Discover Sustainability

Review

Floral bio‑residues of saffron: a potential source of valuable

components; extraction and application
M. Anjaly Shanker1 · Abhilash Puthiyedath2 · Sandeep Singh Rana1

Received: 2 August 2024 / Accepted: 10 February 2025

© The Author(s) 2025  OPEN

Abstract
Prevalently known as red gold, saffron is one of the popular and expensive spices throughout the world. Assortment
of the spice is an energy-intensive process where the stigma part of the flower is separated as a primary resource.
Structurally, the stigma accounts for the small part of the flower and the remaining major proportions including petals,
stamen, style etc. are discarded as waste fractions. With saffron being known for the culinary, medicinal and functional
properties, there is higher value attributed to the waste fractions of the flower. Bio-fractions are known sources of pro-
tein (8.17–24.45 g/100 g of dry weight), lipids (2.22–10.73 g/100 g of dry weight), carbohydrates (33.8–64.9 g/100 g of
dry weight), minerals and dietary fractions (14.4–32.3 g) and are highly comparable with the nutritional percentages of
stigma. Phytochemical profiles of these parts are prominent with higher percentages of carotenoids, flavanols and antho-
cyanins making them a valuable extraction source. Maceration and solvent extraction are traditional methods employed
in the extraction of these plant components. The involvement of modern extraction methods like ultrasound, micro-
wave, supercritical extraction techniques have increased the efficiency and functional quality of the extract obtained.
This review aims at assessing the phytochemical and nutritional profile of these bio fractions, emphasizing the positives
of different extraction techniques and also covering the unlimited potential applications of so-called waste fractions.

Article Highlights
• Waste fractions from saffron production are rich in bioactive and functional components and are comparable with
saffron.
• Component and functional profile ensure profound application possibilities of these bio-fractions especially, in the
pharmaceutical, cosmetic and food industry.
• Innovative and green extraction techniques were advantageous over conventional extraction techniques, ensuring
quality and efficacy of the final product.

Keywords Saffron bio-residues · Phytochemical activity · Green extraction techniques

Abbreviations
DF Dietary fibre
RH Relative humidity
GAE Gallic acid equivalent

* Sandeep Singh Rana, [Link]@[Link] | 1Department of Bio Sciences, School of Bio Science and Technology (SBST), Vellore
Institute of Technology, Vellore 632014, India. 2Manne Kancor Ingredients Pvt. Ltd, Angamaly South, Ernakulam 683573, India.

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FRAP Ferric reducing antioxidant power

QE Quercetin equivalent
SCF Supercritical fluid
LC Lethal concentration
IC Inhibition concentration

1 Introduction

Waste management is the major problem encountered throughout the globe, and its composition diversity is something
that limits their ways of reduction. The agri-food processing sector has been identified as a major contributor towards this
environmental burden, generating vast amounts of waste materials into the system. The residual mass from the process-
ing wastes is characterized as biomass rich in moisture and microbial loads, imperiling the environmental conditioning
and stability. Landfilling and incineration are the common methods involved in the process of disposal but are aligned
with hazardous, toxic, and unfavourable aftereffects to the system and the environment [1]. Management and produc-
tion pathways involving zero or minimum waste is one of the major challenges associated with the agri-food sector
and the stakeholders involved in food production. Studies on these biomasses have reported the presence of different
bioactive components like polyphenols, essential oils, carotenoids etc., delivering them as an efficient, cost-effective
and sustainable raw material [2]. This is the reason behind the efforts to involve and use all the byproduct elements in
the major line of food production as one or the other form of valuable additives. Imparting reduced levels of waste, this
idea will help in reduction of environmental issues and contribute directly towards economic stability. That being said,
the waste biomass management can also be used as a prominent tool in meeting the sustainability, malnutrition, and
food security issues prevailing in world population.
Known for almost 4000 years, saffron is considered as an important cash crop among the medicinal plants in the world
with its application ranging from traditional medicine to culinary practices. The spice has been widely used in dyeing,
in perfumery, as medicinal herb for treating issues like eye problems, as tonic agent, antidepressant, anticarcinogenic,
anti-inflammatory, anti-ageing and as natural pigments in cosmetics. Annual global production of saffron is estimated to
be around 325 tonnes per year [3] and is widely distributed in regions of South-West Asia and the Mediterranean region.
The major production areas are spun around countries like Iran, Italy, Greece, India, Morocco, Azerbaijan, and Spain, with
Iran contributing to 90% of the total production [4]. Production of saffron in the Islamic republic of Iran is concentrated
in the Khorasan province, and the state accounts for 70% of the total global exports. India ranks as a second-largest pro-
ducer, with production matrixes exclusively distributed around Jammu and Kashmir. Saffron consumption in our state
is estimated to be around 20 tonnes per year, which on a global scale is met by other leading producers like Iran, Spain,
and China (Fig. 1) [3]. The spice is obtained from the dried stigma part of the flower. Saffron is known for its volatile and
non-volatile component profile which revolves majorly around 4 main compounds, namely crocin, crocetin, picrocrocin
and safranal. These compounds have a contributory effect on the particular colour, flavour and odour of saffron spice.
The post-harvest operations of the spice are normally performed without any mechanical operation and involves manual
separation of the parts of the flowers from the target stigma. The process of separation is manpower intensive and leads
to the release of large amounts of biomass. It is reported that to approximately 92 g of bio-residues are produced for
every 100 g of flowers during the production of saffron [5]. This emphasizes the possibility or feasibility of involving
these by products in the sustainable production methods. Presently, there is an expanding demand for the development
of natural food components that can be an alternate to synthetic additives. Solutions that satisfy this current need are
considered significant in nature. The practice of using floral bio-residues as a valuable source of bioactive components
for diverse product applications is mounting on a bigger scale. These scales up the feasibility and utilization of higher
percentage by products of saffron processing system effectively. Saffron bio-residues accounts as good sources of protein
(8.17–24.45 g/100 g of dry weight), lipids (2.22–10.73 g/100 g of dry weight), carbohydrates (33.8–64.9 g/100 g of dry
weight), minerals, dietary fractions (14.4–32.3 g) and has a functional profile rich in antioxidants, phenols, anthocyanins
etc. These residual fractions act as a beneficial substitute for stigma part, which is known for their considerable concen-
trations of volatile and non-volatile fractions. With around 80% of floral bio residues formed during the separation of a
particular amount of saffron, these are potential drivers in sustainability conversions. In added to the higher amount of
waste fractions, the nutritional and bioactive profile of these bio fractions which is discussed in detail in coming sections
also contribute towards the significant conversions.

