International Journal of Environmental Science and Technology

https://doi.org/10.1007/s13762-022-04121-y

ORIGINAL PAPER

Bioactivity of medicinal plant extracts from Peganum harmala

and Cichorium intybus against Tribolium castaneum Herbst
(Coleoptera: Tenebrionidae)
F. Ahmad1 · A. Javaid1 · T. Zaheer1 · Q. Saeed2 · Z. Ali1 · A. Shabbir1 · S. Naseem1

Received: 14 May 2021 / Revised: 12 January 2022 / Accepted: 13 March 2022

© The Author(s) under exclusive licence to Iranian Society of Environmentalists (IRSEN) and Science and Research Branch, Islamic Azad University 2022

Abstract
The increase in grains production would hardly have any significance, unless protected from the post-harvest losses. One of
the insect pests that cause severe qualitative and quantitative losses to stored grains is Tribolium castaneum. For decades,
insecticides are being used to control insect pests, but they pose significant environmental hazards. Here, we are testing
plant extracts as alternative pesticides for grain protection and the environmental safety. The ethanol and methanol extracts
of Peganum harmala and Cichorium intybus were used with five different concentrations to test their insecticidal activity
against T. castaneum. The extracts of both plants have shown pronounced effects on pest mortality where 40 × ­103 ppm
concentration of both the ethanol and methanol extracts resulted into above 80% larval mortality. These results are time
dependent and the ­LD50 values decreased with the exposure time and the least concentration of 6.9 and 6.1 × ­103 ppm for
methanol and ethanol extracts of P. harmala and C. intybus, respectively, were obtained on 15th days after treatments. The
analysed results showed that the extracts of both P. harmala and C. intybus have a good insecticidal activity on T. castaneum
and can be used to further drug discovery in the quest of novel insecticidal compounds.

Keywords Botanical compounds · Contact toxicity · Harmal · Chicory · Larval mortality

Introduction Tenebrionidae) is one of the most destructive pests of stored

The nature of global grain markets dictates that grain com-
modities need to be stored when supply exceeds the demand.
Hence, they are stored, after harvesting, for prolonged
periods during which many insect pests attack the grains,
causing depreciation in grain quality. It is estimated that
5–10% of world's grain production is lost due to insect pests
(Khaliq et al. 2014). T. castaneum (Herbst) (Coleoptera:
Editorial responsibility: Ta Yeong Wu.
F. Ahmad and A. Javaid have contributed equally to this work.
* S. Naseem
saadia.naseem@comsats.edu.pk
1
Department of Biosciences, COMSATS University
Islamabad (CUI), Park Road Chak Shehzad, Islamabad,
Pakistan
2 lent, antifeedant, sterilization, and toxic effects in insects
Department of Entomology, Faculty of Agricultural Sciences
and Technology, Bahauddin Zakariya University, Multan,
Pakistan
commodities and is of great concern. Indiscriminate use of
synthetic insecticides is associated with many health and
environmental risks, including soil and water contamination,
food contaminations along with detrimental effects on ben-
eficial and non-target organisms (Bughio and Wilkins 2004).
Use of fumigants for the management of stored grain pests
has been the sole remedy for long, due to their non-persistent
nature and efficacy against insects in concealed environ-
ments. However, due to the environmental hazards some of
these fumigants like methyl bromide, are being phased out
(UNEP 2003) while the other, phosphine, is rendered less
effective due to the rapid development of resistance (Kherad-
pir 2014). Therefore, alternate control methods that are safer
for the environment, economical, and convenient to apply
are mandatory to be developed.
Botanical extracts can be a suitable candidate for new
insecticidal compound screening (Cox 2004; Kubo 2006) as
plant extracts contain compounds that show ovicidal, repel-
(Isman 2006; Nawrot and Harmatha 2012). Many of such
medicinal plants have been reported to have potent toxicity

