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Global Research journal of Natural Science  
& Technology (GRJNST)  
Volume: 04 - Issue 2 (2026), 2066  
ISSN P: 2790-7643 ISSN E: 2790-7651  
www.grjnst.net  
https://doi.org/10.53762/grjnst.04.02.17  
Identification of volatile constituents and biological activities of Sophora  
alopecuroides of Balochistan  
Received: 24 December 2025. Accepted: 26 February 2026. Published: 20 April 2026  
Asghar Ali  
Department of Chemistry  
University of Balochistan Quetta, Pakistan  
Samar Ali  
Department of Chemistry  
University of Balochistan Quetta, Pakistan  
Nimra Fazal  
Department of Chemistry  
University of Balochistan Quetta, Pakistan  
Sara Zameer  
Department of Chemistry  
University of Balochistan Quetta, Pakistan  
Shazia Saeed  
Department of Botany  
University of Balochistan Quetta, Pakistan  
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Abstract: Sophora alopecuroides belongs to genus Sophora and family Fabaceae. It consists of  
approximately 60 to 70 species. The present research work was performed for the identification of  
volatile constituents along with antibacterial and anti-inflammatory activities of Sophora alopecuroides.  
The plant was collected from Kalat, Balochistan. The n-hexane fraction of the plant was evaluated for  
the determination of volatile compounds by using gas chromatography-mass spectrometry (GC-MS).  
The fourier-transform infrared spectroscopy (FTIR) was used to identify the functional groups of the  
compounds. The methanolic extract of the plant was evaluated for anti-inflammatory and antibacterial  
by using chemiluminescence protocol and agar well diffusion method respectively. The GC-MS analysis  
showed the presence of 34 volatile compounds. The FTIR spectra of methanolic extract showed the  
presence of functional groups such as alcohol, alkane, carbonyl compound and ester. The plant extract  
showed moderate antibacterial activity against gram positive Staphylococcus aureus and gram negative  
Escherichia coli bacterial strains, and with zones of inhibition of 19 mm and 21 mm respectively. The  
plant extract exhibited strong activity IC50 56.14± 2.82 for anti-inflammatory activity against standard  
drug ibuprofen. The plant showed the presence of significant number of volatile compounds which  
belongs to different classes of compounds. It is further suggested that Sophora alopecuroides may  
further be studied for the isolation of naturally occurring compounds and more pharmacological  
activities.  
Key words: GC-MS, Antibacterial activity, Anti-inflammatory activity, Sophora alopecuroides  
Introduction  
Sophora alopecuroides a member of genus Sophora and belongs to the family Fabaceae. It is medicinally important  
plant and grows in Kalat and Quetta, Balochistan. It is widely distributed in Pakistan, China, Iran, Afghanistan,  
Turkey and Kazakhstan (Zhao et al., 2023).  
Sophora alopecuroides is a medicinally important plant. It is used to treat various disease like dysentery,  
eczema,furuncle, recurrent dermatitis, infectious disease and cancer by local population (Pervez et al., 2018). The  
previous study reported the presence of quinolizidine alkaloids, in particular, matrine, oxymatrine, sophocarpine  
and sophoridine. These alkaloids have shown strong anti-inflammatory, antiviral, antibacterial, antifungal, anti-  
tumor, and insecticidal properties making the plant significant in both the traditional and modern medicine.  
Matrine and oxymatrine have significant anticancer properties that cause tumor cell death (apoptosis), tumor  
angiogenesis, and cancer cell growth (He et al., 2016).  
This present research work was focused on the identification of volatile compounds of Sophora alopecuroides.  
The Balochistan province is very rich in medicinal plants and the local population uses the various plants to treat  
different diseases. This study will help to identify the volatile compounds and assess the biological activities and  
it can lead to potential drug discovery.  
Plant collection: The medicinal plant Sophora alopecuroides was collected from Kalat, Balochistan.  