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Fig. 1  Indian production, Export and Import statistics for saffron over the years [3]

The functional potential of these bio residues should be utilized to its maximum to have an extensive and successful
economic, social, and environmental contribution to the system. With an increasing demand for the natural and healthy
additives to the food system as an alternative to the synthetic formulations, these types of resources are of radical impact
on the system. The possibility of their contributions as sustainable option to eradicate the hunger and undernourish-
ment in different populations of underdeveloped and developed nations is inevitable. There is added assistance of these
components towards the demand for food stuffs owing to the growing world population. Environmental factors in the
system involve the reduction in waste accumulation percentages with the successful valorization of the by-products
involved. This contributes to the physical well-being of the ecosystem, which is lacking in recent times. Over and above,
the economical perspective involving extraction and separation of high-valued components from waste fractions comes
as a primary and important endowment while discussing sustainability. Financial contributions involving higher value for
the extracted material from the waste fractions, their applications, and system to produce these high-valued components
generate a lot of employment opportunities along with development in waste utilization. This in turn will render support
to the production systems and personnel like farmers involved in the practice, contributing towards the development
in their standard of living.

2 Saffron plant and flower anatomy

Having its place in Iridaceae family, Crocus sativus L. is a multifunctional and expensive spice element fitting the role of
a culinary ingredient, food flavouring, colouring sources, and a contributory role in perfumes and inks for ages. The sec-
tional separation of each flower consists of six violet coloured tepals, followed by three stamens and red coloured stigma
part partitioned into three threads in general (Refer Fig. 2) [6]. The most important and valuable part of the flower is the
stigma, which is the raw source of saffron. Known as red gold or king of spice, the stigma part of the flower is manually
collected or separated from the flower and is subjected to continuous drying operation. Saffron, known for its culinary
and medicinal properties, also has numerous reported biological activities like antioxidant, antitumor, anticonvulsant,
hypolipidemic, antispasmodic and stomachal stimulant activities [7]. These biological properties of the so-called stigma
part can be contemplated to other parts of the flower, which are discarded as waste. Tepals and stamens are the major
byproducts of the system, and it contributes to a considerable portion of plant material.
The studies around the other parts of the plant or these by products are very limited in number, which unveils their
application potential. The flowers of saffron are reported for their antioxidant activity owing to the varying phenolic
content of the parts attributed to their growth, geographical region of production and other production characteristics.
This valuable natural resource of antioxidants can be employed in food formulations, feeding to the race between syn-
thetic and natural food components. The antifungal, anti-oxidative and cytotoxic nature of other parts like stamen and
perianth give emphasis and validation to the intent of linking the bioactive potential and food formulations in one go [8].

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Fig. 2  Sectional representa-

tion of saffron flower

Bestowing to such an approach will lead to the reduction in losses attributed to the processing sector of the plant and
further increases the economic viability of this crop. For that amalgamation, there should be a detailed understanding
about the composition of these flower by products, their bioactive components and potentiality. This review attempts
to stage an explanation of the product and component profile of saffron bio-residues, along with their biological activity
and common methods of extraction. Given the diverse component and functional profile of these fractions, the applica-
tion is multifaceted, rendering bio-residues as a significant commodity in terms of sustainability.

3 Nutritional compositions of the byproduct fractions

While evaluating the percentage distribution of major by product fractions, it is stated that tepals constitute the most
important bio-fractions of the flower (78 g/100 g) constituting approximately the major portion followed by stamen
and stigma. Both stamen and stigma constitute only less than 25% of the weight fractions. The lightest constituent is
the style, which has less than 1% of the component share. The moisture content values of these parts followed a similar
trend to weight distributions in case of moisture content evaluation. Fresh weight calculations exhibited a higher value
in moisture content of tepals (90.1 ± 0.2 g/100 g) when compared with other regions of stamen and stigma (Table 1).
Contrary to the obtained values, the mineral composition is rich in style with substantially higher percentage of Ca, Fe,
and Na. The author correlates the higher compositional percentage of calcium in style to their role in plant reproduc-
tion [5]. The lowest mineral range were exhibited by stamens and tepals in general. One of the most significant minerals
associated with saffron and its bioproducts is potassium, which has a contributory effect on bone and cardiovascular