13
Vol.:(0123456789)

against stored product pests (Arnason et al. 1990; Arroyo
1995; Carlini and Grossi-de-Sá 2002; Isman 2006; Jacob-
son 1982; Jilani et al. 1988). Screening of native plant spe-
cies has led to the discovery of many promising compounds
(Javaid et al. 2021; Rates 2001) such as pyrethrum from
the Chrysanthemum cinerariaefolium (Secoy and Smith
1983), azadirachtins (Jilani and Amir 1987), nicotinic acids
from Nicotiana tabacum, piertrins and rotenones from Lon-
chocarpus species. Here in this study, we are evaluating two
of the native plant species, i.e. P. harmala (Zygophyllaceae)
and C. intybus (Asteraceae) for their insecticidal potential
against T. castaneum.
P. harmala is a highly branched deciduous or short-lived
perennial herb with narrow leaves organized alternately on
fleshy, bright green rigid stems (Niroumand et al. 2015). It
is widely distributed in North Africa, the Middle East, Paki-
stan, India, and Iran (Asghari and Lockwood 2002; Ehsan-
pour and Saadat 2002; Yousefi et al. 2009). The enriched
chemical composition of P. harmala L. makes it a suitable
choice to be used as a botanical insecticide (Kartal et al.
2003). The seed extracts of P. harmala contain harmaline,
harmine, harmalol, harmol, and tetrahydroharmine (Fig. 1)
(Bukhari et al. 2008; Pitre and Srivastava 1987; Sharaf
et al. 1997). These extracts have shown promising results
against locusts, Schistocerca gregaria (Forskal) (Orthop-
tera: Acrididae) (Abbassi et al. 2003) and Aedes aegypti (L.)
larvae (Sathiyamoorthy et al. 1997). Moreover, P. harmala
alkaloids have been tested for their phytopharmacological,
therapeutic and toxicological aspects for treating diseases
in higher vertebrates (Arshad et al. 2008; Fan et al. 1997;
Shahverdi et al. 2005; Tahraoui et al. 2007; Zaker et al.
2007). Not only insects, botanical extracts of P. harmala
has shown antifungal potentials as well for treating citrus
diseases (Ameziane et al. 2007).
P. harmala (Zygophyllaceae) most commonly, is used for
the treatment of irregular fevers. Its seeds contain different
chemical components which include harmolol, harmine,
Fig. 1  Major chemical constituents of P. harmala, a Harmane, b
Harmine, c Tetrahydroharmine and d Harmalol (Siddiqui et al. 1987;
Sobhani et al. 2002)
13
International Journal of Environmental Science and Technology
hermaline, and peganine. Antimicrobial, (Al-Shamma et al.
1981; Arshad et al. 2008; Harsh and Nag 1984) antifungal,
(El Bahri and Chemli 1991) antiprotozoal (Arshad et al.
2008) and anticancer (Al-Allaf and Rashan 1998; Lamchouri
et al. 1999) effects of P. harmala extract have been reported.
C. intybus is another short-term perennial herb distributed
in Asia and Europe and has good metabolic characteristics.
High concentrations of quercetin-3-galactose (Kirtikar and
Basu 2012), inulin (Pushparaj et al. 2007), saponins, and
tannins (Shad et al. 2013), caffeic acid, cichoric acid, pectin,
lactucin, cholin, and cichoriin are the major constituents in
the root tissues (Fig. 2). These compounds have been known
for their antifeedant, toxic, or repellent effects against com-
mon insect pest species of economic importance (Nandago-
pal and Kumari 2007).
In this study, we evaluate the efficacy of ethanol and
methanol extracts of P. harmala and C. intybus seeds against
immature stages of T. castaneum. The estimation of lethal
toxic doses and trends of mortality caused will provide a
baseline system about the potential use of such botanical
compounds to be taken forward for insecticidal development.
Materials and methods
Insect collection and culturing
The study was conducted in insect ecology and behavior
laboratory of COMSATS University Islamabad (CUI), Paki-
stan during the year 2019–20. The laboratory cultures of
T. castaneum were started from beetles (ca. 500) collected
from the infested wheat storage facilities in Multan (Punjab)
Pakistan (+ 30° 11′ 52"N, + 71° 28′ 11" E). These adults
were cultured in wholemeal wheat flour with 5% brewer’s
yeast incubated at 25 ± 5ºC and 65 ± 5% r.h and let them
complete one generation in the laboratory. This was done to
remove any dietary variations in the field-collected beetles
as well as to get uniform age cultures for future studies. The
pupae developed were sex-sorted based on their abdominal
characteristics (Halstead 1963). For bioassays, F1 larvae of
the uniform age were used (3rd Instar).
Plant extracts preparation
After proper identification, healthy and disease-free seeds
of P. harmala and C. intybus were collected during Sep-
tember 2019, from Plant Genomic Research Institute
(PGRI) National Agriculture Research Council (NARC),
Islamabad-Pakistan. Identification was carried out by Dr.
Mustafa Sajid PGRI (NARC), Islamabad. Voucher speci-
mens (PH-AJ-001) for P. harmala and (CI-AJ-001) for C.
International Journal of Environmental Science and Technology

