Volume 10, Issue 7, July – 2025 International Journal of Innovative Science and Research Technology

ISSN No:-2456-2165 https://doi.org/10.38124/ijisrt/25jul885

Evaluation of the Hypoglycemic Potential of the

Ethanolic Bulb Extract of Allium Chinense G. Don
(Sibujing) on High-Glucose Induced
Hyperglycemic Swiss Albino Mice
Kline Melody P. Nietes1; Raha D, Abdulrauf 2; Michaela Krystelle G. Ong3;
Christian Caleb G. Sacedon4; Junin Gay L. Garay5; Roselle T. Remulta6;
Rezeile Dale V. Pancho7
1,2,3,4,5,6,7
Department of Pharmacy, Adventist Medical Center College, Iligan City,
Lanao Del Norte, Philippines

Publication Date: 2025/07/19

Abstract: This study investigated the hypoglycemic potential of the ethanolic bulb extract of Allium chinense G. Don,
commonly known as Sibujing, a spice used in Maranao cuisine. Given the rising global prevalence of hyperglycemia and
diabetes mellitus, with 14,416 deaths due to diabetes mellitus in the Philippines alone in 2023, effective treatments are crucial.
Swiss albino mice were induced with hyperglycemia via a high-glucose diet to evaluate the extract's potential in managing
elevated glucose levels. Phytochemical screening of the crude extract revealed the presence of flavonoids, alkaloids, phenols,
saponins, steroids, and terpenoids, with the absence of tannins. Quantitative analysis showed a total phenolic content (TPC)
of 13.2±1.0 mg gallic acid per gram extract and a total flavonoid content (TFC) of 9.8±0.1 mcg quercetin per gram extract.
Acute oral toxicity testing indicated an LD50 >2000 mg/kg body weight, with no observed deaths or toxic symptoms.
However, statistical analysis using One-Way ANOVA and post-hoc tests demonstrated that administration of the crude
extract at varying doses (50, 300, 2000 mg/kg) did not result in a significant reduction in blood glucose levels compared to
both the negative control (normal saline solution) and the positive control (metformin, 50 mg/kg). All pairwise comparisons
yielded non-significant results (p>0.05), suggesting that under the conditions of this study, the ethanolic bulb extract of A.
chinense did not exhibit significant hypoglycemic activity comparable to metformin.

Keywords: Hyperglycemia, High Glucose, Crude Extract, Allium Chinense G. Don, Swiss Albino Mice.

How to Cite: Kline Melody P. Nietes; Raha D, Abdulrauf; Michaela Krystelle G. Ong; Christian Caleb G. Sacedon; Junin Gay L.
Garay; Roselle T. Remulta; Rezeile Dale V. Pancho; (2025) Evaluation of the Hypoglycemic Potential of the Ethanolic Bulb
Extract of Allium Chinense G. Don (Sibujing) on High-Glucose Induced Hyperglycemic Swiss Albino Mice. International
Journal of Innovative Science and Research Technology, 10(7), 1254-1263. https://doi.org/10.38124/ijisrt/25jul885

I. INTRODUCTION Organization (WHO) reported 642 million people living with

DM globally, with 1.6 million deaths annually. In the
Diabetes Mellitus (DM), a chronic disease Philippines, diabetes mellitus was the fourth leading cause of
characterized by elevated blood glucose levels death in 2023, accounting for 6.2% (14,416) of total deaths
(hyperglycemia), is a significant global public health concern. (Philippine Statistics Authority, 2023). This escalating
Metabolic disorders leading to insufficient insulin production prevalence underscores the urgent need for effective and
or utilization affect millions worldwide. The World Health accessible treatments.

