INTRODUCTION
According to the International Diabetes Federation
report of 2017, approximately 425 million adults
between 20 and 79 years old suffered from diabetes
worldwide. By 2045, this number will escalate to
629 million. In 2017, India reported 72 946 400 cases
of diabetes [1]. Type II diabetes patients showed
higher cancer risks, especially in the colorectal area.
Association between these two diseases may result
from shared cellular and molecular pathways. Genomewide
association studies also linked diabetes-associated
genes (e.g., TCF7L2) to colorectal cancer [2, 3].
Globally, colorectal cancer is the fourth most commonly
diagnosed type of cancer. The past five years have seen
3.2 million prevalence rates. It means that 1.3 million
new colorectal cancer cases are registered every year [4].
According to Ayurvedic studies, food (Ahara in
Hindi) is the sustainer of life, which helps maintain
good health and protects human body from diseases [5].
284
Dhakshayani GM et al. Foods and Raw Materials. 2022;10(2):283–294
Herbs and spices are indispensable parts of human
diet. Since ancient times, herbs and spices have played
a vital role in the lifestyle of people. Not only do they
add flavor to food, but they also possess valuable
preservative and medicinal properties because the biomolecules
in some plants maintain and promote human
health.
In the past few decades, natural products have
become more popular as an alternative therapy against
various diseases because conventional medicine often
cause unwanted side effects. As a result, modern science
also started exploring the medicinal properties of
spices [6, 7].
Coriander (Coriandrum sativum), sometimes called
the herb of happiness, is the most well-known culinary
spice worldwide and an age-old traditional medicine.
C. sativum contains a wide range of phytochemical
elements, which makes it a promising functional
food that protects from all kinds of lifestyle-related
diseases. Indeed, coriander is known for its antioxidant,
anticancer, neuroprotective, anticonvulsant, migrainerelieving,
hypolipidemic, hypoglycemic, hypotensive,
antimicrobial, anxiolytic, analgesic, and antiinflammatory
activities [8].
Mature coriander leaves have medicinal
properties, but new scientific data demonstrate that
coriander microgreens contain higher amounts of
such phytonutrients as β-carotene, ascorbic acid,
α-tocopherol, and phylloquinone, as well as minerals,
e.g., Ca, Mg, Fe, Mn, Zn, Se, and Mo. They also have
lower nitrate content than mature leaves [9, 10].
As a novel functional food, microgreens are tender
and immature leafy greens with developed cotyledons
and with or without partially emerged pair of the first
true leaves [10]. They are harvested for consumption
within 10 to 20 days of seedling emergence and are
larger than sprouts but younger than baby greens [11].
They give vivid color, soft texture, and multifarious
quality to the main dish, thus enhancing its aesthetic
appeal [12, 13]. Microgreens are a highly perishable
food with a very short shelf life of three to five days at
ambient temperature [14]. Microgreens can be easily
grown at home, in containers on a terrace, or in kitchen
gardens with minimal sunlight. In the present study,
the microgreens were evaluated in vitro for antioxidant,
antidiabetic, and anticancer properties, which were
compared with those of mature leaves.
STUDY OBJECTS AND METHODS
Sample growth and preparation. Coriander
(Coriandrum sativum) microgreens were grown under
ambient conditions using vermicompost enriched soil.
A 50-g sample of coriander seeds (Chennai, India) was
sown at an even depth of one inch (2.5 cm) in soil-filled
plastic pots. After germination, the pots were hydrated
thrice a day and exposed to ambient light. Coriander
microgreens were harvested after seven or eight days
when they were three inches (7.5 cm) tall. The cotyledon
stems were cut with sterile scissors as close to the soil
surface as possible. Coriander mature leaves were
grown under the same conditions as microgreens and
harvested after 60 days. The roots and defected parts
were removed, and the edible stems and leaves were
cleaned from soil particles.
Species identification. The species were identified
with the help of the faculty of Plant Biology and Plant
Biotechnology, Women’s Christian College, Chennai.
Preparation of extract. Mature leaves and microgreens
were washed three or four times with tap water
and then rinsed twice with de-ionized water. After
that, they were shade-dried at room temperature under
constant observation to avoid any contamination. After
drying, the leafy samples were crushed in an electric
grinder. The powdered samples were stored for further
use. Extraction was done by aqueous and ethanolic
methods.
Aqueous extraction. Powdered mature leaves (10 g)
and powdered microgreens (10 g) were put in separate
conical flasks with 100 mL of de-ionized water. The
samples were kept in a water bath at 90°C for 1 h and
cooled at room temperature. Then, the extract was
filtered with Whatman filter paper. The filtrate was
condensed in a hot plate at 50°C and stored at 4°C.
Ethanolic extraction. Powdered mature leaves
(10 g) and powdered microgreens (10 g) were soaked
separately in 100 mL of ethanol for 72 h. The
supernatant was filtered with Whatman filter paper. The
filtrate was condensed in a hot plate at 50°C.
Phytochemical analysis. Qualitative phytochemical
screening. The crude ethanolic and aqueous
extracts of C. sativum microgreens and mature leaves
were subjected to a qualitative phytochemical analysis.
They were tested using standard procedures for various
classes of active phytoconstituents, such as alkaloids,
terpenoids, glycosides, steroids, saponins, tannins,
flavonoids, phenols, carbohydrates, and proteins [15–21].
Quantitative phytochemical analysis. Estimation
of total phenols. Total phenolic compounds in the
coriander samples were quantified by using a slightly
modified the Folin-Ciocalteu reagent method [22].
During the procedure, 100 μL of extracts were mixed
with 900 μL of methanol and 1 mL of the Folin-