Fig. 2  Major chemical constitu-

ents of C. intybus, a Inulin, b
Alpha-amgrin, c Cichoric acid,
d Lactucin, e Caffeic acid and f
Cichoriin (Abbas et al. 2015)

intybus were deposited in the herbarium of the Department Toxicity bioassay

of Biosciences, COMSATS University Islamabad.
After collection seeds were washed with deionized water
thrice to remove any dirt and were shade dried at room tem-
perature for a minimum time period of two weeks. Seeds
were finally grinded with a sterilized mortar and pestle and a
steel grinder to make a fine powder. 10 gm seed powder was
dissolved in 100 mL of both ethanol and methanol of analyti-
cal grade in a ratio of (1:10) and kept at room temperature
for 3 days in a lid-containing container with gentle agitation.
Extracts were then filtered with a filtration assembly using
a filter paper with a pore size of 0.45 µm. The filtrate was
evaporated under reduced pressure, lyophilized, and stored
at 4 °C till further use.
For bioassays series of concentrations from pure extracts
were prepared using serial dilution method. Pure extracts
were reconstituted as the stock solution of concentration
100 mg/mL and were diluted, in respective solvents to pre-
pare the test concentrations i.e. 5, 10, 20, 40, 80 (× ­103 ppm)
to be used in bioassays.
Contact toxicity of each plant extract was tested on ­3rd instar
larvae using filter paper treatment methods. A volume of
500 µL of each test concentration was applied to filter paper
disc (9 mm ø) placed in plastic Petri dish and was air-dried
before transferring the larvae. The uniformly aged larvae of
­3rd instar were retrieved using a 500 µm mesh sieve. Ten of
those larvae were then released on each of the treated filter
paper discs and were provided with 1 g of wholemeal wheat
flour to minimize the mortality due to starvation. All the bio-
assays were replicated thrice and the data on larval mortality
was recorded 1, 2, 3, 5, 9, and 15 days after application.
Statistical analyses
The data from both bioassays were analyzed in terms of per-
cent mortality. To overcome the effect of natural mortality,
as observed in control treatments, the mortality of treated
individuals was corrected using Schneider – Orelli's formula
(Püntener 1981).

13

Corrected Mortality
Mortality in Treatment − Mortality in Control
=
100 − Mortality in Control
× 100
The data was transformed appropriately, where required,
to conform to the ANOVA assumptions. The data for larval
mortality was analyzed by using 2-way factorial ANOVA
with three replications using type of botanical and concen-
trations as fixed factors. Where the significant effect of dif-
ferent treatments and their interactions were observed the
means were subjected to posthoc pairwise comparisons
using Fisher’s Least Significant Difference Test with sig-
nificance level adjusted for multiple comparisons (Benjamini
and Hochberg 1995).
To determine the lethal dose that may kill 50% of the
pest population (­ LD50), dose–mortality (corrected) data were
subjected to the probit analysis using Origin Pro 9.1 SRO
statistical software (OriginLab Corp, USA).
Results and discussion
Larval toxicity of P. harmala and C. intybus
In terms of larval mortality, no significant differences
among ethanol and methanol extracts of P. harmala were
observed (F1, 24 = 0.04; p = 0.843), whereas, different con-
centrations of each extracts had a highly significant differ-
ence (F5, 24 = 72.90; p < 0.001). Both type of solvent used
to prepare different concentrations of P. harmala and the
Fig. 3  Mortality of T. cas-
taneum larvae treated with
ethanol (a, c) and methanol (b,
d) extracts of P. harmala (a,
b) and C. intybus (c, d). The
bars are means ± SEM and the
superscript letters atop each bar
represent the pairwise post hoc
comparisons between types of
solvent used (uppercase letters)
and between different concen-
trations for a specific solvent
type (lowercase letters). The
bars with different letters are
significantly different from each
other (p < 0.05; Tukey’s HSD)
13
International Journal of Environmental Science and Technology
concentrations were used to test against T. castaneum lar-
vae, we observed a significant interaction (F5, 24 = 3.40;
p = 0.018). The larval mortality increased significantly
with the increase in solvent concentration in comparison to
control. The highest mortality was observed in the larvae
treated with 40 × ­103 ppm and 80 × ­103 ppm of both etha-
nol and methanol extracts (Fig. 3a). Ethanol extracts at the
concentration of 20 × ­103 ppm caused significantly higher
mortality of the treated larvae as compared to same concen-
tration of methanol extracts; however, at the lowest dose of
5 × ­103 ppm, methanol extracts caused significantly higher
mortalities of the treated larvae (Fig. 3a).
The results for larval mortality due to C. intybus extracts
has clearly demonstrated that methanol extracts had higher
toxicity as compared to the ethanol extracts (F1, 24 = 7.54;
p = 0.011), in addition to a significant difference between
concentrations of each type of extract (F 5, 24 = 51.05;
p < 0.001). Moreover, a significant interaction among the
type of solvent used and the concentration of each type of
extract was observed (F5, 24 = 4.12; p = 0.008). The four
of the highest concentrations of ethanol extracts (i.e. 10
– 80 × ­103 ppm) had statistically similar toxicity in terms
of larval mortality and it was significantly higher than the
5 × ­103 ppm concentration (Fig. 3b).
The methanol extracts of C. intybus at different concen-
trations had significantly higher toxicities in comparison
to their corresponding concentrations of ethanol extracts,
except 10 × ­103 ppm where this trend was reversed (Fig. 3b).
The larvae treated with 20–80 × ­103 ppm concentrations of
methanol extracts had significantly highest mortality in com-
parison to 5 and 10 × ­103 ppm concentrations (Fig. 3b).
International Journal of Environmental Science and Technology

