudatus flower alone. The instruments
revealed that the incident monochromatic photon to current conversion efficiency
(IPCE) ranged from 4.7 to 52%. The results from the IPCE data and the (J-V) curves
agree with each other (Godibo et al. 2015).

4. Extraction methods
Phytochemicals are naturally occurring substances that are found in plants. The.
Secondary metabolites, such as Alkaloids, Flavonoids, Tannins, Phenolics, saponins,
Steroids, glycosides, and Terpenes help protect human health and plants from diseases.
This phytochemical are primarily found in fruits, vegetables, flowers, grains, nuts,
seeds, herbs, and spices. The extraction method is essential for analysing the phyto-
chemicals. Among several phytochemicals, natural colours found in the fauna and
flora contribute majorly to the well-being of humans and impact the socio-economic
status. The traditional methods for these colours include maceration, Soxhlet extraction,
Aqueous extraction, and Solvent extraction (Acid, alkali, and alcoholic) (Figure 1). The
advanced extraction methods, such as Supercritical fluid extraction, Microwave-assisted
extraction, Ultrasound-assisted extraction, fermentation, and Pressurised liquid
extraction, are presented below (Table 1).

4.1. Maceration
In this method, the plant parts are cleaned, dried, and made into powder form. The
powdered plant material is kept under the contact of solvent in a stoppered container
for a defined period with continuous agitation until soluble material is dissolved. It
is the first step for the extraction of colours and is influenced by pH and temperature.
An anthocyanin dye was extracted from red dragon fruit peel by macerating the
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8 G. R ET AL.

Figure 1. Modern extraction methods.

sample for four days using 10% citric acid solution (1:3), yielding 22.92% dye con-
taining anthocyanin levels of 52.27 mg/L with superior L, a, b, hue values and the
best anthocyanin dye stability was obtained at a pH of 3 and a temperature of 7 °C
(Dewi et al. 2020).

4.2. Soxhlet extraction

The dried plant parts were put into the thistle of the Soxhlet extractor, and methanol
was used as a solvent. Temperature is maintained. Several cycles of solvent were run
to extract all the compounds from the plant parts. The extract obtained from the
carrot through the Soxhlet extraction method contained a higher amount of beta
carotene and total carotenoids when compared to the microwave-assisted extraction
method. A better amount of bioactive contents (beta carotene and total carotenoids)
can be obtained by blanching carrots in water (29.74 and 58.04 mg/100 g dry basis)
and in citric acid (32.08 and 61.62 mg/100 g dry basis), respectively (Hiranvarachat
et al. 2013).

4.3. Aqueous extraction

This method was traditionally used to extract natural colour from plants and other
substances. The natural colour-containing material is first broken into small pieces or
sieved or powdered to improve the efficiency of extraction. Then, the powdered
material is soaked with water in the wooden, earthen, or metal vessels for a longer
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Table 1. Extraction of colour compounds from horticultural crop sources.