Table 1  Nutritional Plant material/parameter Whole flowers Tepals Stigma Stamen Styles
composition of saffron
flower and their different bio Moisture 10.35 ± 0.56 10.76 ± 0.24 3.84 ± 0.08 9.80 ± 0.32 9.86 ± 0.64
residues [5]
Ash 7.39 ± 0.12 6.16 ± 0.05 6.60 ± 0.66 11.43 ± 0.56 8.33 ± 0.07
Protein 10.07 ± 0.24 8.17 ± 0.27 13.63 ± 0.12 24.45 ± 0.32 11.75 ± 0.37
Lipids 3.16 ± 0.49 2.22 ± 0.29 8.76 ± 0.16 10.73 ± 0.38 3.51 ± 0.21
Carbohydrates 61.2 ± 0.2 64.9 ± 0.4 62.0 ± 0.3 33.8 ± 0.8 58.6 ± 1.2
Dietary fibre 14.4 ± 0.1 27.5 ± 0.1 13.8 ± 0.1 32.3 ± 0.1 18.5 ± 0.1
Minerals
P 0.279 ± 0.010 0.232 ± 0.006 0.327 ± 00.03 0.589 ± 0.022 0.296 ± 0.003
Mg 0.113 ± 0.008 0.098 ± 0.010 0.135 ± 0.013 0.309 ± 0.032 0.178 ± 0.012
Ca 0.139 ± 0.002 0.125 ± 0.008 0.107 ± 0.012 0.216 ± 0.004 0.345 ± 0.016
Fe 0.016 ± 0.001 0.028 ± 0.001 0.011 ± 0.001 0.024 ± 0.001 0.035 ± 0.001
K 1.395 ± 0.061 1.353 ± 0.010 1.486 ± 0.037 3.796 ± 0.031 1.816 ± 0.078
Na 0.010 ± 0.001 0.012 ± 0.001 0.010 ± 0.001 0.011 ± 0.001 0.019 ± 0.001

Proximate composition: mean ± SD in g per 100 g of dry weight

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health [9]. There is a higher percentage of K levels attributed to the floral bio residues (1500 mg/100 g dw) compared
to saffron stigmas (1400 mg/100 g dw). The high-protein content of the stamens (24 g/100 g) is attributed to the pollen
grains and can generate up to 12% of their calorific value from proteins. Analogous to this, petals of Rosa micrantha which
is used for culinary operation has a lower protein content and on comparison the ranges are lower to the percentage in
saffron flower residue [10].
Lipids are the least abundant macronutrient present in these bio-fractions, with stamens (10 g/100 g) entailing
the higher percentages. The comparable compositional nature of saffron bio-residues and the whole flowers show
the potential in application of these fractions. Predominant levels of α-linolenic acid were found in floral bio-residues
(6.87–9.28 g/100 g) which is comparable with the saffron stigma portion (5.77–8.50 g/100 g)) emphasizing their role
or contribution towards cardiovascular risk prevention. Higher percentage of carbohydrates were observed in tepals
(64.9/100 g) followed by style and stamen. Bio-residues obtained after stigma removal have higher concentration of
insoluble, soluble, and total DF compared to the whole flower [5]. The soluble DF percentages of bio-residues are equiva-
lent or comparable with the cereal DF percentages, emphasizing their suitability in food applications. Stamens are found
to have the highest insoluble and total DF content among the other parts, whereas the soluble DF is found higher in
tepals. This is in accordance with the results obtained in one of the studies that reported a higher percentage of insoluble
DF in floral by products than in saffron stigmas [9].
Similarly, tepals are noted to be the flower parts consisting of higher concentration of reducing sugars, almost having
a tenfold difference from other parts. Higher glucose percentage were observed in the case of tepals and the lowest
percentages were in stamen. Tepals were reported to have a considerable amount of sucrose, while maltose was present
both in tepal and style fractions of the flower [5]. When it comes to the dietary reference intake (DRI) values, the contri-
butions made by whole flowers and flower bio residues were similar in nature. Dietary reference values of total dietary
fibres were covered in greater proportion by bio residues than whole flowers of saffron. Beyond 70% of the contribution
to DRI of total dietary fibre were made by stamens tailed by the tepals. Higher values of DRIs of calcium were contributed
by bio-residue fractions, rendering the percent of DRIs of calcium by 100 g of bio-residues comparable with that of dairy
products [5]. There is reported proximity and influence of factors like growing conditions, humidity, temperature, and
soil conditions in the development of these proximate elements of the sample.

4 Phytochemistry of the bio‑resource fractions

By product fractions from saffron processing are known for their composition, counting acceptable levels of protein,
carbohydrates, dietary fibres and mineral compounds like calcium, potassium, and sodium. In addition to these proximate
components, there are reports elucidating the presence of different chemical compounds in various parts of the flower.
Flower parts are rich in compounds like carboxylic acids, amino acids, phenolic compounds, flavonoids etc (Table 2).
Carotenoids are the natural pigments responsible for the orange and yellow colour profile of plants and vegetables.
Contributing to the photosynthetic properties, the compounds are also known for their antioxidant potential. Crocin and
crocetin forms the two types of independent carotenoids opulent in saffron stigma along with the occurrence in other
parts of the saffron flower including petals and stamen [11]. The petals were known for the incidence of glycosidic forms
(6 different forms) of crocetin at a concentration of 6.4 mg/g of dried petals. The author also elucidates about the anti-
tumor activity exhibited by the presence of these compounds against many types of tumours even though the mecha-
nism remains indistinct [12]. Terpenoids are a class of compounds which has a large and diverse classification including
monoterpenes, sesquiterpenes etc. contributing to anti-allergic, antioxidant, anticancer, anti-inflammatory properties
[13]. Fresh floral bio residues of saffron are characterized with high levels of monoterpenoids with a concentration of
around 61.63%. Among the different compounds, α-pinene and 1,8-cineole around 13.03% and 20.61% were the most
abundant type in fresh flowers. 1,8-cineole is the major contributory element to the pungent and spice like after-taste of
fresh saffron tepals [8]. The author also elucidates a variation in the number of monoterpenes in dried flower particulates
compared to the fresh ones. Dried flower by products consist of low levels of monoterpenes of around 5.55% along with
high concentration levels of phenylpropanoids (56.99%).
Another class of major compounds found in saffron flower is the flavonoids, which are powerful antioxidants in
nature. Flavanols mainly noted and reported in the floral by products are kaempferol 3-O-sophoroside, quercetin
3-O-sophoroside, isorhamnetin 3-O-glucoside, and kaempferol 3,7,4′-O-triglucoside. Higher concentration of kaempferol
3-O-sophoroside component is accounted in all type of samples (Flower parts devoid of stigma) with values ranging from
25 to 30 mg/gm of dried flower material [15]. Incidence of nineteen different flavanols were detected in saffron tepals