Similarly, the ethanol extracts of C. intybus had an
increasing trend in larval toxicity and on the 9th day after
application, a significant difference in larval survival was
observed (F5, 17 = 4.25; p < 0.019) (Fig. 4c). More than
50% larvae died on the 15th day after dose application,
except those treated with 5 × ­103 ppm dosage (Fig. 4c).
For methanol extracts of C. intybus, all concentrations had
significantly different survival of treated larvae in compari-
son to control from 3­ rd day after application (F5, 17 = 6.17;
p = 0.005). No significant difference in larval survival was
observed between larvae treated with different populations
up to the 9th day after dose application (Fig. 4d). On 15th
day post dose application, significantly lowest survivals
(F5, 17 = 67.43; p < 0.001) were observed in the population
treated with 20, 40, and 80 × ­103 ppm concentrations of etha-
nol extracts, followed by the other two lowest concentrations
(Fig. 4d).
The efficacy of ethanol and methanol extracts of both P.
harmala and C. intybus in terms of acute larval toxicity is
provided in Tables 1 and 2, respectively. The results have
clearly demonstrated that toxicity of both botanical extracts
increased as the exposure time increased. The ­LD50 values
for ethanol extracts of P. harmala were significantly lower
in comparison to those for methanol extracts (Table 1). The
­LD50 values decreased significantly as the time after expo-
sure increased. Just one day after application the ­LD50 value
for ethanol extract was determined to be 300.6 × ­103 ppm,
which is significantly higher than the ­LD50 value on the 15th
day after application (7.7 × ­103 ppm) (Table 1). The results
from acute larval toxicity caused because methanol extracts
of P. harmala demonstrated a strong correlation with expo-
sure time (Table 1). A significantly higher L ­ D50 value was
observed just one day after application (i.e. 1219 × ­103 ppm)
but on the 15th day after treatment, it declined sharply to a

Fig. 4  Larval survival of T.

castaneum larvae treated with
ethanol (a, c) and methanol (b,
d) extracts of P. harmala (a, b)
and C. intybus (c, d). The bars
are means ± SEM. The data
points where the SEM overlaps
are not statistically different
from one another (p < 0.05;
Tukey’s HSD)

Table 1  The ­LD50 values of ethanol and methanol extracts of P. harmala on T. castaneum larvae at different exposure times
Days after n Ethanol extracts Methanol extracts
application 3
LD50 (× ­10 ppm) Confidence interval Correlation LD50 (× ­103 ppm) Confidence interval Correlation
(× ­103 ppm) coefficient (× ­103 ppm) coefficient

1 180 300.6 215.3–418.8 0.4596 1219.0 679.2–2187.8 0.3980

2 180 146.9 116.7–185.4 0.6862 527.2 354.0–785.2 0.5175
3 180 73.8 56.2–97.1 0.5725 134.3 103.8–173.8 0.7673
5 180 65.5 49.7–86.1 0.4210 110.9 83.2–148.3 0.5322
9 180 43.3 30.5–61.4 −0.0275 54.6 37.3–79.8 −0.2183
15 180 7.7 5.3–11.1 0.4437 8.8 6.9–11.2 0.8499

13
 International Journal of Environmental Science and Technology

Table 2  The ­LD50 values of ethanol and methanol extracts of C. intybus on T. castaneum larvae at different exposure times
Days after n Ethanol extracts Methanol extracts
treatment 3
LD50 (× ­10 ppm) Confidence interval Correlation LD50 (× ­103 ppm) Confidence interval Correlation
(× ­103 ppm) coefficient (× ­103 ppm) coefficient