Plant Identification: Sophora alopecuroides was identified with the help of Flora of Pakistan and confirmed by  
Dr. Shazia Saeed, Department of Botany, University of Balochistan Quetta. The specimen was deposited at the  
University of Balochistan Herbarium with a voucher number UOB-000245.  
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Methodology  
Extraction process of Medicinal plant Sophora alopecuroides  
The plant (5 Kg) was thoroughly washed and shade dried. After drying, it was ground into a powder. The material  
was soaked in 10 L of methanol for 7 days and this process was repeated three times at room temperature. The  
collected methanolic extract was filtered using Whatman filter paper and concentrated using a rotary evaporator  
at 45 oC. The crude methanolic extract was then freeze dried at 4 oC to get gummy solid extract.  
Fractionation of Crude Methanolic Extract  
The crude methanolic extract was suspended in distilled water to get an aqueous solution. It was then fractionated  
into n-hexane fraction using a separating funnel.  
GCMS Analysis  
The n-hexane fraction of the Sophora alopecuroides was subjected to GCMS analysis. The GC column using 5%  
phenyl methyl siloxon as the stationary phase was used for this purpose. The sample gaseous phase with pressure  
and velocity carried by the inert helium gas. The equipment fitted with a HP-5MS capillary column. Helium gas  
was taken as the carrier gas at a flow of 1 mL/min. The temperature of the oven was programmed between 60oC  
to 280oC. The mass spectrum was run in EI mode at 70 eV with a scan range of m/z 50-500 and NIST library  
was used to identify compounds (Mondello et al., 2008)  
FTIR Analysis  
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FTIR was used for the detection of classes of compounds on the basis of functional groups. The resolution used  
-1  
-1  
4 cm  
in the 4000-400 cm  
range and32 scans with a DTGS detector at room temperature. The  
characteristic absorption peaks were used to identify functional groups. (Stuart et al., 2004)  
Biological Activities  
Antibacterial activity  
The preparation of a bacterial culture was done by inoculation of a particular bacterial strain in sterile nutrient  
broth in a 1.5 ml flask. The culture was kept at 37oC overnight for the growth of bacteria. The nutrient was added  
into petri dishes, which were left to solidify after incubation. A 100 µL agar suspension was spread all over the  
agar using a sterile swab with the bacterial suspension. A sterile pipette tip was used to pierce the agar with wells  
of about 68 mm diameter. The test samples, positive and negative controls, were put in each well. The incubation  
was done at 37 oC and given 18-24 hours. The zones of inhibition surrounding each well were then measured in  
millimeters by means of a ruler or caliper after incubation (Vlagas et al., 2007).  
Anti-inflammatory activity  
The anti-inflammatory activity was measured using chemiluminescense assay of a sample based on its capacity  
to inhibit the release of reactive oxygen species (ROS) by stimulated neutrophils.The test samples were  
initially dissolved in dimethyl sulfoxide (DMSO) or any other appropriate solvent to make the necessary  
concentrations (Allen et al., 1986).Then, neutrophils, luminol, and the test sample were mixed together in a  
tube or microplate well of a luminescence meter.Then, an activator was incorporated to activate the  
neutrophils. A control sample was also prepared, that was, without the test compound. The resulting light  
(chemiluminescense) was promptly measured with a luminescense meter within 15-30 minutes at 37oC.When  
the light emission was decreased by the test sample, then it implied that the latter was preventing the formation  
of reactive oxygen species, which signifies anti-inflammatory action.The inhibition percent was determined  
by dividing the luminescence of the treated sample by the control (Dahlgren et al., 1999).  