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Table 2  Important Compound Physical structure Target application

phytochemical components
in bi-floral residues, their Crocetin • Antioxidant properties
structure, and some C44H64O24 • Neuroprotective
application [14] • Cosmetic colourant
• Application in dyes

α- pinene • Cosmetic
C10H16 • Food additive
• Flavouring agent

Kaempferol 3-O-sophoroside • Antioxidant, anti-inflammatory and antitu-

C27H30O16 mour activities
• Cosmetic application

1–8-cineole • Flavouring agent

C10H18O • Dehydrating agent
• Used in pharmaceuticals
• Essential oils

Rutin • Anti-inflammatory and Antioxidant nature

C27H30O16 • Stabilizer and colourant
• Cosmetic and personal care products
• Pharmaceutical applications

Epicatechin • Antioxidant properties

C15H14O6 • Used in treatment of diabetes and cancer

Delphinidin • Glazing agent: Food additive

C15H11ClO7 • Cosmetic ingredient
• Antioxidant in nature

Petunidin 3,5-diglucoside • Food additive: colorant

C28H33ClO17 • Antioxidant in nature
• Anti-inflammatory in nature

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with higher concentration of glycosidic forms of kaempferol. The major flavanols portions distributed were kaempferol
glycosides with concentration levels of about 84.0% of total flavanol content, with major distribution of kaempferol
3-O-sophoroside. Similarly, a component profile of 50 flavanols with a concentration of kaempferol-3-O-sophoroside
around 44.21–58.73 mg/g of dried material is observed in the case of total floral bio-residues [16]. In addition to this,
incidence of different other forms of flavanols and flavanols including rutin and epicatechin were reported in the case
of methanolic tepal extracts [6, 17] which shows the diversity of these flower components.
The variance in the composition of the bio-active components in floral bio-residues can be attributed to the difference
in geographical growth conditions, climate, the species, the floral residue parts, and other production parameters. While
comparing the different parts of floral residues, the highest flavonoid content is reported in the case of tepals while the
lowest value of bio-actives concentrated in stamens and styles [18]. The other major phenolic compounds found in these
fractions are anthocyanins, which are pigments responsible for colour and oxidation profile. The major pigment fractions
mentioned in saffron tepal extracts were delphinidin, petunidin and malvidin glycosides [19]. The major anthocyanin
content was attributed to delphinidin-3,7-di-O-glucoside with total concentration ranging around 4804 μg per gram of
dried product. More studies also reported the higher content and concentration of delphinidin-3,5-di-O-glucoside with
a concentration around 2.11–6.37 mg/g of dried material as a major anthocyanin derivative in total floral bio-residues
[16]. Pertaining to the anthocyanin contents, there is observed difference in concentration levels while comparing with
the flavonoid contents of floral residues. The major anthocyanin profile was attributed to delphinidin 3,5-O-diglucoside,
followed by petunidin 3,5-O-diglucoside with their values far less than the flavonoid content [15]. Authors have made
efforts to evaluate the stability of anthocyanin is relation to the temperature and relative humidity and found out that
the most convenient storage conditions are 25 °C and 23% RH, evading anthocyanin loss in saffron whole flowers [20].

5 Extraction methods

To contract any component or material with superior bioactive profile, it is important to have a proper and effective
extraction method along with raw material of recommended quality. Proximate and detailed component evaluation
has ensured the quality of the raw material involved here. Selection of solvents and the techniques involved effects the
efficiency of the extraction process in any system. The prerequisite for maximum extraction of all bioactive components
from saffron floral bio-residue is the selection of a viable process of extraction. Effective ways of combination of water
and other solvents with different techniques like microwave assisted and ultrasound assisted extraction and effective-
ness of these techniques were evaluated and reported in different studies (Table 3).

5.1 Traditional extraction techniques

Maceration is a conventional technique used in the extraction of the bio-actives from floral bio-residues. The only limi-
tation related to the process is the higher extraction times and consumption of organic solvents. The procedure was
reported to be effective in extraction of phenolic compounds including ellagic acid, epicatechin; anthocyanin like cyani-
din 3,5-diglycosides; pelargonidin 3- and 3,5-glycosides, delphinidin 3-glycosides and flavonoid components including
kaempferol and quercetin glycosides. The process of extraction commonly involves solvents like water, methanol, ethanol,
and their acidified forms and sulphur water solution. Among the different combinations of solvents in the maceration
process, ethanolic and methanolic extracts were showing higher efficiency in extraction of bio-actives from saffron
tepals. One amid such studies reported the superiority in extraction of total phenolic content with 80% of methanolic
solutions [17]. Higher quantities of total phenolic contents and higher anthocyanin content were obtained with the
help of the same solution when compared to the effectiveness of separate solvent application or in combination. The
phenolic content of the samples was in the range of 374.8–677.7 mg GAE 100 ­g−1 FW according to the difference in the
solvents involved and their combinations. The authors also report the difference in antioxidant activity of the extracted
material measured with FRAP assays with a variation in extraction method and the solvent utilized. Analogues results
were obtained in the case of juices obtained from the floral by products from saffron. The total phenolic activity was
around 460–743 mg GAE 100 ­g−1 of the floral residues, with correlated effects and deviations in the total antioxidant
activities of the sample [21]. Besides the reported dependence of extraction efficiency with the solvents, there is varia-
tion in extraction results regarding the difference in the raw material involved. The total phenol content was higher in
methanolic extracts of leaves, followed by tepals and corms of saffron flower [22]. There is also a reported difference in