1 180 341.2 182.4–636.8 –0.6717 386.4 252.9–588.8 0.1326

2 180 158.5 113.5–221.3 0.4054 273.5 181.1–413.0 0.2264
3 180 92.7 61.7–139.3 –0.5347 133.4 88.5–201.4 0.4694
5 180 99.1 66.4–148.3 –0.4128 200.4 125.6–319.9 –1.6290
9 180 37.6 21.7–65.3 –2.5090 56.6 36.6–87.7 –0.3217
15 180 7.9 6.1–10.3 0.7358 12.6 8.1–19.7 –0.1024

surprising concentration i.e. 8.8 × ­103 ppm (Table 1), which
is significantly similar to the corresponding ­LD50 value of
ethanol extracts.
A very identical trend in decrease in L ­ D50 value for
ethanol and methanol extracts of C. intybus was observed
(Table 2). Although no significant difference among cor-
responding ­LD50 values for ethanol and methanol extracts
was observed, a significant decrease in ­LD50 values has been
clearly demonstrated along the exposure time (Table 2). One
day after application the L­ D50 value for ethanol and metha-
nol extracts were 341.2 × ­103 ppm and 386.4 × ­103 ppm
respectively (Table 2). The toxicity significantly increased
until the 15th day after application and the lowest ­LD50 val-
ues were recorded as 7.9 × ­103 ppm and 12.6 × ­103 ppm for
ethanol and methanol extracts, respectively (Table 2).
Discussion
P. harmala, is a bushy perennial shrub. Earlier studies
have stated the efficacy of P. harmala against T. castaneum
(Jbilou et al. 2006; Jbilou and Sayah 2007) and aphid spe-
cies (Salari et al. 2012), and our findings complement the
previous findings as well.
C. intybus is a medicinally important plant and has been
used for centuries for primary health care needs. Based on
its indigenous and exogenous introductions it has a long his-
tory of therapeutic use (Gurib-Fakim 2006; Li et al. 2011).
Our studies have demonstrated that natural extracts of the
plants have a tendency to control T. castaneum in stored
wheat grains.
The efficacy of C. intybus plant extracts against T. cas-
taneum larvae (Pascual-Villalobos and Robledo 1998) and
dipterous larvae (Mansour et al. 2011, 2014) has been
reported. Our study is unique in the sense that, seeds of
C. intybus have not been tested so far for their insecticidal
activity, which is carried out in current study.
Both P. harmala and C. intybus plants have demon-
strated efficacy against T. castaneum larvae, although
no knockdown effect was observed (Fig. 4). Both plants
had shown better efficacy when their constituents were
extracted in ethanol as compared to methanol with an
incremental effect with the increase in concentration i.e.
from 5 × ­1 0 3 ppm to 80 × ­1 0 3 ppm. For C. intybus, our
results are different from the previously reported studies
of the root extracts of C. intybus (Ali et al. 2018), the
methanol extract exhibited strong insecticidal activity in
comparison to ethanol extract, but in our investigations,
the reverse is true and the reason may be the fact that we
have dealt with seeds instead of roots of C. intybus, the
ethanol extract showed strong larvicidal activity in com-
parison to methanol extract.
In detailed investigations, it was concluded that metha-
nol and ethanol extracts of P. harmala (seeds) and C. inty-
bus (seeds) had good antimicrobial and antioxidant activ-
ity and can be used for further drug discovery in the quest
of novel antimicrobial and antioxidant drugs. The plausi-
ble compound/s responsible for the bioactivity were also
identified in the said dissertation (Javaid A, 2021). Using
various chromatographic and spectroscopic techniques
the most abundant compounds in the seeds of P. harmala
and C. intybus were identified using GC–MS and the com-
pounds responsible for strong antioxidant potential in seeds
of these plants were also ruled out using Orbitrap LC–MS.
It was found that P. harmala seeds were rich in flavonoids
like harmine and harmaline, fatty acid derivatives, esters,
etc. and these compounds are responsible for a number of
pharmacological activities of these plants i.e., antioxidant,
insecticidal activity, etc. The Flavonoids have a catecholic
B ring that is responsible for the insecticidal activity of P.
harmala (Onyilagha et al. 2004). As flavonoids hold great
structural diversity thus a range of activity is expected from
them depending upon their structural diversity.
Upon GC–MS of C. intybus, the most abundant com-
pounds in seed extracts were found to be fatty acids like