Result and Discussion  
GC-MS analysis  
The n-hexane fraction of Sophora alopecuroides was analyzed through GC-MS method. The findings of the  
current work revealed the existence of 34 compounds including 5-Hepten-2-one, 7-phenyl-, O-methyl oxime,  
oxime derivative act as enzyme-interaction potential. 2-Methoxy-4-vinylphenol are phenolic compound act as  
antimicrobial (Islam etal.,2018). 6,8-Dioxa-3-thiabicyclo(3,2,1)octane 3,3-dioxide and Thiophene, tetrahydro-2-  
methyl- are sulfur & oxygen heterocyclic compound act as anti-inflammatory. Hentriacontane-10,14,16-trione and  
Cyclopentane, 1,2-dimethyl-3-(1-methylethyl) are cyclic hydrocarbon and long chain alkane Hexacosane,  
Octacosane, Eicosane, Tetratetracontane. (Shabana et al., 2012). 2-Hexadecene-3,7,11,15-tetramethyl and  
Heptadecane, 2,6,10,15-tetramethyl, Bicyclo[3.1.1]heptane, 2,6,6-trimethyl-, Phytol, Phytol acetate, Squalene are  
terpenoids and act as insecticide (Islam etal.,2018). 9-Octadecyne, and 1,4-Eicosadiene are Unsaturated  
Hydrocarbon act as lipid extract. (Ling etal.,1998). 2-O-Methyl-D-mannopyranose and 4-O-Methyl-D-  
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arabinose, Methyl-β-D-arabinopyranoside, 1,2-Cyclohexanedicarboxylic acid are sugars and act as metabolic  
intermediate (Dewicketal.,2009). 9,12-Octadecadienoic acid (Linoleic acid), 7,10,13-Hexadecatrienoic acid,  
9,11-Octadecadiynoic acid, 8-hydroxy methyl ester, Methyl stearate and Butanoic acid, 3-methyl-, 3,7-dimethyl-  
6-octenyl ester are poly saturated fatty acid act as antioxidant and antimicrobial (Ling et al., 1998). Sophoramine  
and Thioctic acid are alkaloid and organic acid, act as cytotoxic and neuroactive. Stigmastan-3,5-diene, Α-  
Ergostenol, and Stigmasterol, Cholest-2-ene-2-methanol are steroid derivatives act as hypocholesterolemic (Packer  
etal.,1995). Vitamin E are (α-tocopherol) act as fat-soluble vitamin. (Brigelius et al., 1999)  
The results are shown in table 1 and chromatogram and mass spectrum of each volatile compound shown in figure  
1 & 2.  
Table.1. List of volatile compounds identified using GC-MS analysis  
S. No  
Name of compound  
Retention  
time  
%Area  
sum  
Molecular  
formula  
Molecular  
weight  
1
2
3
5-Hepten-2-one, 7-phenyl-, O-methyl oxime  
2-Methoxy-4-vinylphenol  
2.238  
3.471  
6.474  
0.43  
0.21  
8.09  
C14H19NO  
217  
150  
164  
C
9H2O10  
6,8-Dioxa-3-thiabicyclo(3,2,1)octane 3,3-  
dioxide  
C
5H  
8
O4  
S
4
5
Hentriacontane-10,14,16-trione  
2-Hexadecene, 3,7,11,15-tetramethyl  
Bicyclo[3.1.1]heptane, 2,6,6-trimethyl-  
Cyclopentane, 1,2-dimethyl-3-(1-methylethyl  
9-Octadecyne  
6.903  
8.335  
8.419  
8.679  
8.867  
9.410  
9.688  
0.23  
0.15  
1.76  
0.18  
1.37  
7.63  
C31H58O3  
C20H38  
C10H18  
C10H20  
C18H34  
478  
278  
138  
140  
250  
194  
304  
6
7
8
9