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ISSN No:-2456-2165 https://doi.org/10.38124/ijisrt/25jul885
Traditional medicine, particularly the use of medicinal presence of flavonoids, alkaloids, saponins, tannins, steroids,
plants, has gained increasing interest in diabetes management and terpenoids, based on procedures adapted from Tinoy et
due to the high cost and potential side effects of conventional al. (2024).
medications (Zanzabil, 2023; Rondilla et al., 2021). Over
80% of the world's population relies on traditional medicine  Flavonoids (Hydrochloric Acid Reduction Test): Defatted
for primary healthcare (WHO, 2023). Allium chinense G. extract dissolved in 80% alcohol, treated with 12M HCl,
Don, locally known as Sibujing, is a widely available plant and heated. Red color indicates positive.
species in the Philippines traditionally used as a condiment in  Alkaloids (Wagner’s Test): Extract treated with 2M HCl
Mindanao (Vera Cruz et al., 2023). While other Allium and NaCl, filtered, then reacted with Wagner’s reagent.
species like A. sativum (garlic) and A. cepa (onion) have Brown precipitate indicates positive.
established antidiabetic properties due to their sulfur-  Saponins (Froth Test Method): Extract shaken vigorously
containing compounds (Nakamura et al., 2021; Choi et al., with water. Stable froth (2 cm for 30 minutes) indicates
2022; Pandey et al., 2022), the medicinal properties of A. positive.
chinense G. Don remain insufficiently studied, particularly its  Tannins (Ferric Chloride Test): Extract boiled with water,
potential in managing Type 2 Diabetes Mellitus (T2DM). filtered, and treated with 1% FeCl3. Blue or black
precipitate indicates positive.
Sulfur compounds are essential for plant health and  Steroids (Salkowski Test): Defatted extract treated with
human disease prevention, imparting antioxidant, anti- FeCl3, then concentrated H2SO4 layered at the bottom.
inflammatory, and antimicrobial properties (Hill et al., 2022). Brown, blue, or green rings indicate positive.
The Allium family is known for thiosulfinates, which  Phenols (Sodium Hydroxide Method): Extract dissolved
contribute to maintaining glucose tolerance and beta-cell in water, treated with 20% NaOH. Blue color indicates
viability by reducing oxidative damage and improving insulin positive.
secretion (Sabiu et al., 2019). This study aimed to investigate
 Terpenoids (Liebermann-Burchard Test): Extract
the potential hypoglycemic activity of the ethanolic bulb dissolved in chloroform, treated with acetic anhydride and
extract of A. chinense G. Don in high-glucose induced
concentrated H2SO4. Dark green color indicates positive.
hyperglycemic Swiss albino mice. Specifically, the research
 Quantitative analysis for Total Phenolic Content (TPC)
sought to determine the extract's effect on blood glucose
and Total Flavonoid Content (TFC) was also conducted.
levels, compare its efficacy to a standard antidiabetic drug
 Total Phenolic Content (Folin-Ciocalteu Method): Extract
(metformin), and identify the secondary metabolites present.
The findings could contribute to the existing knowledge of A. reacted with Folin-Ciocalteu reagent and sodium
hydroxide. Absorbance measured at 760 nm or 765 nm
chinense G. Don and its potential as a natural remedy for
using a spectrophotometer. Results expressed as mg gallic
T2DM.
acid equivalent (GAE) per gram of extract.
II. MATERIALS AND METHODS  Total Flavonoid Content (Aluminum Chloride
Colorimetric Assay): Extract reacted with aluminum
 Plant Material and Extraction chloride and sodium acetate. Absorbance measured at 415
Bulbs of Allium chinense G. Don were collected from nm using a UV-vis spectrophotometer. Results expressed
Wato-Balindong, Lanao del Sur, Philippines. The plant as mcg quercetin equivalent (QE) per gram of extract.
material underwent authentication and phytochemical
analysis at the Biological Science Laboratory of Mindanao  Animal Housing and Ethical Considerations
State University-Iligan Institute of Technology (MSU-IIT). Adult Swiss albino mice were obtained from the AMCC
The bulbs were cleaned, air-dried, and pulverized into a fine animal house and acclimatized for 7 days in the AMCC
powder using a pulverizer and sieved through No. 40 and No. Pharmacy Laboratory at 25±2∘C and 70% humidity. They
60 sieves to ensure uniform particle size (Tinoy et al., 2024; were fed standard rodent pellets and water ad libitum. The
Veltkamp, 2024). Two hundred grams of the powdered bulb room maintained 10-20 air changes per hour with a 12-hour
were macerated in 1 liter of 90% ethanol (1:10 ratio) for one light/dark cycle. Cages with wood shavings were cleaned
week with intermittent agitation at 150 rpm using an orbital thrice weekly with sodium hypochlorite (OECD, 2001). All
shaker (Kumar et al., 2023; Salem et al., 2016). Ethanol was animal procedures adhered to ethical guidelines, and animals
chosen as the solvent due to its polarity, miscibility with were humanely euthanized via cervical dislocation at the
water, and safety profile compared to methanol (Abubakar et study's end (IACUC Guidelines, 2014).
al., 2020; Pohanka, 2016). After maceration, the crude extract
was concentrated via water bath evaporation (Tinoy et al.,  Acute Oral Toxicity Test
2023). The acute toxicity test followed OECD Guidelines 423.
Twelve healthy Swiss albino mice were used (3 mice per
The percentage yield was calculated using the formula: step). The crude extract was administered via oral gavage at
%w/w yield=(weight of crude extract in g/weight of dried starting doses of 5 mg/kg body weight, followed by 50 mg/kg,
bulbs in g)×100. 300 mg/kg, and 2000 mg/kg if no toxic symptoms were
observed. Animals were observed for 30 minutes, 4 hours, 24
 Phytochemical Analysis hours, and daily for 14 days for any toxic signs, reactions, or
Qualitative phytochemical screening was performed on delayed effects. Internal organs were inspected post-
the ethanolic bulb extract of A. chinense G. Don to detect the dissection for lesions or deformities. The LD50 was estimated
based on these observations.