13
International Journal of Environmental Science and Technology
9-cis-12-cis-octadecadienoic acid and esters i.e. (E, E,
E)-9-octadecenoic acid-1,2,3 propanetriyl ester. So, the
presence of these bioactive compounds has been confirmed
in the dissertation (Javaid A, 2021). These compounds are
possibly responsible for natural insecticidal activity of C.
intybus against T. castaneum larvae.
Although, T. castaneum is known to have excellent
olfactory capabilities (Padin et al. 2013) and their genome
sequences reveal a large number of odorant receptor genes
compared to those found in other insects whose olfactory
genomes have been studied (Ahmad et al. 2013). These
genes enable these animals to detect and avoid toxic com-
pounds. But this phenomenon had not been observed dur-
ing bioassays where the mortality of larvae increased with
the increase in concentrations. These findings significantly
propose the inability of the T. castaneum larvae to avoid
toxic metabolites using their olfactory senses. Hence, posi-
tively suggest the use of botanical extracts of P. harmala
and C. intybus for stored grain protection.
Conclusion
Being eco-friendly and bio-degradable product, P. har-
mala and C. intybus extracts may provide a complemen-
tary management method to limit chemical control in
grain storage facilities and food processing industries.
The use of these botanicals can augment synthetic insec-
ticide efficiencies. Moreover, their use is safe, environ-
ment friendly, cheap, and versatile. Further investigation
in dose–response relationship can be helpful in developing
easy and effective application technologies for pest man-
agement in domestic and small-scale field storage, as well
as large-scale storage houses.
Acknowledgements The authors are thankful to Higher Education
Commission of Pakistan and their parent organizations for providing
infrastructure and support to conduct the study.
Author’s contributions ZA and FA Conceptualized and designed
experiments in the project. AJ, TZ, FA, SN, MM and QS performed
experimental work, data Curation and analysis. FA & SN did Writing,
review and editing of the manuscript. ZA finally reviewed and approved
the manuscript.
Funding No funding was received for conducting this study.
Availability of data and materials The datasets used and/or analysed
during the current study are available from the corresponding author
on reasonable request.
Code availability Not applicable.
Declarations
Conflict of interest The authors have no conflicts of interest to declare
that are relevant to the content of this article.
Ethical approval Not applicable.
Consent to participate Not applicable.
Consent for publication Not applicable.
References
Abbas ZK, Saggu S, Sakeran MI, Zidan N, Rehman H, Ansari AA
(2015) Phytochemical, antioxidant and mineral composition of
hydroalcoholic extract of chicory (C. intybus L.) leaves. Saudi J
Biol Sci 22:322–326
Abbassi K, Atay-Kadiri Z, Ghaout S (2003) Biological effects of alka-
loids extracted from three plants of Moroccan arid areas on the
desert locust. Physiol Entomol 28:232–236
Ahmad I et al (2013) Spatio-temporal variations in some medicinally
important biochemical constituents of P. harmala (Hermal). Pak
J Bot 45:601–607
Al-Allaf TA, Rashan LJ (1998) Synthesis and cytotoxic evaluation of
the first trans-palladium (II) complex with naturally occurring
alkaloid harmine. Eur J Med Chem 33:817–820
Al-Shamma A et al (1981) Antimicrobial agents from higher plants.
Antimicrobial agents from P. harmala seeds. J Nat Prod
44:745–747
Ali SI, Gopalakrishnan B, Venkatesalu V (2018) Chicory (Cichorium
intybus) and wormwood (Artemisia absinthium) extracts exhibit
strong larvicidal activity against mosquito vectors of malaria, den-
gue fever, and filariasis. Parasitol Int 67:781–786
Ameziane N, Boubaker H, Boudyach H, Msanda F, Jilal A, Benaoumar
AA (2007) Antifungal activity of Moroccan plants against citrus
fruit pathogens. Agron Sustain Dev 27:273–277
Arnason J, Philogène B, Morand P (1990) Insecticides of plant origin.
Chem Inform 21
Arroyo M (1995) Lucha contra las plagas y proteccion de los cul-
tivo: una approximacion historica. Conferencias del Seminario
de Fitopatologia, pp 41–51
Arshad N, Neubauer C, Hasnain S, Hess M (2008) P. harmala can mini-
mize Escherichia coli infection in poultry, but long-term feeding
may induce side effects. Poult Sci 87:240–249
Asghari G, Lockwood GB (2002) Stereospecific biotransformation of
(±) phenylethyl propionate by cell cultures of P. harmala L. Iran
Biomed J 6:43–46
Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate:
a practical and powerful approach to multiple testing. J R Stat Soc
Series B Stat Methodol 57:289–300
Bughio F, Wilkins R (2004) Influence of malathion resistance status on
survival and growth of T. castaneum (Coleoptera: Tenebrionidae),
when fed on flour from insect-resistant and susceptible grain rice
cultivars. J Stored Prod Resn 40:65–75
Bukhari N, Choi J, Jeon C, Park H, Kim W, Khan MA, Leet S (2008)
Phytochemical studies of the alkaloids from P. harmala. Appl
Chem 12:101–104
Carlini CR, Grossi-de-Sá MF (2002) Plant toxic proteins with insecti-
cidal properties. A review on their potentialities as bioinsecticides.
Toxicon 40:1515–1539
Cox P (2004) Potential for using semiochemicals to protect stored prod-
ucts from insect infestation. J Stored Prod Res 40:1–25
13