Extracted
Extraction pigments and
Crop name techniques Operation works yield References
Blackberry Soxhlet extraction Temperature-80 °C, Anthocyanin Machado et al.
solvent-methanol, time-300 min (1.33) 2015
Carrot Soxhlet extraction Temperature-58 °C, solvent-mixed Beta-carotene Hiranvarachat
solvent (acetone, hexane, (0.29) et al. 2013
ethanol), time-360 min
Mango Maceration Solvent-ethanol, methanol, water Xanthone Sutrisna et al.
2020
Beetroot Microwave assisted Solvent-ethanol, microwave Betanins (0.19) Cardoso-Ugarte
extraction power-400 W, time-3.50 min et al. 2014
Carrot Microwave assisted Temperature-58 °C, solvent-mixed Beta-carotene Hiranvarachat
extraction solvent (acetone, hexane, (0.23) et al. 2013
ethanol), microwave
power-180 W, time-3 min
Bougainvillaea Ultrasonic assisted Temperature-55 °C, Betacyanin (1.72), Maran et al. 2015
extraction solvent-methanol, power-88 W, betaxanthin
frequency-20 KHz, time-37 min (5.78)
Marigold Enzyme assisted Enzyme: viscozyme, Carotenoids (0.5) Barzana et al.
extraction time-60 min,temperature-25 °C, 2002
solvent-hexane
Green elfcup Liquid 70 °C and 15 min incubation Xylendin Zschätzsch et al.
(mushroom) fermentation 2021
Sulphur polypore Liquid 70 °C and 15 min incubation Laetiporic acid Zschätzsch et al.
(mushroom) fermentation 2021
Carrot Supercritical CO2 Temperature: 60 °C, pressure: 1–2% β-carotene Klettenhammer
extraction 30 Mpa, solvent: CO2, flow rate: et al. 2021
2 L/h, time: 30 min
Black berry PLE Temperature: 100 °C, pressure: Anthocyanins Machado et al.
75 bar, solvent: water with 50% (1.02) 2015
(v/v) ethanol, flow rate:
3.35 mL/min, time: 30 min
Dunaliella salina SFE Temperature: 60 °C, pressure: Carotenoids Macías-Sánchez
300 bar, solvent: CO2, flow rate: (14.92), et al. 2009
4.5 mmol/min, time: 180 min chlorophylls
(0.27)

time, which helps to lose the cell structure, and then boiled to get the plant extract,
followed by filtering to remove the debris. The boiling and filtering process had to
be repeated to get more amount of coloured materials from the extract. Most of the
dyeing operations are carried out by aqueous media for easy application of dye to
the textile materials. The major disadvantages of this extraction method are long
extraction time, more water requirement, and usage of higher temperatures. The
seeds of Cassiva tora and Grewia optiva, as well as the leaves of a eucalyptus hybrid,
were successfully used by Dayal and Dobhal (2001) to extract an aqueous solution.
Similarly, the aqueous extraction was used potentially in natural dyes extracted from
a variety of plant sources, such as coffee seed, tea, onions, tamarind (Tamarindus
indica), henna (Lawsonia inermis), lemongrass (Cymbopogon citratus), cardamom
(Elettaria cardamomum), beetroot.

4.4. Solvent extraction (acid, alkali, and alcoholic method)

Many plant-coloured extracts are in the form of glycosides, so acidic or alkaline
methods can extract them. The addition of acid/alkaline helps to facilitate the
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10 G. R ET AL.

hydrolysis of glycosides, resulting in good extraction and a high yield of colouring

materials. In Butea monosperma flowers, an acid method of extraction is used. The
Alkali method is used for the phenolic group of secondary metabolites as they are
soluble in alkali. The disadvantage of this method is that the colouring materials get
destroyed under alkaline conditions because of the sensitivity of pH. Natural colour
extraction from Acacia catechu using an alkaline medium (NaOH 20 g/L) for imparting
brown colour to silk was standardised where the colour was extracted by boiling the
Khair powder at pH 12 and 90 °C for 120 min (Khatun et al. 2017). The best extraction
method for the Siris bark in powder form is determined using four types of extraction
mediums (aqueous, acid, alkali, alcoholic) with varying solvent ratio, time, and
material-to-liquor ratio. The optimum condition for extraction is a solvent ratio of
30% alcohol at 5 h of extraction time and material to liquor ratio of 1:10 (Thanabal
et al. 2021).

4.5. Ultrasonic-assisted extraction

The efficiency of extraction is increased by using ultrasound or microwave, which
helps to reduce the quantity of solvent, temperature, and time of extraction. The
plant material are treated with any solvent under the presence of ultrasound, which
creates small bubbles in the medium. As the exposure time increases, the size of the
bubbles gets increased, thereby unable to retain their shape and leading to the
busting of the bubbles, creating a high temperature and pressure. This will ease the
extraction process, increasing the extraction efficiency within a short period of time.
Extraction of pigments from bougainvillaea flower using ultrasonic-assisted extraction
(UAE) by adopting four factors three level Box-Behnken response surface design has
shown that the optimised level of temperature (55 °C), 88 W ultrasonic power, 37 min
extraction time, and SL ratio of 1:17 g/mL were ideal for extraction of betacyanin
colour from bougainvillaea flower (Maran et al. 2015). Adeel et al. 2019 isolated tannin
from neem bark with the help of chemical and biomordant using ultrasonic (US)
treatment and it was tested on silk fabric. It was found that US-treated neem bark
for 30 min had yielded methanolic extract of 5PH (8 g dye powder) and gave high
colour strength if used on dye irradiated (65 min at 75 °C) silk. It was concluded that
US-based isolated colourant gave excellent characteristics using bio-mordants onto
silk fabrics.