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Table 3  Summary on different extraction technologies involved in treatment of bio residues of saffron
SL no. Extraction method Solvents involved Observations References

1 Maceration Water and ethanol (0, 20, 50 and 80%) Most of the solvent concentrations exhibit almost similar [17]
results, with an exception in the case of 80% ethanol con-
centration. TPC (677.7 mg GAE 100 g ­ −1 of FW) and anthocy-
anin were a bit superior in case of this solvent combination
2 Ohmic heating assisted (OHAE), microwave assisted (MAE), Water Ohmic heating assisted extraction of bioactive from floral bio- [25]
ultrasound assisted extraction (UAE) systems residues exhibited better quality extracts. Conventional hot
treatment extracts were of inferior quality
3 Maceration Ethanol (70%) Kaempferol glycosides were the major components found [22]
in tepals with higher content of phenolics in leaf samples,
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followed by tepals and corms

4 Soxhlet extraction Hexane, dichloromethane and ethanol Maximum extraction yield was obtained using ethanol. Total [23]
phenolic and flavonoid content was higher in case of etha-
nolic extracts, followed by dichloromethane and hexane
extracts
5 Ultrasound assisted extraction Water and ethanol (0, 20, 50 and 80%) Similar results were obtained in case of all the solvent com- [17]
binations. Total antioxidant value was better in case of 50%
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methanol combination for ABTS method

6 Maceration Ethanol (20:1, 80:1) Ethanol concentration of 25.02% was effective in extracting [26]
1609.11 mg/l anthocyanins
7 Maceration Methanol and ethyl acetate Methanolic extracts were better in yield and phenolic content [19]
was higher
8 Microwave mediated extraction procedure Water, glycerine and ethanol (70%) Higher retention rates of total phenols and anthocyanins [15]
were obtained in case of water extracts. Glycerine extracts
showed the lowest value
9 Super critical ­CO2 extraction & conventional solvent extrac- Super critical ­CO2 and water Higher quantitative and qualitative values of different phy- [27]
tion tochemical constituents of floral bio-residues extracted by
SCF
10 Ultrasound assisted extraction & maceration Water and methanol Water extracts of both the techniques exhibited superior [6]
results. Total anthocyanin content of UAE water extracts was
having better yields than all combinations
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efficiency with the state of extracting material, in the sense whether they are fresh or in dried form. All these factors need
to be considered while evaluating the efficiency of this conventional treatment in extraction.
While entailing in study of plant extracts, one of the common and customary procedures that is obvious is the soxhlet
extraction. It is a continuous extraction process that requires less amount of solvent and time in comparison with the
maceration. Reported with higher extraction yields, soxhlet extraction technique was also found to be solvent depend-
ent. The extraction yields were maximum, encircling the presence of ethanol with a maximum number of phenols and
flavonoids in the plant extract. The phenol and flavonoid content of plant extract involving different solvents are ranged
between 45 and 84 mg GAE/g and 32–68 µg QE/g [23]. The antioxidant, antimicrobial, and other phytochemical activity of
the extract also followed a correlation with the concentration, as the extracts with higher concentration showing higher
potential. Similar to these results, the concentration dependency of the Crocus sativus L extracts in their antimicrobial
efficacy was reported in another related work [24].