Ciocalteu reagent (diluted with distilled water as 1:10).
After 5 min, 1 mL of 20% (w/v) Na2CO3 solution was
added. The reaction was incubated in the dark for 30
min. A UV-Vis spectrophotometer measured the optical
density at 765 nm. The total phenolic content was
expressed as (mg/g of sample) gallic acid equivalent.
Estimation of total flavonoids. The aluminum
chloride reagent method with slight modifications
was used to define the total flavonoid content in the
C. sativum samples [23]. Each extract (500 μL) was
mixed with 500 μL of methanol and 500 μL of 5% (w/v)
sodium nitrite solution followed by adding 500 μL of
10% (w/v) aluminum chloride solution. After a 5-min
incubation, 1 mL of 1M NaOH solution was added. By
adding distilled water, the total volume was brought up
285
Dhakshayani GM et al. Foods and Raw Materials. 2022;10(2):283–294
to 5 mL. Absorbances were measured at 510 nm, and
the results were expressed as (mg/g of sample) quercetin
equivalent.
Estimation of steroids. According to the procedure
described in [24], 1 mL of each extract was put in a
10-mL volumetric flask. 4 N sulphuric acid (2 mL) and
0.5% iron (III) chloride (2 mL) were added, followed
by a 0.5% potassium hexacyanoferrate (III) solution
(0.5 mL). The mix was heated at 70 ± 20°C in a water
bath for 30 min with occasional shaking. The total
volume was diluted to the mark with distilled water.
The optical density was measured at 780 nm against the
reagent blank. The results were expressed as (mg/g of
sample) cholesterol equivalent.
Estimation of total carbohydrates. The total
carbohydrate content was measured by the Hedge and
Hofreiter method [25]. According to the procedure,
0.5 mL of each extract was put in a separate test tube.
The volume was brought up to 1 mL with distilled
water. After that, 4 mL of anthrone reagent was added
in each tube and mixed thoroughly. D-glucose was
used as standard. Blank was taken as distilled H2O and
anthrone. The reaction mix was heated in a boiling
water bath for 8 min and cooled. The absorbance of
the green color solution was tested at 630 nm using a
UV-Vis spectrophotometer. The carbohydrate content
of the plant extract was calculated from the calibration
curve of glucose, and the results were expressed as
(mg/g of sample) glucose equivalent.
Estimation of proteins (Bradford colorimetric assay).
The Bradford protein assay described in [26] quantified
the total protein content in the C. sativum samples.
According to the procedure, 0.5 mL of each extract was
put in a test tube and brought up to 1 mL with distilled
water. After that, 2 mL of Bradford’s reagent was added
in each tube and mixed thoroughly. Bovine serum
albumin served as standard. Blank was taken as distilled
water and Bradford’s reagent. The absorbance of the pale
blue color solution was tested at 595 nm. The unknown
concentration of amino acids/protein in the coriander
samples was illustrated as a graph.
Estimation of ascorbic acid. The ascorbic acid
content in the fresh samples were estimated using the 2,
6-dichlorophenol indophenol (DCPIP) titration method
according to the procedure previously described by Rao
and Deshpande [27]. According to the procedure, 5 mL
of the ascorbic acid working standard was pipetted into
a 100 mL conical flask together with 5 mL of 0.625%
oxalic acid and titrated against the dye solution (V1).
The endpoint was the appearance of a transient pink
color that persisted for a few minutes. After that, 5 mL
of each test sample was similarly titrated against the
dye solution. The ascorbic acid content, mg/100 g, was
determined using the following formula:
(1)
where 500 is the amount of standard ascorbic acid taken
for titration, μg; V1 is the volume of dye consumed by
500 μg of standard ascorbic acid; V2 is the volume of dye
consumed by 5 mL of each test sample; 25 is the total
volume of extract; 100 is the ascorbic acid content per
100 g of sample; 5 is the weight of fresh sample taken
for extraction; 5 is the volume of test sample taken for
titration.
Estimation of chlorophylls and carotenoids using
acetone. During this procedure, 1 g of finely cut fresh
leaves was homogenized with 80% acetone. The mass
was then centrifuged at 5000 rpm for 5 min. After
the supernatant was transferred, the procedure was
repeated until the residue contained no trace of green
color. The final volume was brought up to 100 mL in the
volumetric flask with 80% acetone. The optical density
of the extracted solution was measured at 480, 510, 645,
and 663 nm. From these readings, concentrations of
chlorophylls and carotenoid pigment were determined
by using the following formulas given in Table 1.
Gas chromatography–mass spectrometry
(GC/MS). The aqueous extracts of C. sativum
microgreens and mature leaves underwent a GC/MS
analysis by using Agilent technologies 6890 N JEOL
GC Mate II GC-MS model. The samples were injected
into an HP-5 column (30 m×0.25 mm i.d with 0.25 μm
film thickness). During the gas chromatography, helium
served as the carrier gas, the flow rate was 1 mL/min,
and the injector operated at 200°C. The column oven
temperature was programmed as 50–250°C at a rate of
10°C/min injection mode. The list of mass spectrometry
Positive control optical Growth inhibition density Sample Positive control optical density
−
= % of -amylase enzyme inhibition Sample Control 100
Sample
−
α = ×
% of Fe3 reduction Sample Control 100
Sample
+ −
= ×
% of DPPH radical inhibition Control Sample 100
Control
−
= ×
% of DPPH radical inhibition Control Sample 100
Control
−
= ×
2
1
Amount of ascorbic content 500 25 100
5 5
× × ×
=
× ×
V
V
% Cell viability =100 − Percent growth inhibition
% of -amylase enzyme inhibition Sample Control Sample
−
α = ×
Table 1 Formulas for chlorophyll-a, chlorophyll-b, total chlorophyll, and carotenoid estimation [28–30]
Chlorophyll-a, mg/g tissue
Chlorophyll-b, mg/g tissue
Total chlorophyll (TC), mg/g tissue
Carotenoid, mg/g tissue
where A is the absorbance at a specific wavelength (480, 510, 645, and 663 nm); V is the final volume of chlorophyll extract; W is the fresh weight
of tissue extracted
( ) ( ) 645 663 20.2 8.02
1000
 − × × 
 