4.6. Microwave extraction

In microwave extraction, the natural extract was treated with any solvent under
microwave energy. This method requires less extraction time with higher extraction
efficacy. Sinha et al. 2013 reported the extraction of annatto colour with the help of
microwave energy sources. Microwave-assisted extraction of betalins from diced beet
roots exhibited that the higher betanine yield was obtained at the combination of
400 W and 100% duty cycle for 90–120 s, whereas higher betaxanthins were obtained
at 140–150 s. As compared to the conventional extraction method at 80 °C, MAE
attained twice the amount of betalain yield (Cardoso-Ugarte et al. 2014). The extraction
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Natural Product Research 11

of colourant from the pomegranate rind using a microwave-assisted method showed

that the extraction efficiency of the colour extract (13–100%) was obtained at opti-
mum conditions of dye extraction time of the 90s and pH 3.5 (Sinha et al. 2012).
Adeel et al. 2023 reported that the extraction of tannin-based natural dye from waste
tea leaves under the influence of microwave treatment (6 min) along with the use of
bio mordants (4% acacia extract, 1% pomegranate extract, and 5% turmeric extract
as pre-bio-mordant, whereas 5% of acacia and 2% of pomegranate and turmeric as
post-bio-mordants) was tested on microwave treated wool at 80 °C. The results showed
that adding pre and post-bio mordant along with microwave treatment has not only
improved the colour strength of tannin dye onto wool fabric but also made the
process sustainable.

4.7. Fermentation
Fermentation uses the enzymes which are produced by the micro-organisms. Indigo
extraction is an example of this method. Harvested Indigo leaves and twigs are soaked
in warm water and allowed for fermentation. After completion of fermentation, the
plant material is collected, washed, and then pressed into cakes. The fermentation
extraction method is similar to the aqueous extraction method, except that this
method does not require high temperature. The disadvantages of this method are
longer extraction time and foul smell due to the action of microbes. The primary
mechanism behind the indigo fermentation under alkaline conditions was driven by
indigo-reducing bacteria. Due to the absence of oxygen in the fermentation system,
the count of anaerobic bacteria dominates the aerobic one, which leads to the initi-
ation of staining in the textiles. Many species of bacteria are capable of indigo
reduction, but the performance was based on the substitution of bacteria. Under
anaerobic conditions, the fermentation can last for some six months, which favour
contamination to happen. So, neutral natural fermentation is a very much suitable
technique for indigo fermentation (Aino et al. 2018). The biomass fermentation of the
indigo plant for 12h at a pH range of 6–6.8 and temperature of 40 °C yielded maxi-
mum pure indigo (2.84 mg g−1) (Dutta et al. 2017).

4.8. Enzymatic extraction

The enzymes, such as cellulase, pectinase, and amylase are used to loosen the mate-
rials surrounding the cells, which leads to the proper extraction of natural colours.
This extraction method helps to extract the colour from hard plant materials, such
as bark, roots, etc. The enzymatic extraction of colour from milled fresh flowers of
marigold using Viscozyme (cocktail + polysaccharases) recovered 97% of carotenoids
when tested at an 80 L scale (Barzana et al. 2002) under constant conditions. Shen
et al. 2021 have done response surface method (RSM)-based optimisation for cocktail
enzyme-assisted extraction of jujube peel pigments. The ideal extraction conditions
for the highest quantities of (TPC), (TFC), and a* values were 16 mL BLV, pH 7.0, 43 °C,
and 97 min of incubation.
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12 G. R ET AL.