Ehsanpour A, Saadat E (2002) Plant regeneration from hypocotyl cul-
ture of P. harmala. Pak J Bot 34:253–256
El Bahri L, Chemli R (1991) P. harmala L: a poisonous plant of North
Africa. Vet Hum Toxicol 33:276–277
Fan B et al (1997) Effect of total alkaloid of P. harmala L. in the treat-
ment of experimental haemosporidian infections in cattle. Trop
Anim Health Prod 29:77S-83S
Gurib-Fakim A (2006) Medicinal plants: traditions of yesterday and
drugs of tomorrow. Mol Aspects Med 27:1–93
Halstead DGH (1963) External sex differences in stored-products
Coleoptera. Bull Entomol Res 54:119–134
Harsh ML, Nag TN (1984) Antimicrobial principles from in vitro tissue
culture of P. harmala. J Nat Prod 47:3 65–367
Isman MB (2006) Botanical insecticides, deterrents, and repellents in
modern agriculture and an increasingly regulated world. Annu
Rev Entomol 51:45–66
Jacobson M (1982) Plants, insects, and man—their interrelationships.
Econ Bot 36:346–354
Javaid A (2021). Functional profiling of bioactive compounds from
indigenous plant resources. PhD Dissertation, COMSATS Uni-
versity Islamabad.
Javaid A, Anwar SA, Z, Naseem, S, (2021) Chromatographic and Spec-
troscopic Fingerprinting of Ficus carica and Evaluation of In Vitro
Antioxidant Activity. Int J Agri Biol 25:677–682
Jbilou R, Ennabili A, Sayah F (2006) Insecticidal activity of four
medicinal plant extracts against T. castaneum (Herbst)(Coleop-
tera: Tenebrionidae). African J Biotech 5:936–945
Jbilou R, Sayah F (2007) Effects of P. harmala (Zygophyllaceae) seed
extracts on the development of T. castaneum (Coleoptera: Ten-
ebrionidae). Int J Trop Insect Sci 27:199–209
Jilani G, Amir P (1987) Economics of neem [Azadirachta indica] in
reducing wheat storage losses: policy implications [Pakistan].
SEARCA [Southeast Asian Regional Center for Graduate Study
and Research in Agriculture] Tech Bull (Philippines)
Jilani G, Saxena R, Rueda B (1988) Repellent and growth-inhibiting effects
of turmeric oil, sweetflag oil, neem oil, and “Margosan-O” on red flour
beetle (Coleoptera: Tenebrionidae). J Econ Entomol 81:1226–1230
Kartal M, Altun M, Kurucu S (2003) HPLC method for the analysis
of harmol, harmalol, harmine and harmaline in the seeds of P.
harmala L. J Pharm Biomed Anal 31:263–269
Khaliq A, Sagheer M, Javed M (2014) Estimation of quality deteriora-
tion in different rice genotypes infested by T. castaneum (Herbst)
under a biotic stress. Cercet Agro Mold 47:47–56
Kheradpir N (2014) Food preference of T. castaneum among four flour
types. Euro J Exp Biol 4:436–439
Kirtikar KR, Basu BD (2012) Indian Medicinal Plant, 2nd edn. Inter-
national book distributer, India
Kubo I (2006) New concept to search for alternate insect control agents
from plants. Adv Phytomed 3:61–80
Lamchouri F et al (1999) Antitumour principles from P. harmala seeds.
Therapie 54:753–758
Li Y et al (2011) Tumor-derived autophagosome vaccine: mechanism
of cross-presentation and therapeutic efficacy. Clin Cancer Res
17:7047–7057
Mansour S, Bakr R, Mohamed R, Hasaneen N (2011) Larvicidal activity
of some botanical extracts, commercial insecticides and their binary
mixtures against the housefly, Musca domestica L. Seeds 4:1–13
Mansour SA, Ibrahim RM, El-Gengaih i SE, (2014) Insecticidal
activity of chicory (C. intybus L.) extracts against two dipterous
insect-disease vectors: mosquito and housefly. Ind Crops Prod
54:192–202
Nandagopal S, Kumari BR (2007) Phytochemical and antibacterial
studies of Chicory (C. intybus L.)-A multipurpose medicinal
plant. Adv Biol Res 1:17–21
13
International Journal of Environmental Science and Technology