 
A A V W
( ) ( ) 480 510 7.6 1.49
1000
 − × × 
 
 
A A V W
( ) ( ) 663 645 12.7 2.69
1000
 − × × 
 
 
A A V W
( ) ( ) 645 663 22.9 4.68
1000
 − × × 
 
 
A A V W
( ) ( ) 645 663 20.2 8.02
1000
 − × × 
 
 
A A V W
( ) ( ) 480 510 7.6 1.49
1000
 − × × 
 
 
A A V W
( ) ( ) 663 645 12.7 2.69
1000
 − × × 
 
 
A A V W
( ) ( ) 480 510 7.6 1.49
1000
 − × × 
 
 
A A V W
( ) ( ) 645 663 20.2 8.02
1000
 − × × 
 
 
A A V W
( ) ( ) 480 510 7.6 1.49
1000
 − × × 
 
 
A A V W
( ) ( ) 663 645 12.7 2.69
1000
 − × × 
 
 
A A V W
( ) ( ) 645 663 22.9 4.68
1000
 − × × 
 
 
A A V W
( ) ( ) 645 663 20.2 8.02
1000
 − × × 
 
 
A A V W
( ) ( ) 480 510 7.6 1.49
1000
 − × × 
 
 
A A V W
( ) ( ) 663 645 12.7 2.69
1000
 − × × 
 
 
A A V W
( ) ( ) 480 510 7.6 1.49
1000
 − × × 
 
 
A A V W
1000
 
 
( ) ( ) 480 510 7.6 1.49
1000
 − × × 
 
 
A A V W
( ) ( ) 663 645 12.7 2.69
1000
 − × × 
 
 
A A V W
( ) ( ) 645 663 22.9 4.68
1000
 − × × 
 
 
A A V W
( ) ( ) 645 663 20.2 8.02
1000
 − × × 
 
 
A A V W
( ) ( ) 480 510 7.6 1.49
1000
 − × × 
 
 
A A V W
( ) ( ) 663 645 12.7 2.69
1000
 − × × 
 
 
A A V W
( ) ( ) 480 510 7.6 1.49
1000
 − × × 
 
 
A A V W
( ) ( ) 480 510 7.6 1.49
1000
 − × × 
 
 
A A V W
( ) ( ) 663 645 12.7 2.69
1000
 − × × 
 
 
A A V W
( ) ( ) 645 663 22.9 4.68
1000
 − × × 
 
 
A A V W
( ) ( ) 645 663 20.2 8.02
1000
 − × × 
 
 
A A V W
( ) ( ) 480 510 7.6 1.49
1000
 − × × 
 
 
A A V W
( ) ( ) 663 645 12.7 2.69
1000
 − × × 
 
 
A A V W
( ) ( ) 480 510 7.6 1.49
1000
 − × × 
 
 
A A V W
286
Dhakshayani GM et al. Foods and Raw Materials. 2022;10(2):283–294
conditions included: ionization voltage – 70 eV; ion
source temperature – 250°C; interface temperature –
250°C; mass range – 50–600 mass units. The results
were compared using the spectrum of the known
components stored in the National Institute Standard
and Technology (NIST) library database [31].
In vitro antioxidant assays. DPPH radical
scavenging assay. The antioxidant activity of
the extracts was measured based on the stable (2,
2-diphenyl-1-picryl-hydrazyl-hydrate) DPPH free radical
scavenging method [32]. Various concentrations (50, 100,
150, 200, 250, and 300 μg/mL) of C. sativum extracts
(1 mL) were mixed with 0.1 mM of DPPH solution
(1 mL) in methanol. The reaction was carried out in
triplicate, and the decrease in absorbance was measured
at 517 nm after 30 min in the dark using a UV-Vis
spectrophotometer. Ascorbic acid served as the standard
reference, while methanol (1 mL) with DPPH (1m L)
solution served as control. The percentage of inhibition
was calculated as follows:
The procedure made it possible to determine the
sample concentration required to inhibit 50% of the
DPPH free radical (IC50).
Ferric (Fe3+) reducing antioxidant power assay
(FRAP). The reducing power of the extracts was
determined by the Fe3+ reduction method with slight
modification [33]. In brief, 1 mL of C. sativum extracts
at different concentrations (50, 100, 150, 200, 250, and
300 μg/mL) were taken in 1 mL of phosphate buffer
(0.2 M, pH 6.6) in a test tube. After that, 1 mL of
potassium ferricyanide [K3Fe(CN)6] (1% w/v) was added.
After 30 min of incubation at 50°C in a water bath,
1 mL of trichloroacetic acid (10 % w/v) was added to
each mix. Then, 1 mL of fresh FeCl3 (0.1% w/v) solution
was poured in, and the absorbance was measured at 700
nm in a UV-Vis spectrophotometer. The experiment
was replicated in three independent assays. Ascorbic
acid was used as the standard reference. The reducing
concentration (RC50) of sample required to reduce the
free radicals (Fe3+) by 50 % was calculated to interpret
the FRAP results.
The percentage of reduction was calculated as
follows:
In vitro antidiabetic activity. α-amylase
enzyme inhibition assay. The α-amylase enzyme
inhibition assay relied on the starch-iodine test [34].
The coriander extracts at various concentrations (50,
100, 150, 200, 250, and 300 μg/mL) were added to
α-amylase enzyme (10 μL). The α-amylase enzyme
had been prepared in 0.02 M sodium phosphate buffer
(pH 6.9 containing 6 mM sodium chloride). The
procedure was followed by 10 min of incubation at
37°C. After pre-incubation, 500 μL of 1% soluble starch
was added to each reaction and incubated at 37°C for
60 min.
To stop the enzymatic reaction, 1 N HCl (100 μL)
was added and followed by 200 μL of iodine reagent
(5 mM I2 and 5 mM KI). The color change was
registered, and the optical density was tested at
595 nm. Acarbose was used as the standard reference.
The control reaction representing 100% enzyme activity
contained no plant extract.
The experiment was carried out in triplicate. A darkblue
color indicated the presence of starch; a yellow
color indicated the absence of starch; a brownish color
indicated partially degraded starch in the reaction mix.
In the presence of inhibitors, the starch added to the
enzyme assay mix did not degrade and gave a darkblue
color complex. No color complex developed in
the absence of the inhibitor, indicating that starch was
completely hydrolyzed by α-amylase. The IC50 value was
calculated as follows:
Cytotoxicity assay on colon cell lines. The
conventional MTT reduction assay was used to measure
the cell viability [35]. HT 29 Colon cells were obtained
from the National Centre for Cell Science (Pune). The
culturing was performed on the medium developed by
the Roswell Park Memorial Institute (RPMI). It included
10% fetal bovine serum (FBS), gentamycin (100 μg/mL),
penicillin/streptomycin (250 U/mL), and amphotericin
B (1 mg/mL). All cell cultures were maintained
at 37°C in a humidified atmosphere of 5% CO2. Cells