4.9. Supercritical carbon dioxide extraction (ScCO2)

It is a process of separating one component from another component using super-
critical fluids as a solvent. This is an advanced separation technique based on the
power of gases above the critical point. The preferred gas is carbon-dioxide because
of its lower critical temperature and non-toxic, non-flammable, and low cost. Carbon
dioxide can be used at a temperature of 40–80 °C and pressure of 35–70 MPa. The
advantage of this method is the extract is free from residual solvent traces and heavy
metals. The process of extraction become popularised in the extraction of natural
products for food and pharmaceutical applications. The disadvantages of this method
are the higher cost of equipment and poor extraction of polar substances.
The raw material studied was Dunaliella salina for carotenoid and chlorophyll
extraction, CO2 as a solvent, and the supercritical extraction process is more suitable
for the higher recovery of carotenoids at 40 and 50 °C at 400 bar and chlorophyll
(60 °C at 500 bar) when compared to the conventional technique (Macías-Sánchez
et al. 2009). The mango airdried peel was subjected to ScCO2, where higher yields of
carotenoids were found at 25.0 MPa, 60 °C, and 15% w/w ethanol (Sánchez-Camargo
et al. 2019). Using supercritical carbon dioxide (SC-CO2), the natural red pigment
anthocyanins were extracted from roselle calyces. Maximum anthocyanin, phenolic,
and flavonoid content was achieved in the optimal circumstances of 27 MPa, 58 °C,
and 8.86% co-solvent ratio, with low L* and C* values. Stored Anthocyanin content
obtained from SCCO2 was tested for its average reaction rate (k) and half-life (t1/2)
and compared with extract obtained from Solid-Liquid extraction (SLE). Findings
showed that compared to traditional SLE techniques, the examined conditions using
SC-CO2 gave lower degradation rates and a longer t1/2 (Idham et al. 2022).

4.10. Pressurised liquid extraction

Pressurised liquid extraction, otherwise known as accelerated solvent extraction, has
been popularised for the last decades. In this method, acetone/hexane and toluene
(organic solvents) have been used as solvents. This method is exclusively used for
extraction only. Pressurised liquid extraction was one of the rapid and clean techniques
among the conventional techniques of pigment extraction available in the industry
for the extraction of bioactive compounds like anthocyanin from blackberry and other
residues of fruits and vegetables. To get the maximum yield, the polarity of the sol-
vent and temperature should be under control. The blackberry fruit samples were
subjected to pressurised liquid extraction, under which a higher amount of anthocy-
anin can be obtained from the blackberries were the combination of ethanol + water
(50%V/V) at the temperature of 100 °C under 7.5 MPa (Machado et al. 2015). Pressurised
liquid extraction (PLE) was used to extract grape marc to produce extracts high in
monomeric anthocyanins (MAC) and total phenolic compounds (TPC). At temperatures
ranging from 40 to 100 °C, PLE was carried out using combinations of ethanol and
water (acidified or not). The optimal PLE conditions for MAC extraction [10.21 mg of
malvidin-3-O-glucoside/g of dried grape marc (dr)] were ethanol-water pH 2.0 [50%
w/w] (Pereira et al. 2019).
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Natural Product Research 13

5. Conclusion
Natural colours are non-toxic and act as a sustainable resource in the environment.
Natural colours provide an environmentally safe colouring of food and other materials.
Colour is the crucial point of attraction from consumer point of view for their accept-
ability of the product. Industries are now using natural colours as the consumer
demand for natural pigmentation has increased because of their non-toxicity, envi-
ronmentally friendly, and health benefits. The improved extraction methods aided in
expanding the applications of natural colours due to their cost effectiveness, thereby
plummeting the challenges in colour industry and its associates. The improved tech-
nological advances in colour extraction also show their efficiency, a pivot important
as the natural resources are at stake. Therefore, having instantaneous information will
help to go green, be eco-friendly, and utilise all the resources in an effective manner
without compromising industrial benefits.

Disclosure statement
No potential conflict of interest was reported by the author(s).

Funding
The author(s) reported there is no funding associated with the work featured in this article.

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