2-O-Methyl-D-mannopyranosa  
C
7H14O6  
10  
9,11-Octadecadiynoic acid, 8-hydroxy methyl  
ester  
5.04  
C19H28O3  
11  
12  
13  
14  
15  
16  
17  
18  
19  
20  
21  
22  
Thiophene, tetrahydro-2-methyl-  
4-O-Methyl-d-arabinose  
Methyl-.beta.-D-arabinopyranoside  
9,12-Octadecadienoic acid  
7,10,13-Hexadecatrienoic acid  
Phytol  
9.827  
10.776  
10.818  
11.072  
11.150  
11.295  
11.495  
19.573  
20.546  
21.513  
22.322  
23.585  
3.14  
5.08  
8.75  
0.38  
1.00  
0.61  
0.23  
0.27  
0.29  
0.24  
0.19  
0.39  
C
5H10S  
102  
164  
164  
280  
250  
296  
298  
244  
296  
410  
619  
396  
C
6H12O5  
6H12O5  
C
C18H32O2  
C16H26O2  
C20H40  
Methyl stearate  
C19H38O2  
C15H20N2  
C21H44  
Sophoramine  
Heptadecane, 2,6,10,15-tetramethyl  
Squalene  
C30H50  
Tetratetracontane  
C44H90  
Stigmastan-3,5-diene  
C29H48  
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23  
24  
25  
26  
27  
28  
29  
30  
31  
32  
33  
34  
Hexacosane  
23.845  
24.038  
24.219  
24.666  
24.836  
25.198  
25.754  
26.449  
26.739  
27.730  
27.796  
29.349  
0.36  
2.51  
0.37  
0.14  
0.32  
0.15  
0.16  
0.74  
0.98  
1.52  
0.45  
0.28  
C26H54  
C29H50O2  
C22H44O2  
C28H48O  
C29H48O  
C28H58  
366  
430  
338  
400  
412  
394  
414  
258  
278  
206  
172  
238  
Vitamin E  
Phytol, acetate  
Alpha.-Ergostenol  
Stigmasterol  
Octacosane  
Cholest-2-ene-2-methanol  
Eicosane  
C28H46O  
C20H42  
1,4-Eicosadiene  
Thioctic acid  
C20H38  
C
8H14O2  
8H12O4  
C15H26O2  
S
2
1,2-Cyclohexanedicarboxylic acid,  
C
Butanoic acid, 3-methyl-, 3,7-dimethyl-6-  
octenyl ester  
These volatile constituents are classified on the basis of chemical structure, functional group including hydrocarbon  
4.12%, fatty acid derivatives 8.45%, carbohydrates 21.46%, steroids 1.01%, terpenoids 3.13%, alkaloids 0.27%,  
oxidative property 2.51%, cycloalkanes 0.63%, phenolic compound 0.21%, sulphur containing compound  
11.23%, ketone 0.66 %.  
Figure.1. GC-MS Chromatogram of n-hexane fraction of Sophora alopecuroides  
Figure.2. Mass Spectrum of each volatile compounds identified inSophoraalopecuroides  
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FTIR analysis  
FTIR (Fourier Transform Infrared) was used to identify the functional groups in the methanolic extract of the  
plant. The spectrum confirmed the presence of alcohol, alkane, carbonyl compound and ester. The strong peak at  
3267.78cm⁻¹ confirmed an O-H stretching vibration, which is typical peak of an alcohol functional group. The  
peak at the 1634.45 cm⁻¹, which denote stretching of carbonyl group C=O. The presence of alkane was verified  
by the peak appeared at 1406.47 cm⁻¹ confirmed C-H bending vibrations. The peak appeared at 1009.41cm⁻¹,  
which was stretching of ester. The results are shown in table 2 and IR spectrum of methanolic extract in figure 3.  