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ISSN No:-2456-2165 https://doi.org/10.38124/ijisrt/25jul885
 Induction of Diabetes and Treatment Administration All test substances were administered via oral gavage
After acclimatization, mice were induced with (Zake et al., 2021). Blood glucose levels were monitored at
hyperglycemia by oral gavage of 20 g/kg of 50% D-glucose 15-minute intervals for the first 1 hour and 45 minutes post-
for 2 weeks (Zhan et al., 2019). Blood glucose levels were administration.
monitored weekly via tail pricking using a OneTouch Select
glucometer and test strips (OECD, 2001). Once 25 mice  Statistical Analysis
reached a blood glucose level of 240 mg/dL (Melior Data was analyzed using One-Way Analysis of
Discovery, 2020), they were randomly divided into 5 groups Variance (ANOVA) to compare baseline blood glucose levels
(n=5 per group) for treatment over an 8-week period: among groups and to assess the overall effect of treatment.
Repeated measures ANOVA were performed to evaluate
 Group 1 (Low Dose): 50 mg/kg A. chinense extract changes in blood glucose levels over time and the interaction
 Group 2 (Mid-Dose): 300 mg/kg A. chinense extract between time and treatment. Post-hoc comparisons were
 Group 3 (High Dose): 2000 mg/kg A. chinense extract conducted to identify specific group differences when a
 Group 4 (Positive Control): 50 mg/kg metformin significant overall effect was observed (Ostertagova &
(standard dose, Sahu et al., 2024) Ostertag, 2013; Libretexts, 2024). Statistical significance was
 Group 5 (Negative Control): 100 mL/kg normal saline set at p<0.05.
solution

III. RESULTS AND DISCUSSIONS

 Preparation of Plant Extract

The crude ethanolic bulb extract of Allium chinense G. Don was dark-brown, semi-solid in mass, and had a pungent odor.