Nawrot J, Harmatha J (2012) Phytochemical feeding deterrents for stored
productinsect pests. Phytochem Rev 11:543–566
Niroumand MC, Farzaei MH, Amin G (2015) Medicinal properties of P.
harmala L. in traditional Iranian medicine and modern phytotherapy:
a review. J Tradit Chin Med 35:104–109
Onyilagha JC, Lazorko J, Gruber MY et al (2004) Effect of flavonoids on
feeding preference and development of the crucifer pest Mamestra
configurata Walker. J Chem Ecol 30:109–124
Padin SB, Fuse C, Urrutia MI, Dal Bello GM (2013) Toxicity and repel-
lency of nine medicinal plants against T. castaneum in stored wheat.
Bull Insectology 66:45–49
Pascual-Villalobos MJ, Robledo A (1998) Screening for anti-insect activity
in Mediterranean plants. Industrial Crops and Products 8:183–194
dPitre S, Srivastava SK (1987) Two new anthraquinones from the
seeds of P. harmala. Planta Med 53:106–107
Püntener W (1981) Manual for field trials in plant protection. Ciba-Geigy,
Pushparaj P, Low H, Manikandan J, Tan B, Tan C (2007) Anti-diabetic
effects of C. intybus in streptozotocin-induced diabetic rats. J Ethnop-
harmacol 111:430–434
Rates SMK (2001) Plants as source of drugs. Toxicon 39:603–613
Salari E, Ahmadi K, Dehyaghobi RZ, Purhematy A, Takalloozadeh HM
(2012) Toxic and repellent effect of harmal (P. harmala L.) ascetonic
extract on several aphids and T. castaneum (Herbst). Chil J Agric Res
72:147–151
Sathiyamoorthy P, Lugasi-Evgi H, Van-Damme P, Abu-Rabia A, Gopas
J, Golan-Goldhirsh A (1997) Larvicidal activity in desert plants of
the Negev and Bedouin market plant products. Int J Pharmacogn
35:265–273
Secoy D, Smith A (1983) Use of plants in control of agricultural and
domestic pests. Econ Bot 37:28–57
Shad M, Nawaz H, Rehman T, Ikram N (2013) Determination of some bio-
chemicals, phytochemicals and antioxidant properties of different parts
of C. intybus L.: A comparative study. J Anim Plant Sci 23:1060–1066
Shahverdi AR, Monsef-Esfahani HR, Nickavar B, Bitarafan L,
Khodaee S, Khoshakhlagh N (2005) Antimicrobial activity and
main chemical composition of two smoke condensates from P.
harmala seeds. Zeitschrift Für Naturforschung C 60:707–710
Sharaf M, El-Ansari MA, Matlin SA, Saleh NA (1997) Four flavonoid
glycosides from P. harmala. Phytochem 44:533–536
Siddiqui S, Khan OY, Siddiqui BS, Faizi S (1987) Harmalidine, A
β-carboline alkaloid from P. harmala. Phytochem 26:1548–1550
Sobhani AM, Ebrahimi S-A, Mahmoudian M (2002) An in vitro evalu-
ation of human DNA topoisomerase I inhibition by P. harmala L.
seeds extract and its beta-carboline alkaloids. J Pharm Pharma-
ceutical Sci 5:19–23
Tahraoui A, El-Hilaly J, Israili Z, Lyoussi B (2007) Ethnopharma-
cological survey of plants used in the traditional treatment of
hypertension and diabetes in south-eastern Morocco (Errachidia
province). J Ethnopharmacol 110:105–117
UNEP (2003) Declaration on methyl bromide. Report of the Fifteenth
Meeting of the Parties to the Montreal Protocol on substances
that deplete the ozone layer Annex VIII, Nairobi. 10–14 No 2003.
http://​ozone.​unep.​org/​Publi​catio​ns/​MP_​Handb​ook/​Secti​on_3.8​ _​
Annexe​ s_D​ eclar​ ation​ s/D
​ eclar​ ation_o​ n_M
​ eBr.s​ html, Last updated
on September 17, 1997. Accessed on July 24, 2012
Yousefi R, Ghaffarifar F, Asl AD (2009) The effect of Alkanna tinc-
turia and P. harmala extracts on Leishmania major (MRHO/
IR/75/ER) in vitro. Iran J Parasitol 4:40–47
Zaker F, Oody A, Arjmand A (2007) A study on the antitumoral and
differentiation effects of P. harmala derivatives in combination
with ATRA on leukaemic cells. Arch Pharmacal Res 30:844–849