5.2 Modern or green extraction techniques

As the conventional techniques involve the utilization of higher percentage of chemicals, there is always a pursuit for
better extraction technologies. The increased consumer demand for healthy, safe, and nutritional potential food and
their additives has also furthered this quest. These factors led to the introduction of green extraction technologies like
ultrasound assisted extraction, microwave mediated extractions which ensured efficacy, extraction quality, lower extrac-
tion time and less or no use of chemicals. Authors have reported the effectiveness and superiority of microwave mediated
extraction procedure using water as a solvent [15]. The total phenol content of water extracts was higher than the ethanol
extracts of floral bio-residues, followed by lower value of glycerine extracts. The author proposes the microwave mediated
extraction method in water as a better extraction method to recover the maximum number of qualitative fractions of
flavonoids and anthocyanins. This is in correlation with the effectiveness and quality superiority of water extracted floral
bio residue samples and saffron stigma samples as anti-inflammatory, anti-oxidative and enzyme inhibitory agents [11].
Another operative technique involved in the extraction of bioactive components from floral residues is the ultrasound
assisted extraction (UAE). While comparing maceration and UAE for acquiring high-quality extracts, the amount of total
phenols in water UAE extracts were comparable with macerated results in less timings. Meaning, the performance rate
of UAE technique was better than the conventional treatment, ensuring the better quality of the extracted components
owing to less time. Considering the trend followed in extraction of anthocyanin content, a similar upper hand of the
UAE water extracts was visible. There is a percentage variation from the average yield percentage exhibited by the water
extracts over all the solvent combinations in anthocyanin extraction. This ensures the suitability and viability of solvent
free additives to the food components as the customer demand solicits.
Along with the solvent combination, there is a major influence of treatment technique in successive extraction
of bio-active components [25]. The superiority of green techniques like ohmic heating assisted extraction (OHAE),
microwave assisted extraction (MAE) and ultrasound assisted extraction (UAE) systems over conventional hot water
treatment were observed in the study with higher values of phenolics, flavonoids, and anthocyanin perceptible in
these treatments. Among the three green extraction procedures, both UAE and MAE systems showed almost similar
results regarding mentioned parameters. Ohmic heating assisted extraction of bioactive from floral bio-residues
exhibited better quality extracts with superior component profile of the sample. This again emphasizes the affinity
of extracting solvents and procedures towards the quantitative and qualitative efficacy of the process. Ahmadian
and Niazmand, [27] have reported the effectiveness of super critical fluid extraction, a novel extraction technique in
extracting the biomolecules of industrial importance from saffron bio-residues. The super critical extraction process
resulted in higher amounts of polyphenols, flavonoids, and elevated antioxidant activity while compared with con-
ventional extraction method. The shorter time of the SC-CO2 process has also helped in maximizing the characteristics
of antioxidants compounds. The unique dissolving properties of SC-CO2 contributes to the efficiency of the process.
[28]. With the extraction procedure, process parameters are also reported to have an impact on the quantitative
and qualitative profile of bioactive components. Solvent ration was found to be one of the important factors influ-
encing the extraction yield of the microwave assisted extraction of bioactive from saffron by products. Microwave
assisted extraction was a rapid and efficient technique in extracting 101 mg anthocyanins/g at a temperature of
48 °C, solvent to sample ratio of 77.5 ml/g and extraction time of 9.3 min [29]. The antioxidant activity, total phenols
concentration and yield are found to be affected by the treatment time and temperature along with the solvent ratio.
Optimal microwave extraction conditions for the bioactive material in saffron by products were reported to be an
amalgamation of low temperature (25 °C), moderate extraction time (5 min) and ethanol (100%) as a solvent [30].

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Extracts displayed maximum values of total phenolic content as 126.20 ± 2.99 mg gallic acid equivalent (GAE)/g dry
matter, total flavonoid content as 8.05 ± 0.11 mg catechin equivalent (CE)/g dry matter, and antioxidant activity as
6219 ± 246 μmol trolox equivalent antioxidant capacity (TEAC)/g dry matter. Constructive changes were induced in
the incorporation of microwave as a pretreatment in conventional extraction of bioactive, which includes a reported
reduction in the extraction times of up to 12 times. A microwave pretreatment at temperature of 65ºC and solvent
ratio of 0.30 g/mL was found to be effective in boosting the antioxidant capacity, yield composition and polyphenol
richness by 15% [31]. For every process, it is important to understand the characteristics, influencing parameters and
other factors which directly or indirectly affect the process and the extract quality.

6 Application of floral saffron bio‑residues

Accounting as one of the expensive elements among the spice fractions, the feasibility of production and marketing will
be augmented with the effective utilization of their bio residues. The production of floral bio-residues is inevitable in this
case, as stigma accounted as spice is the primary resource. The theory of effective production, handling, processing, and
packaging will not be applicable to a higher extent in these types of product fractions. Exploring the component profile
and effective utilization of the respective fraction is the only sustainable option put forward. Owing to the high-valued
ingredient profile, phytochemistry and nutritional benefits of these waste bio fractions, their application is diversified
into different fields which are explained in detail.

6.1 Food industry

Relevance in food-based applications is always related to the nutritional, functional and physicochemical properties
of the plant material and its constituent profile. The stigma part of the flower has been involved in households and
industrial applications extensively for a very long period of time [32]. Saffron bio-residues owing to their nutrient and
functional profile can be effectually engrossed in the practice of food enrichment. There are reported literature studies
illustrating about the involvement of saffron floral bio-residues as a sustainable vegan ingredient in wheat and spelt
breads with an aim of improving their functionality and nutritional quality [33]. Compared to the control sample, mineral
and nutritional percentages exhibited an elevated and positive effect in concentration of minerals, inulin, organic acids
etc. along with higher values of phenolic content. The antioxidant potential was also on the higher side, with a direct
correlation with the added percentages of floral bio-residues. A reported 25–30% increase in dietary fibre content was
observed in traditional bread recipes, altering the textural properties of the samples. Envisaging the possibility of novel
bakery products for vegans and vegetarians, the addition of saffron floral residues can be a suitable alternative without
compromising on the beneficial effect of food intake on human health. Further, the extracts from the bio-fractions had
a contributory effect towards the shelf-life extension of shrimp products [34]. Shrimp treated with water, ethanolic and
methanolic extracts of tepal had the lowest lipid oxidation, melanosis and bacterial count causative in superior sensory
properties up to 9 days.
Analogues to these products, there is productive improvement in yogurt incorporated with different concentra-
tion of saffron and its bio-residues by including them in traditional recipe formulation [35]. Encapsulated in alginate
beads, the extracts were found to be potent functional ingredients in enhancing the bioactive and functional profile of
yogurt samples. Without getting in the way of the fermentation process, microbiological and physicochemical properties,
enriched samples were presented with elevated levels of organic acids and sugars with high total phenol percentages
and antioxidant properties. A saffron-based probiotic beverage can be proposed as an apt and healthy substitute for
caloric and carbonated beverages. The higher phenolic, antioxidant profile and stability profile of these beverages with
four strains of lactic acid bacteria shows that saffron-based beverage is a good medium for the growth and production
of a functional beverage [36]. Being a non-dairy beverage, it can also be recommended for lactose intolerant patients.
However, there is only a limited amount of research carried out on the utilization of these floral components of saffron.
With a potentiality of these low-cost components, there should be sustainable approaches encouraging the production
of functional, nutritional and novel formulations.