Fig 1 Crude Ethanolic Extract of A. Chinense G. Don

The average percentage yield based on the three replicants of 20 grams of dried powdered bulbs of A. chinense G. Don soaked
in 200 ml of ethanol is 66.67%. Table 1 shows the individual yield obtained from each replicant.

Table 1 The percentage yield of the crude ethanolic bulb extract of A. chinense G. Don
Replicant Weight of dried leaves (g) Weight of crude extract (g) Percentage yield (%)
1 20 3.3 66
2 20 3.3 66
3 20 3.4 68
Mean Value 66.67

Both replicants 1 and 2 yielded 66% while the third replicant yielded 68% in every 20 grams of the crude extract hence, the
mean value is 66.67%.

 Phytochemical Analysis
The phytochemical screening for secondary metabolites of the ethanolic bulb extract of A. chinense G. Don followed the
procedures and tests found in chapter 3 to determine the presence of phytochemical compounds such as flavonoids, alkaloids,
saponins, tannins, and steroids (Tinoy et al. 2024). A quantitative analysis of the Total Phenolic Content (TPC) and Total Flavonoid
Content (TFC) of the crude extract was also performed to quantify the number of flavonoids and phenols present in the extract
which are some of the phytochemicals responsible for the hypoglycemic activity of the Allium family (Nakamura et al.) (Hill et al.).

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Table 2 shows the results for the tests conducted.

Table 2 Results for Phytochemical Tests for Secondary Metabolites of A. chinense G. Don

The phytochemical analysis of the ethanolic crude was performed to determine the concentrations present in the
extract of A. chinense G. Don as shown in Table 2 shows a ethanolic bulb extract that may contribute to its hypoglycemic
positive result for alkaloids, flavonoids, phenols, saponins, activity (Sabiu et al. 2019) (Nakamura et al. 2021)
steroids, terpenoids, and a negative result for tannins. (Chakraborty et al. 2022) (Hill and Foroozand et al. 2023).
Table 3 shows the results for the quantitative analysis of A.
Additionally, a quantitative analysis of the Total chinense G. Don.
Phenolic Content (TPC) and Total Flavonoid Content (TFC)

Table 3 Quantitative analysis of the phenols and flavonoids of A. chinense G. Don

Quantitative Test Visible Results
Total Phenolics 13.2±1.0 mg gallic acid per gram extract
Total Flavonoids 9.8±0.1 mcg Quercetin per gram extract

The TFC and TPC of the crude extract of A. chinense body weight) of the ethanolic bulb extract of A. chinense G.
G. Don showed that the TFC is 9.8±0.1 mcg Quercetin per Don. The low dose was 50 mg/kg for the low dose, 300 mg/kg
gram extract and the TPC is 13.2±1.0 mg gallic acid per gram for the mid dose, and 2000 mg/kg for the high dose. The
extract indicating that there is a sufficient amount that can intestines, spleen, kidney, and liver were also in good
potentially cause a hypoglycemic activity. condition. There were no lesions, perforations, or any
deformities of any kind nor any changes to the texture of the
 Acute Toxicity Tests internal organs of the mice. Therefore, the LD50 is estimated
No deaths were recorded after 24 hours of at LD50 >2000 mg/kg body weight in mice.
administration of the various doses (3, 50, 300, 2000 ml/kg

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 Statistical Analysis repeated measures ANOVA revealed a significant main effect
The first part of the data was analyzed using the One- of time, F(7,140)=18.404,p<0.001, suggesting that blood
Way Analysis of Variance (ANOVA) method because it glucose levels significantly changed over the course of the
involves comparing the antidiabetic effects of the ethanolic study.
bulb extract of A. chinense G. Don to the positive and
negative control in Swiss albino mice to determine any However, the interaction between time and treatment
significant differences between the two variables. was not significant, F(28,140)=0.970,p=0.515, indicating that
the patterns of change over time were similar across groups.
A one-way analysis of variance (ANOVA) was The between-subjects effect of treatment was also not
performed to assess baseline differences in blood glucose statistically significant, F(4,20)=2.03,p=0.128, suggesting
levels among high-glucose induced diabetic Swiss albino that administration of the ethanolic bulb extract of Allium
mice assigned to control, low dose, mid dose, high dose, and chinense G. Don at different doses did not result in significant
metformin-positive groups. The analysis indicated no differences in blood glucose levels compared to the control
significant differences at baseline, F(4,20)=0.335,p=0.848. A and metformin-positive groups.