Table.2.IR spectrum of Sophora alopecuroides obtained through FTIR analysis  
Functional group  
Alcohol / phenol  
carbonyl group  
Alkane  
General formula  
O–H  
Vibrational modes  
Stretching  
Methanolic extract  
3267.78  
C=O  
C–H  
Stretching  
Bending  
1634.45  
1406.47  
Ester  
RCOOR  
Stretching  
1009.41  
Figure. 3. IR spectrum of Methanolic extract of Sophora alopecuroides  
Antibacterial activity  
The antibacterial activity of the methanolic extract of Sophora alopecuroides was tested by using agar well  
diffusion method against gram positive Staphylococcus aureus and gram negative Escherichia coli bacterial  
strains. The plant extract showed moderate antibacterial effect on Staphylococcus aureus and Escherichia coli  
with zones of inhibition of 19mm and 21mm respectively. The concentration of standard drug Ofloxacin  
was 50 μg/ml and it showed 26mm zone of inhibition. The results are shown in Table 3 & Figure 4.  
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Table. 3. Antibacterial activity in medicinal plant Sophora alopecuroides was determined in methanolic extract.  
Name of Bacterial stains  
Inhibition of  
extract  
Inhibition of drug  
Staphylococcus aureus (NCTC 13277)  
Escherichia coli (ATCC 25922)  
(19mm)  
(21mm)  
(26mm)  
(26mm)  
Figure. 4. Comparison of antibacterial activity of inhibition of compound and inhibition of drug  
Anti-inflammatory activity  
The methanolic extract of Sophora alopecuroides was evaluated for the anti-inflammatory activity using  
Chemiluminescence protocol. The extract was tested against standard drug Ibuprofen. The methanolic extract  
concentration exhibited 78.04% zone of inhibition and IC50± SD μg/ml was 56.14± 2.82. The standard drug  
Ibuprofen concentration was 25μg/ml and showed 73.2 zone of inhibition and the standard drug IC50± SD  
μg/ml was 11.2±1.9 μg/ml. The methanolic extract of medicinal plant exhibited strong activity against ROS  
(reactive oxygen specie) production. The results are shown in (Table 4& Figure 5).  
Table. 4. The Anti-inflammatory activity in medicinal plant Sophora alopecuroides was determined in  
methanolic extract.  
Concentration  
Methanolic extract  
Ibuprofen  
% inhibition  
78.04  
Conc( μg/ml/μm)  
250 μg/ml  
IC50± SD μg/ml  
56.14± 2.82  
11.2±1.9  
73.2  
25μg/ml  
Figure. 5. Comparison of anti-inflammatory activity of methanolic extract and ibuprofen.  
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Conclusion  
Conclusion  
The present study was comprised on the identification of volatile constituents and biological activities of medicinal  
plant Sophora alopecuroides.  
The results of GC-MS analysis revealed the presence of 34 volatile constituents. The compounds were belong to  
4.12%, fatty acid derivatives 8.45%, carbohydrates 21.46%, steroids 1.01%, terpenoids 3.13%, alkaloids 0.27%,  
oxidative property 2.51%, cycloalkanes 0.63%, phenolic compound 0.21%, sulphur containing compounds  
11.23%, ketones 0.66 %.  
The FTIR spectrum confirms the presence of alcohol, alkane, carbonyl compound and ester functional groups.  
The strong peak at 3267.78 cm⁻¹ showed an O-H stretching vibration, which is typical of an alcohol functional  
group. The peak at the position of 1634.45 cm⁻¹, which denote stretching of carbonyl group C=O. The presence  
of alkane was confirmed and it appeared at 1406.47cm⁻¹ and might be C-H bending vibrations. The peak appeared  
at 1009.41cm⁻¹, which denote stretching of RCOOR.  
The plant was subjected for antibacterial and anti inflammatory activities. The methanolic extract of the plant  
showed moderate antibacterial activity against gram positive and gram negative bacterial strains, Staphylococcus  
aureus and Escherichia coli respectively. The plant extract showed strong anti-inflammatory activity against  
standard ibuprofen.  
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It can be concluded that according to the literature review and present findings, this plant possess variety of the  
compounds and showed promising results for the biological activities. It is also suggested that it can be studied  
further for the isolation of potential bioactive compounds which can be assessed against different biological  
activities.  
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