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6.2 Nutraceutical application of bio‑residues

The bioactive component range of the byproducts justifies their application in modern pharmacological formulations
and could be the reason for their involvement in traditional medicine [8]. The antimicrobial activity of the component
fractions shows a dependency on the type of the extract and also on the plant part from where the extract is made.
Ethereal extracts of saffron flower stamens exhibit a notable antimicrobial activity both invitro and in food matrix
against some common strains of food-borne bacterial species [24]. These results can be allied directly to the com-
ponent profile including considerable amounts of flavonoids, polyphenols, polyunsaturated fatty acids like linolenic
acids and terpenes. Comparable report on the antimicrobial activity of saffron petal extracts against major pathogens
was studied and reported [37]. One among the most sensitive strains to petal extracts was Salmonella typhimurium,
and the resistant one were E. coli and Staphylococcus aureus. It is reported that among the different parts, the high-
est antibacterial properties were displayed in petals, and style whereas stamens were more effective as antifungal
agents [38]. The aqua and methanolic extracts showed better antibacterial activity than ethanolic extracts of petals,
which elaborates the formerly explained correlation among the extracts [37].
Tepal fractions can be considered as a new and potential source of kaempferol and glycosides components with
a notable antioxidative property [41]. The total phenolic and flavonoid content of the tepal extracts is on a higher
side compared to the stigma extracts, which shows direct correlation with the antioxidant properties of the samples
[39]. The in vitro antioxidant activity of plant fractions assessed by oxygen radical absorbance capacity (ORAC) assay
and 2,2-Diphenyl-1-picrylhydrazyl (DPPH) scavenging assay confirms this elucidation. Both the values display higher
percentage of activity in tepal extract, but the former assay elucidates the possibility of twofold higher activity in
tepal fractions [39]. The antioxidant profile of these plant fractions is allied with the diverse phytochemical profile,
which links them towards other practical pharmacological applications. The anticancer properties presented by the
petal extracts were explained in detail [49]. The saffron petal extracts were highly sensitive towards HepG2 cells and
displayed significant cytotoxicity against cancer cells. The author correlates the activity of petal extracts with the
presence of crocin and crocetin in the extracts, even though the mechanism remains unclear. The cancerous cells
in the body are presumed to be targeted by these phytochemical fractions, causing sensitive of cells. The flower
extracts showed an excellent antitumour activity against the carcinoma cell line, with an ­LC50 value of 75 showing
the effectiveness [45]. Analogous, the petal extracts of saffron also displayed a ­IC50 value of 10.8 mg/ml compared
to that of stigma fractions having a value of 5.3 mg/ml against tumour cells [50].
Further, the flower constituents are known for their effectiveness as anti-inflammatory and anti-depressant markers
(Table 4). The petal extracts of saffron has known to contribute in the treatment of mild to moderate depression and
being a cheaper option, the use of petal is economically feasible as well. In comparison with fluoxetine, a major drug
in anti-depressive disorders, the petal extracts had a similar effect to the former drug in the treatment of patients
[51]. Tepal and stigma fractions displayed a comparable effect in the treatment of mild to medium depression issues
in a randomized trial for a period of 6 weeks [52]. This is in coordination with the study described [53] regarding
the causative stimulus of the compound crocin in managing and in treatment of psychiatric disorders and mental
health. As discussed, bio fractions are also known sources of these phytochemical compounds as the saffron (stigma
portion). Crocus sativus L. stigma and petal extracts have shown significant antinociceptive and anti-inflammatory
effects in mice. Research indicates that the stigma extract displayed maximum analgesic activity and edema inhibi-
tion, followed by the petal extract. Additionally, the ethanolic extract from Crocus sativus L. petals exhibited anti-
inflammatory activity by reducing nitric oxide production and IL-6 release in macrophages, suggesting its efficacy
in inflammation modulation [44]. Furthermore, both ethanolic and aqueous extracts of Crocus sativus were found
helpful in moderating neuropathic pain by suppressing proinflammatory cytokines, enhancing antioxidant activity,
and affecting apoptotic pathways in chronic constriction injury animal models [42]. These findings collectively high-
light the therapeutic potential of Crocus sativus L. extracts in managing pain and inflammation in preclinical models.

6.3 Other applications

Crocus sativus L. floral bio-residues have shown significant potential in skin protection due to their antioxidant, anti-
inflammatory, and tissue-protective properties [8]. These bio-residues, particularly saffron tepals, contain various
bioactive compounds that contribute to their therapeutic benefits, making them a valuable ingredient in cosmetology

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Table 4  Review on the pharmacological potential of floral bio-fractions of saffron

Vol:.(1234567890)
SL no. Saffron by products Potential effect Observations References

1 Petals Antimicrobial activity Aqueous and methanolic extracts were better against the inhibition of bacterial populations. The [37]
most sensitive fractions of bacterial populations were reported to be E. coli and Staphylococcus
aureus
2 Tepals Antimicrobial & antioxidant activity Extracts were effective in interfering the biofilm formation of two methicillin resistant staphylococcal [39]
reference strains, inhibiting the bacterial growth in total. The antioxidant properties were found to
be higher than the compared stigma extracts
3 Petals Anti-inflammatory & reproductive health Extracts had a role in reducing the no of cystic follicles and thereby decreasing the probability of ovar- [40]
ian cyst formation. Anti-inflammatory properties of the petal extract were evident by the decrease
in the inflammatory markers such as IL6, IL18, CRP, IL1β, and TNFα
4 Stamens Antimicrobial activity Diethyl ether extracts exhibited notable levels of antimicrobial activity against strains of S. aureus and [24]
E. coli
5 Tepals Antimicrobial and Antioxidant activity Extracts exhibited antimicrobial properties against six different bacterial species, and the minimum [41]
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quantity was around 1.5 mg of tepal extract. Phenolic content ranged between 5.5 to 39 mg Caffeic
Acid Equivalent/g source material with notable antioxidant activity
6 Stigma and Petals Analgesic, and antidepressant activity Stigma extracts exhibited the highest analgesic activity and had a contribution towards edema inhibi- [42]
tion, followed by petal extracts. Both the extracts can be categorized with depressant efficacy with
immobility times of less than 76 s
7 Petals and Stamens Antimicrobial activity Antibacterial activity of methanolic extracts of petals were more prominent against Salmonella [43]
enterica, Staphylococcus aureus and Shigella dysenteriae. The stamen extracts were effective against
(2025) 6:123