 Comparison of Crude Extract and Positive Control IV. DISCUSSION

A repeated measures ANOVA was conducted to
compare the hypoglycemic activity of the ethanolic bulb The present study aimed to evaluate the hypoglycemic
extract of Allium chinense G. Don at varying doses with that potential of the ethanolic bulb extract of Allium chinense G.
of the standard antidiabetic agent, metformin, in diabetic Don in high-glucose induced hyperglycemic Swiss albino
Swiss albino mice. The between-subjects effect of treatment mice. While the phytochemical analysis confirmed the
was not statistically significant, F(4,20)=2.03,p=0.128, presence of several bioactive compounds, including
indicating no significant differences in overall blood glucose flavonoids, alkaloids, phenols, saponins, steroids, and
levels among the groups. Although post hoc pairwise terpenoids, which are often associated with antidiabetic
comparisons were performed for transparency, all properties in other plant species (Tran et al., 2020; Nakamura
comparisons yielded non-significant results (p>0.05), further et al., 2021; Chakraborty et al., 2022; Hill et al., 2023), the
confirming the absence of meaningful differences between experimental results did not demonstrate a significant
treatment groups, including between the extract doses and hypoglycemic effect.
metformin.
The quantitative analysis showed a TPC of 13.2±1.0 mg
These findings suggest that the ethanolic bulb extract GAE/g extract and a TFC of 9.8±0.1 mcg QE/g extract. These
did not exhibit hypoglycemic activity comparable to that of values indicate the presence of phenolic and flavonoid
metformin under the conditions of this study. Below is a bar compounds, which are known for their antioxidant and
graph showing the relationship between the blood glucose potential glucose-lowering activities. For instance, A. cepa
levels of the mice and various doses over time. (onion) has been shown to reduce blood glucose levels due to
compounds like quercetin and s-methylcysteine sulfoxide,

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which promote glucose uptake and enhance insulin sensitivity The study's findings highlight the importance of
(Chakraborty et al., 2022; Ngan Tran et al., 2020). rigorous scientific evaluation of traditional remedies. While
A. chinense G. Don possesses various phytochemicals, its
However, despite the presence of these phytochemicals direct hypoglycemic efficacy, as tested in this study, was not
in A. chinense G. Don, their concentration or specific statistically significant. This does not preclude other potential
composition might not be sufficient, or their mechanism of health benefits or different pharmacological activities.
action might differ from those in other Allium species to elicit
a significant hypoglycemic effect under the study conditions. V. CONCLUSION AND RECOMMENDATIONS