grew to confluence for 24 h before use.
As described in [36], we plated HT 29 cells (5×103/
well) in 96-well plates for 24 h in 200 μL of the RPMI
medium with 10% fetal bovine serum. After the culture
supernatant was removed, the RPMI samples with
various concentrations (0.001–100 μg/mL) of aqueous
C. sativum extracts were added and incubated for
48 h. After the treatment, cells were incubated with
MTT (10 μL, 5 mg/mL) at 37°C for 4 h and then with
dimethyl sulfoxide at room temperature for 1 h. The
plates were tested at 595 nm on a scanning multi-well
spectrophotometer. All experiments were performed in
duplicates [36].
The effect of the extracts on growth inhibition of
HT-29 colon cancer cell line line, %, was calculated
using the following formula:
(2)
(4)
(5)
(3)
287
Dhakshayani GM et al. Foods and Raw Materials. 2022;10(2):283–294
phenol (116.78 mg GAE/g) in comparison to that of
mature leaves (72.23 mg GAE/g). In general, both
extracts of microgreens had more total flavonoids,
steroids, carbohydrates, and proteins than both extracts
of mature leaves. Table 3 illustrates the contents of
ascorbic acid, chlorophyll, and carotenoid.
Gas chromatography–mass spectrometry (GC/MS).
The GC/MS method revealed various bioactive
constituents in the aqueous extracts of coriander
microgreens and mature leaves. The analysis showed
peaks at different locations on the chromatogram. In
Figs. 1 and 2, the X-axis represents the retention time,
while the Y-axis represents the relative abundance. The
GC/MS analysis of a crude extract of microgreens
showed nine major peaks. The crude extract of mature
leaves eluted seven major peaks. Tables 4 and 5 illustrate
a comparative analysis of the mass spectra of the
constituents with the NIST library data.
In vitro antioxidant assays. DPPH radical
scavenging assay. The scavenging capacity of the
aqueous and ethanol extracts of both coriander
microgreens and mature leaves on DPPH free radicals
was expressed as inhibition (%) (Tables 6 and 7). The
IC50 was inhibition concentration at 50%: the lowest
IC50 indicated the strongest ability of the extracts to act
as DPPH radical scavengers. The aqueous and ethanol
extracts of mature leaves showed the lowest IC50, which
were 44.64 and 186.74 μg/mL, respectively. As for the
aqueous and ethanol extracts of microgreens, they were
90.09 and 293.54 μg/mL, respectively. Compared to the
reference standard ascorbic acid inhibition percentage
(Fig. 3), the test samples required higher concentration
to inhibit DPPH free radical. Thus, the test samples of
microgreens and mature leaves showed dose-dependent
scavenging activity.
Ferric (Fe3+) reducing antioxidant power assay.
For Fe3+ reducing activity, the ascorbic acid was used as
Table 2 Phytochemical content of aqueous and ethanol extracts of Coriandrum sativum microgreens and mature leaves
Phytochemicals
Aqueous extract Ethanol extract
Microgreens Mature leaves Microgreens Mature leaves
Phenols, mg GAE/g 98.25 ± 0.27* 107.26 ± 0.29* 116.78 ± 0.28* 72.23 ± 0.28*
Flavonoids, mg QE/g 119.43 ± 0.36* 18.58 ± 0.38* 29.15 ± 0.26* 13.61 ± 0.37*
Steroids, mg CE/g 140.34 ± 0.57* 101.77 ± 0.28* 50.41 ± 0.52* 33.58 ± 0.38*
Carbohydrates, mg GE/g) 457.65 ± 1.6* 398.38 ± 2.3* 169.73 ± 1.50* 124.35 ± 1.04*
Proteins, mg/g 156.41 ± 0.38* 117.80 ± 0.31* 101.40 ± 0.37* 75.36 ± 0.35*
Each value is expressed as mean ± standard deviation (n = 3) and statistically significant at *P < 0.05
Table 3 Ascorbic acid, chlorophyll, and carotenoid contents in Coriandrum sativum microgreens and mature leaves
Samples Phytonutrients
Ascorbic acid,
mg/100 g W
Chlorophyll-a,
mg/g W
Chlorophyll-b,
mg/g W
Total chlorophyll,
mg/g W
Carotenoid,
mg/g W
Microgreens 18.56 ± 0.45* 0.04 ± 0.01* 0.07 ± 0.01* 0.04 ± 0.01* 0.13 ± 0.01*
Mature leaves 77.68 ± 0.37* 0.27 ± 0.04* 0.33 ± 0.03* 0.33 ± 0.03* 0.31 ± 0.04*
Each value is expressed as mean ± standard deviation (n = 3) and statistically significant at *P < 0.05
From the above growth inhibition, (%) percentage of
cell viability was derived using the following formula:
Statistical analysis. The phytochemical, antioxidant,
and antidiabetic assays were carried out in triplicates,
while the anticarcinogenic analysis was carried out
in duplicates. The results obtained were expressed as
mean ± SD. The statistical analysis was calculated by
one-way ANOVA and Student’s t-test using Microsoft
excel. All statistical significance was accepted at
P < 0.05.
RESULTS AND DISCUSSION
Phytochemical analysis. Qualitative phytochemical
analysis. The qualitative phytochemical analysis of the
aqueous and ethanolic extracts of coriander microgreens
and mature leaves revealed such phytochemicals as
alkaloids, terpenoids, steroids, tannins, flavonoids,
phenols, carbohydrates, and proteins. Saponins were
absent in both aqueous and ethanol extracts of
microgreens and mature leaves. However, glycosides
were present in the aqueous extract of microgreens and
mature leaves, as well as in the ethanol extract of mature
leaves. However, they were absent in the ethanol extract
of microgreens.
Quantitative phytochemical analysis. Tables 2
shows the quantitative phytochemical mean values
of both aqueous and ethanol extracts of Coriandrum
sativum microgreens and mature leaves.
According to Table 2, the aqueous extract of
microgreens showed a lower total phenol content
(98.25 mg GAE/g) than that of mature leaves
(107.26 mg GAE/g). However, the ethanol extract of
microgreens had significantly (P < 0.05) higher total
(6)
288
Dhakshayani GM et al. Foods and Raw Materials. 2022;10(2):283–294