B. cereus & E. coli

8 Petals Anti-inflammatory and anti-osteoclasto- Extracts controls the inflammation by decreasing nitric oxide and IL-6 release and are demonstrated [44]
genic activity to exhibit an anti-osteoclastogenic effect by inhibiting the responsible genes and osteoclast forma-
tion
9 Flower Antimicrobial activity Extracts were found to be effective in controlling the antimicrobial populations of different patho- [45]
genic microbes, with inhibition zone diameter ranging from 15 to 20 mm and having a mean index
of 18.7 mm
10 Petal Antibacterial activity It exhibited efficacy against gram positive bacterial populations like C. perfringes, C. botulinum etc. and [46]
the inhibition zone diameter extended between 13 and 18 mm
11 Flower waste Antioxidant and anti-microbial activity Extracts from bio-residues of saffron has antimicrobial effect against fungi like Botrytis cinerea and [47]
bacteria like Staphylococcus aureus
12 Petal Antioxidant and cardiovascular effects Extract displayed in enhancing the cardiovascular condition owing to its intrinsic activities on smooth [12]
muscles. The antioxidant potential of the extract is correlated with the presence of crocin and
kaempferol which has a role in the reduction of reactive oxygen formation
13 Petal Anti-inflammatory activity Invitro and in-vivo experiments showed the efficacy of petal extracts in controlling the inflammation. [48]
In comparison with diclofenac, there is similar efficacy exhibited by tepal extracts and was in the
range of 63% in vitro and 71% in-vivo models
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and perfumery [54]. Studies have demonstrated that saffron administration can enhance skin flap viability, reduce
oxidative stress, and decrease apoptotic cell death after skin flap surgery, ultimately improving skin health and heal-
ing processes [55]. Additionally, the ethanolic extract from Crocus sativus L. petals has exhibited anti-inflammatory
activity and anti-osteoclastogenic effects, suggesting its potential in preventing bone disruption and supporting
overall skin health through its protective and healing properties [44]. Ethanolic and aqueous extracts of tepal were
effective in moderating the oxidative stress induced by UV and hydrogen peroxide exposures. Tepal extracts are
known for subduing the cytotoxicity nature of sodium dodecyl sulphate which causes skin and eye irritation [15].
Furthermore, the saffron flower extract has been successfully used as a natural colorant for sustainable textile dyeing,
showing good stability under specific conditions and producing colours ranging from brown to green with excellent
colour fastness on wool fabric [56]. The authors also recommend optimized conditions for better dyeing performance
of wool with dye concentration of 6% for direct and 2% for mordant dyeing at a temperature of 90º, pH value of 3
and the time of 60 min. This comprehensive approach addresses waste management and highlights the multifaceted
benefits of utilizing saffron flower waste for dye extraction and antioxidant effectiveness.

7 Conclusion

Food waste or by product processing to generate high-valued and useful products for functional and industrial applica-
tions is gaining scientific as well as economic interest. Several studies explain saffron by products as a beneficial resource
of some significant components like polyphenols, flavonoids etc. In contemporary scenario, the successful utilization
of floral bio residues from the saffron processing system will entice more attention among multiple facets. The known
potential of the bio-residues as in proximate composition and bio active potential is contributory to the development of
the field. The functionality and superior component profile is highly dependent on the processing technologies and the
parameter considerations. Despite the potential, there is a lack of involvement of different technologies in the extrac-
tion system and the application studies of the extracted components. Incorporation of suitable green and innovative
technologies will be obliging in augmenting the process efficacy and ensures the quality of the extracted component.
The research process is majorly concentrated on the traditional methods of extraction and if there is an involvement of
certain other procedures the system lacks the investigation on effect of parameters on the yield. Research work focussing
on understanding the importance of parameters on the extraction yield and quality is lacking, and this incomprehension
will affect the utilization of these bio fractions. Imperative necessity is there in understanding how these novel techniques
are positively influencing the final quality and quantity of the extracts. Added to that, there are fewer studies involved
in checking the efficacy of extracted bio-actives in different food formulations. Understanding about the factors and
application diversities will give an idea about the improvements needed in the system for achieving acceptable levels
of extraction efficacy and application feasibility.

Author contributions A.S.M: Contributions to the conception or design of the work; performed the literature search and data analysis, and
drafting of the work; A.P. Revised it critically for important intellectual content, and reviewed the version to be published; S.S.R. Revised it
critically for important intellectual content and reviewed the version to be published.

Funding Open access funding provided by Vellore Institute of Technology. No funding was received to assist with the preparation of this
manuscript.

Data availability No datasets were generated or analysed during the current study.

Declarations
Competing interests The authors declare no competing interests.

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adapta-
tion, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source,
provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article
are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in
the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will
need to obtain permission directly from the copyright holder. To view a copy of this licence, visit [Link]

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