The acute toxicity test indicated that the ethanolic bulb  Summary of Findings
extract of A. chinense G. Don is safe up to a dose of 2000 The ethanolic bulb extract of A. chinense G. Don was
mg/kg body weight, with no observed mortality or adverse successfully prepared, yielding an average of 66.67%.
effects. This suggests a wide therapeutic window if any Phytochemical screening confirmed the presence of
efficacy were to be found. flavonoids, alkaloids, phenols, saponins, steroids, and
terpenoids, but not tannins. Quantitative analysis revealed
However, the core finding of the study, as revealed by TPC of 13.2±1.0 mg GAE/g extract and TFC of 9.8±0.1 mcg
the repeated measures ANOVA, is the lack of a statistically QE/g extract. The acute toxicity test indicated an LD50
significant difference in blood glucose reduction across the >2000 mg/kg body weight, confirming the extract's safety at
various doses of A. chinense extract, the negative control, and the tested doses. However, statistical analysis using repeated
the metformin-positive control. While blood glucose levels measures ANOVA showed no significant differences in
did change significantly over time in all groups (likely due to blood glucose levels among the various A. chinense extract
the acute glucose challenge and subsequent physiological doses, the negative control, and the metformin-positive
regulation), the extract did not demonstrate a distinct glucose- control groups. The interaction between time and treatment
lowering pattern or magnitude comparable to metformin, nor was also not significant, indicating similar patterns of glucose
did it significantly differ from the untreated hyperglycemic change across all groups.
control. This suggests that the extract, at the tested doses and
under the specific experimental conditions (high-glucose  Conclusion
induction, Swiss albino mice model, and duration of Based on the current findings, the ethanolic bulb extract
observation), did not exert a notable antidiabetic effect. of Allium chinense G. Don did not exhibit a statistically
significant hypoglycemic effect on high-glucose induced
This outcome contrasts with some related studies on hyperglycemic Swiss albino mice under the conditions of this
other Allium species, such as A. hookeri, A. sativum, and A. study. The presence of secondary metabolites such as
cepa, which have shown promising antidiabetic effects in flavonoids, saponins, alkaloids, phenols, terpenoids, and
animal models (Choi et al., 2022; Jalal et al., 2007; Xie et al., steroids, while confirmed, did not translate into a measurable
2023). The discrepancy could be attributed to several factors: reduction in blood glucose levels comparable to the standard
antidiabetic drug metformin. Therefore, the null hypothesis,
 Species-specific differences: While belonging to the same stating that the ethanolic bulb extract of A. chinense G. Don
genus, the phytochemical profiles and biological activities would have no significant effect on blood glucose levels
of A. chinense may vary significantly from other Allium compared to the control group treated with metformin, is
species. accepted.
 Extraction method: Although ethanol is a common
solvent, different extraction methods or solvents might  Recommendations
yield different concentrations or types of active Based on these findings, the researchers recommend the
compounds. following for future studies:
 Dosage and duration: The tested doses, while covering a
wide range, might not have been optimal, or the 8-week  Alternative Diabetic Models: Utilize established diabetic
treatment period might have been insufficient to observe mouse models such as Non-Obese Diabetic (NOD) mice,
a significant long-term effect or a more pronounced Akita mice, or db/db and ob/ob mice, which may more
reduction in chronic hyperglycemia. closely mimic human T2DM pathophysiology and offer
 Animal model: While Swiss albino mice are common, different sensitivities to potential therapeutic agents.
other diabetic mouse models (e.g., NOD, Akita, db/db,  Chemical Induction Methods: Consider chemical
ob/ob mice) or different induction methods (e.g., alloxan induction of diabetes using alloxan or streptozotocin
or streptozotocin) might respond differently to the extract. (STZ) via the intraperitoneal route, which can provide a
The high-glucose induction method might have created a more consistent and well-characterized diabetic state.
hyperglycemia model that was not sufficiently responsive  Alternative Positive Controls: Explore therapeutic
to the extract's potential subtle effects. alternatives for positive controls, such as Linaligliptin or
 Metabolic complexity: T2DM involves complex Glibenclamide, which represent different classes of
pathophysiological mechanisms, including insulin antidiabetic drugs and may provide a broader comparative
resistance and β-cell dysfunction (Galicia-Garcia et al., context.
2020). The extract might not effectively target these  Further Formulation Studies: Given the presence of
specific pathways in the induced model. saponins, investigate the formulation of a soap containing

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Volume 10, Issue 7, July – 2025 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165 https://doi.org/10.38124/ijisrt/25jul885
A. chinense G. Don extracts. This would require solubility [10]. Philippine Statistics Office: 2022 Causes of Deaths in
testing in various solvents, compatibility testing with the Philippines (Preliminary as of 31 May
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