Figure 2 Bioactive constituents identified in coriander mature leaves
20.58
21.60 23.43
12.77
15.05
15.78
17.03
19.00
200 300 400 500 600 700 800 900 1000 1100 1200 1300
10 15 20 25 30
7000000
Scan
Min
20.58
21.60 23.43
12.77
15.05
15.78
17.03
19.00
14000000
21000000
28000000
35000000
42000000
49000000
56000000
63000000
Figure 1 Bioactive constituents identified in Coriandrum sativum microgreens
14.90
16.00
18.55
18.28
20.0521.15
23.67
25.50
200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500
14.90
16.00
18.55
18.28
20.0521.15
23.67
25.50
5 10 15 20 25 30 35
1000000
2000000
3000000
4000000
5000000
6000000
7000000
8000000
9000000
10000000
11000000
12000000
13000000
Scan
Min
Figure 3 DPPH standard curve of ascorbic acid
86.75 88.68 91.74 93.67
96.16 97.88
y = 11.338x + 33.919
R² = 0,4849
0
20
40
60
80
100
120
0 5 25 30
Percentage inhibition
10 15 20
Concentration, μg/mL
Ascorbic acid Linear (Ascorbic acid)
86.75 88.68 91.74 93.67
96.16 97.88
y = 11.338x + 33.919
R² = 0,4849
0
20
40
60
80
100
120
0 5 25 30
Percentage inhibition
10 15 20
Concentration, μg/mL
Ascorbic acid Linear (Ascorbic acid)
standard. Figure 4 illustrates the standard curve; Tables
8 and 9 show the reducing power of test samples.
The aqueous extract of mature leaves showed
a slight increase in Fe3+ reduction compared to
that of microgreens. The RC50 (50% reducing
concentration) of microgreens and mature
leaves in the aqueous extracts were 234.87 and
167.25 μg/mL, respectively. Interestingly, the ethanol
extract of microgreens exhibited a greater ferric ion
reducing power (31.66% at 300 μg/mL concentration)
than that of mature leaves (18.77% at 300 μg/mL
concentration). The ethanol extracts were unable
to reduce the free radicals by RC50. The causes may
be in some other chemical constituents that
compete for reduction by Fe3+ and do not permit Fe3+
289
Dhakshayani GM et al. Foods and Raw Materials. 2022;10(2):283–294
Table 4 GC/MS analysis of bioactive compounds in Coriandrum sativum microgreens
RT Name Structure Mol.wt
g/mol &
Mol. Formula
Biological Activity
14.9 Benzene, (1-methylenebutyl)- 146
C11H14
n.d.a.
16 10-Methylundecanoic acid methyl ester 214
C13H26O2
n.d.a.
16.55 (7-Phenyl-1H-imidazo[4,5-d] pyridazin-4-
yl)-hydrazine
226
C11H10N6
Anticancer, antidiabetic,
antiviral, antiosteoporotic, antiinflammatory,
antiparasitic,
antihypertensive
17.48 Phenol, 2,6-bis(1,1-dimethylethyl)-4-ethyl-
(Phenol)
234
C16H26O
Antioxidant, cytotoxicity,
antidiabetic
18.28 Propenamide,2-acetamido-3-Phenyl-N-(3-
hydroxypropyl)-(amide)
262
C14H18N2O3
Antioxidant
20.05 8-carbetoxy-1-methyl-1,4,5,6,7,8-
hexahydropyrrolo[2,3-b]azepin-4-one-3-
carboxylic acid
280
C13H16N2O5
n.d.a.
21.15 5-Phenyl-5,6,7,8-tetrahydro-[1]
benzothieno[2,3-d]pyrimidine-2,4-diamine
296
C16H16N4S
Antioxidant, antitumor,
anticancer, antidiabetic,
antimicrobial, antiviral, antiinflammatory
23.67 Z-13-Octadecen-1-yl acetate
(Essential oil)
310
C20H38O2
Antioxidant, anti-inflammatory
25.5 But-2-endiamide,N,N’-bis[4-methoxyphenyl]- 326
C18H18N2O4
n.d.a.
n.d.a. – no data available
Table 5 GC/MS analysis of bioactive compounds in Coriandrum sativum mature leaves
RT Name Structure Mol.wt
g/mol &
Mol. Formula
Biological Activity
12.77 2,4-bis[1,1-dimethylethyl]-phenol
(Phenolic compound)
206.00
C14H22O
Antioxidant, antibacterial,
anti-inflammatory
15.05 1-Cyclopentenylphenylmethane 158.00
C12H14
n.d.a.
15.78 7-Dodecen-6-one
(Terpenoid)
182.00
C12H22O
Antioxidant, antibacterial, anti-fungal,
anti-malarial
17.03 E, E-6,8-Tridecadien-2-ol, acetate
(Essential oil)
238.00
C15H26O2
Antimicrobial
19 2-Hexadecenoic acid,
2,3-dimethyl-, methyl ester, (E)-
(Unsaturated fatty acid ester)
296
C19H36O2
Antioxidant, antidiabetic, antitumor,
antibacterial, anti-inflammatory, anthelmintic,
immunostimulant, lipoxygenase inhibitor
20.58 3-Hydroxypregn-5-en20-one
(Steroid)
316
C21H32O2
Anti-proliferative
23.43 Methoxyaceticacid, octadecyl
ester
342
C12 H42 O3
n.d.a.
n.d.a. – no data available
290
Dhakshayani GM et al. Foods and Raw Materials. 2022;10(2):283–294
Table 6 DPPH radical scavenging activity of Coriandrum sativum microgreens and mature leaves (aqueous extract)
Extract concentration,
μg/mL
Inhibition, % Test samples (IC50 μg/mL) Standard ascorbic acid (IC50 μg/mL)
Microgreens Mature leaves Microgreens Mature leaves
50 49.51 ± 0.49* 56.22 ± 0.47*
90.09 ± 0.45 44.64 ± 0.46 2.88 ± 0.37
100 55.41 ± 0.45* 66.66 ± 0.48*
150 59.17 ± 0.30 * 70.00 ± 0.50*
200 65.44 ± 0.47* 75.55 ± 0.45*
250 76.44 ± 0.47* 77.75 ± 0.46*
300 81.17 ± 0.30* 84.63 ± 0.41*
Each value is expressed as mean ± standard deviation (n = 3) and statistically significant at * P < 0.05
Table 7 DPPH radical scavenging activity of Coriandrum sativum microgreens and mature leaves (ethanol extract)
Extract concentration,
μg/mL
Inhibition, % Test samples (IC50 μg/mL) Standard ascorbic acid (IC50 μg/mL)
Microgreens Mature leaves Microgreens Mature leaves
50 7.35 ± 0.39* 14.20 ± 0.30*
293.54 ±0.36 186.74 ± 0.43 2.88 ± 0.37
100 22.34 ± 0.41* 31.88 ± 0.34*
150 37.90 ± 0.36* 39.48 ± 0.45*
200 42.79 ± 0.26* 53.42 ± 0.36*
250 45.56 ± 0.32* 65.44 ± 0.39*
300 51.16 ± 0.30* 75.57 ± 0.32*
Each value is expressed as mean ± standard deviation (n = 3) and statistically significant at * P < 0.05
Figure 5 Standard curve of acarbose
0
33.3
46.71
63.24
68.65
76.36
82.18
y = 12.664x + 2.2629
R² = 0.9011
0
10
20
30
40
50
60
70
80
90
100
0 10 20 30 40 50 60 Percentage inhibition
Concentration, μg/mL
Acarbose Linear (Acarbose)
17.81
36.94
41.47 44.18
45.69
51.59
y = 7.7775x + 2.8443
R² = 0.8334
0
10
20
30
40
50
60
70
0 5 10 15 20 25 30
Percentage reduction
Concentration, μg/mL
Ascorbic acid Linear (Ascorbic acid)
86.75 88.68 91.74 93.67
96.16 97.88
y = 11.338x + 33.919
R² = 0,4849
0
20
40
60
80
100
120
0 5 25 30
Percentage inhibition
10 15 20
Concentration, μg/mL
Ascorbic acid Linear (Ascorbic acid)
Figure 4 FRAP standard curve of ascorbic acid
86.75 88.68 91.74 93.67
96.16 97.88
y = 11.338x + 33.919
R² = 0,4849
0
20
40
60
80
100
120
0 5 25 30
Percentage inhibition
10 15 20
Concentration, μg/mL
Ascorbic acid Linear (Ascorbic acid)
Cytotoxicity assay on colon cell lines. In
the present study, the antioxidant activities of the
aqueous extracts were compared to those of
the ethanolic extracts. The aqueous extract of
microgreens and mature leaves were examined for
potential anticancer activity against the human colon
HT-29 carcinoma cell line by using the MTT assay.
The tests were performed in duplicate. The absorbance
values were registered in the ELISA reader at 595 nm
once purple color developed after 24 h of incubation.
The mean was calculated for two trials (Table 12).
to donate an electron. The RC50 value for standard
ascorbic acid was 29.1 μg/mL.
In vitro antidiabetic activity. α-amylase enzyme
inhibition assay. Tables 10 and 11 show the inhibitory
activity of test samples on the α-amylase enzyme. The
aqueous and ethanol extracts of microgreens exhibited
50% of inhibition on α-amylase enzyme at 222.22
and 84.25 μg/mL. The results were lower than those
of mature leaves IC50 values, which were 228.31 and
206.82 μg/mL, respectively. The standard reference drug
acarbose (Fig. 5) showed α-amylase inhibitory activity
with an IC50 valueof 23.71 μg/mL.
291
Dhakshayani GM et al. Foods and Raw Materials. 2022;10(2):283–294
As the concentration of the test samples
increased, the corresponding absorbance value
decreased (P < 0.05). The MTT assay showed that
the microgreen sample increased the percentage
inhibition and consequently decreased the cell
viability to 49.08% with the lowest IC50 value
of 98.34 μg/mL. Mature leaves showed the least
percentage inhibition and reduced viable cells
to 59.53% with an IC50 value of 123.54 μg/mL
(Figs. 6 and 7). Doxorubicin was used as the reference
standard. Figure 8 demonstrates the standard
curve of percent cell viability, which showed
a cytotoxicity activity with an IC50 value of
11.75 μg/mL.
Table 8 Ferric reducing antioxidant power activity of Coriandrum sativum microgreens and mature leaves (aqueous extract)
Extract concentration,
μg/mL
Reduction, % Test samples (RC50 μg/mL) Standard ascorbic acid (RC50 μg/mL)
Microgreens Mature leaves Microgreens Mature leaves
50 22.64 ± 0.21* 19.23 ± 0.25*
234.87 ± 0.33 167.25 ± 0.36 29.10 ± 0.36
100 25.50 ± 0.35* 33.79 ± 0.31*
150 35.36 ± 0.31* 44.36 ± 0.29*
200 48.42 ± 0.33* 59.79 ± 0.32*
250 53.22 ± 0.23* 60.60 ± 0.36*
300 60.31 ± 0.27* 64.64 ± 0.35*
Each value is expressed as mean ± standard deviation (n = 3) and statistically significant at * P < 0.05
Table 10 α-amylase enzyme inhibition activity of Coriandrum sativum microgreens and mature leaves (aqueous extract)
Extract concentration,
μg/mL
Inhibition, % Test samples (IC50 μg/mL) Standard ascorbic acid (IC50 μg/mL)
Microgreens Mature leaves Microgreens Mature leaves
50 15.66 ± 0.42* 9.57 ± 0.30*
222.22 ± 0.37 228.31 ± 0.31 23.70 ± 0.34
100 29.83 ± 0.23* 37.82 ± 0.31*
150 39.27 ± 0.37* 39.16 ± 0.29*
200 46.45 ± 0.37* 45.61 ± 0.44*
250 56.25 ± 0.30* 54.75 ± 0.23*
300 87.31 ± 0.36* 79.55 ± 0.36*
Each value is expressed as mean ± standard deviation (n = 3) and statistically significant at * P < 0.05
Table 9 Ferric reducing antioxidant power activity of Coriandrum sativum microgreens and mature leaves (ethanol extract)
Extract concentration,
μg/mL
Reduction, % Test samples (RC50 μg/mL) Standard ascorbic acid (RC50 μg/mL)
Microgreens Mature leaves Microgreens Mature leaves
50 11.27 ± 0.27* 9.52 ± 0.35*
Nil Nil 29.10 ± 0.36
100 15.41 ± 0.37* 10.59 ± 0.30*
150 15.80 ± 0.32* 13.83 ± 0.25*
200 19.32 ± 0.32* 14.85 ± 0.24*
250 20.04 ± 0.40* 16.50 ± 0.36*
300 31.66 ± 0.38* 18.77 ± 0.28*
Each value is expressed as mean ± standard deviation (n = 3) and statistically significant at * P < 0.05
Table 11 α-amylase enzyme inhibition activity of Coriandrum sativum microgreens and mature leaves (ethanol extract)
Extract concentration,
μg/mL
Inhibition, % Test samples (IC50 μg/mL) Standard ascorbic acid (IC50 μg/mL)
Microgreens Mature leaves Microgreens Mature leaves
50 45.70 ± 0.35* 11.84 ± 0.24*
84.25 ± 0.25 206.82 ± 0.35 23.70 ± 0.34
100 59.35 ± 0.39* 33.44 ± 0.37*
150 65.10 ± 0.31* 40.45 ± 0.36*
200 68.82 ± 0.21* 46.58 ± 0.36*
250 69.46 ± 0.34* 60.44 ± 0.39*
300 69.73 ± 0.25* 61.13 ± 0.27*
Each value is expressed as mean ± standard deviation (n = 3) and statistically significant at * P < 0.05
292
Dhakshayani GM et al. Foods and Raw Materials. 2022;10(2):283–294
Figure 6 Effect of aqueous extracts of Coriandrum sativum
microgreens and mature leaves on growth inhibition of HT-29
colon cell line
also had a higher α-amylase enzyme inhibitory property
and a greater anticarcinogenic effecton colon cancer
cell line. Therefore, C. sativum microgreens proved
to be amore effective antioxidant, antidiabetic, and
anticarcinogenic agent than mature leaves. Coriander
microgreens can be as good as mature coriander leaves
for the daily diet of a disease-free community.
CONTRIBUTION
The authors are equally involved in writing
the manuscript and are equally responsible for
plagiarism.
CONFLICT OF INTEREST
The authors have declared no conflict of interests
regarding the publication of this manuscript.

1. International Diabetes Federation [Internet]. [cited 2022 Jan 10]. Available from: https://www.idf.org/aboutdiabetes/what-is-diabetes/facts-figures.html

2. González N, Prieto I, del Puerto-Nevado L, Portal-Nuñez, S, Ardura JA, Corton M, et al. 2017 update on the relationship between diabetes and colorectal cancer: Epidemiology, potential molecular mechanisms and therapeutic implications. Oncotarget. 2017;8(11):18456-18485. https://doi.org/10.18632/oncotarget.14472

3. Suh S, Kim K-W. Diabetes and cancer: is diabetes causally related to cancer? Diabetes and Metabolism Journal. 2011;35(3)193-198. https://doi.org/10.4093/dmj.2011.35.3.193

4. Meyer B, Are C. Current status and future directions in colorectal cancer. Indian Journal of Surgical Oncology. 2018;9(4):440-441. https://doi.org/10.1007/s13193-017-0711-9

5. Nishteswar K. Ayurvedic concept of food and nutrition. Journal of Nutrition & Food Sciences. 2017;6(4). https://doi.org/10.4172/2155-9600.1000530

6. De Souza EL, Stamford TLM, De Oliveira Lima E, Tarajano VN, Barbosa Filho JM. Antimicrobial effectiveness of spices: An approach for use in food conservation system. Brazilian Archives Biology Technology. 2005;48(4):549-558. https://doi.org/10.1590/s1516-89132005000500007

7. Sachan AKR, Kumar S, Kumari K, Singh D. Medicinal uses of spices used in our traditional culture: World wide. Journal of Medicinal Plants Studies. 2018;6(3):116-122.

8. Prachayasittikul V, Prachayasittikul S, Ruchirawat S, Prachayasittikul V. Coriander (Coriandrum sativum): A promising functional food toward the well-being. Food Research International. 2018;105:305-323. https://doi.org/10.1016/j.foodres.2017.11.019

9. Pinto E, Almeida AA, Aguiar AA, Ferreira IMPLVO. Comparison between the mineral profile and nitrate content of microgreens and mature lettuces. Journal of Food Composition and Analysis. 2015;37:38-43. https://doi.org/10.1016/j.jfca.2014.06.018

10. Xiao Z, Lester GE, Luo Y, Wang Q. Assessment of vitamin and carotenoid concentrations of emerging food products: Edible microgreens. Journal of Agricultural and Food Chemistry. 2012;60(31):7644-7651. https://doi.org/10.1021/jf300459b

11. Ebert AW, Wu TH, Yang RY. Amaranth sprouts and microgreens-A homestead vegetable production option to enhance food and nutrition security in the rural-urban continuum. SEAVEG2014: families, farms, food: Proceedings of the regional symposium on sustaining small-scale vegetable production and marketing systems for food and nutrition security; 2014; Bangkok. Taiwan: AVRDC; 2015. p. 233-244.

12. Kou L, Yang T, Luo Y, Liu X, Huang L, Codling E. Pre-harvest calcium application increases biomass and delays senescence of broccoli microgreens. Postharvest Biology and Technology. 2014;87:70-78. https://doi.org/10.1016/j.postharvbio.2013.08.004

13. Mir SA, Shah MA, Mir MM. Microgreens: Production, shelf life and bioactive components. Critical Reviews in Food Science and Nutrition. 2017;57(12):2730-2736. https://doi.org/10.1080/10408398.2016.1144557

14. Chandra D, Kim JG, Kim YP. Changes in microbial population and quality of microgreens treated with different sanitizers and packaging films. Horticulture Environment and Biotechnology. 2012;53(1):32-40. https://doi.org/10.1007/s13580-012-0075-6

15. Banu SK, Cathrine L. General techniques involved in phytochemical analysis. International Journal of Advanced Research in Chemical Science. 2015;2(4):25-32.

16. Finar IL. Stereo chemistry and the chemistry of natural products. Vol. 2. Longman; 1986. 518 p.

17. Hindi NKK, Al-Charrakh AH, Naher HS, Abbas AS. Study of chemical analysis of Iraqi propolis and active component of proplis, Iraq. Journal of Science. 2015;15(11):1095-1103.

18. Kokate CK. Practical pharmacognosy. 4th edition. New Delhi: Vallabh Prakashan; 1994. 186 p.

19. Shrivastava DK. Phytochemical analysis of a miracle herb Coriander sativum. Indian Journal of Scientific Research. 2017;13(2):09-14.

20. Singh V, Kumar R. Study of phytochemical analysis and antioxidant activity of Allium sativum of Bundelkhand region. International Journal of Life Sciences Scientific Research. 2017;3(6):1451-1458.

21. Wagner H, Bladt S. Plant drug analysis. A thin layer chromatography atlas. Heidelberg: Springer Berlin; 1996. 384 p. https://doi.org/10.1007/978-3-642-00574-9

22. Singleton VL, Orthofer R, Lamuela-Raventós RM. Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methods in Enzymology. 1999;299:152-178. https://doi.org/10.1016/S0076-6879(99)99017-1

23. Ahmed D, Fatima K, Saeed R. Analysis of phenolic and flavonoid contents, and the anti-oxidative potential and lipid peroxidation inhibitory activity of methanolic extract of Carissa opaca roots and its fractions in different solvents. Antioxidants. 2014;3(4):671-683. https://doi.org/10.3390/antiox3040671

24. Kharate MS, Pandhure NB. Quantitative estimation of phytochemical and antimicrobial properties of crude leaf extract, Mimusops elengil L. International journal for research in applied science & engineering technology. 2017;5(12):2699-2707.

25. Hedge JE, Hofreiter BT. Carbohydrate chemistry. 17th edition. New York: Academic Press; 1962.

26. Mahavi MO, Nilesh PS. Quantitative estimation of amino acids and carbohydrates in the root exudates of Salvadora persica. International Journal of Biology Research. 2016;1(3):4-10.

27. Rao BS, Deshpande V. Experimental biochemistry: A student companion. Royal Tunbridge Wells: Anshan; 2006. 302 p.

28. Aron DI. Copper enzymes in isolated chloroplasts phenoloxidase in Beta vulgaris. Plant physiology. 1949;24(1):1-15. https://doi.org/10.1104/pp.24.1.1

29. Duxbury AC, Yentach CS. Plankton pigment monograph. Journal of Marine Research. 1956;15:190-191.

30. Maclachalam S, Zalik S. Plastid structure, chlorophyll concentration and free amino acid composition of a chlorophyll mutant of barely. Canadian Science Publishing. 1963;4:1053-1062.

31. Ravisankar N, Sivaraj C, Seeni S, Joseph J, Raaman N. GC-MS Analysis and anticancer activity of methanol extract of leaves of Hypericum hookerianum Wight & Arn. International Journal of Pharmacy and Pharmaceutical Sciences. 2014;6(5):515-519.

32. Khalaf NA, Shakya AK, Al-othman A, El-agbar Z, Farah H. Antioxidant activity of some common plant. Turkish Journal of Biology. 2008;32(1):51-55.

33. Oyaizu M. Studies on products of browning reaction: antioxidative activities of products of browning reaction prepared from glucosamine. Japanese Journal of Nutrition. 1986;44(4):307-315. https://doi.org/10.5264/eiyogakuzashi.44.307

34. Gayathri S, Sivaraj C, Sangeetha ST, Arumugam P, Manimaran A. Evaluation of antioxidant, antibacterial, alpha amylase enzyme inhibition activities and GC-MS analysis of leaves extract of Gymnema sylvestre L. Journal of Pharmacognosy and Phytochemistry. 2018;7(6):2326-2333.

35. Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. Journal of Immunological Methods. 1983;65(1-2):55-63. https://doi.org/10.1016/0022-1759(83)90303-4

36. Vardhini SP, Sivaraj C, Arumugam P, Himanshu R, Kumaran T, Baskar M. Antioxidant, anticancer, antibacterial activities and GCMS analysis of aqueous extract of pulps of Aegle marmelos (L.) Correa. The Journal of Phytopharmacology. 2018;7(1):72-78. https://doi.org/10.31254/phyto.2018.7115