INTRODUCTION:

Recently, the fruit of Hylocereuspolyrhizus, known as Red Dragon Fruit (RDF), has received a lot of attention from farmers around the world.¹In addition to the pulp, the skin of dragon fruit is also useful as an herbal medicinal materialand contains higher antioxidants than the pulp.²Antioxidants can be beneficial to neutralize elevated free radicals, protect cells from toxic effects and contribute to the prevention of various diseases.^3,4

When conventional pharmaceuticals are unable to effectively cure an illness, herbal therapies may be used as preventative measures, controls, or treatments for a variety of health issues. Concerns regarding the effectiveness and safety of phytopharmaceuticals have existed for a long time. The scientific community was also concerned about phytopharmaceuticals' potential side effects, toxicity, and medication interactions. For whatever cause, the safety of the herbal product or a certain dosage of a specific herbal component needs to be guaranteed.^5–7Tests that are usually carried out are toxicity tests which include acute, subacute, subchronic and chronic toxicity tests. Until now, there is little information on the safety of repeated exposure to dragon fruit peel extracts, so subchronic toxicity tests need to be carried out.⁸Toxicity studies are very important to identify the tolerance limit of extracts. Research by Kamatchi and Ravichelvan (2021) who evaluated the toxicity of H. undatus pulp in acute toxicity studies with various concentrations (100, 250, 500, 1000 and 2000mg/L) obtained an LD₅₀value of 446.68mg/L.¹ Research by Rizkifani et al. (2023) who conducted an acute toxicity test of red dragon fruit peel extract obtained an LD₅₀ value of more than 5000mg/kgWB and no deaths occurred during the acute toxicity test, which means that RDF peel extract does not provide acute toxic effects, then the safety of the extract needs to be further confirmed by repeated administration of the extract over a longer period of time through subchronic toxicity tests.³The LD₅₀ value is the dose that can statistically kill 50% of the experimental animals and the value is used as the basis for conducting subchronic toxicity tests.⁹

Subchronic toxicity study is a test to detect toxic effects that appear after administration of test preparations with repeated doses given orally to test animals during part of the animal’s life, but not more than 10% of the entire animal’s life.¹⁰The importance of subchronic toxicity testing is especially important for the use of traditional medicines or medicinal plants that are often used for a long time, in order to determine the spectrum of toxic effects.¹¹This test aims to determine the dose that does not cause toxic effects (no observed adverse effect level/NOAEL), to detect the toxic effects of the substance after repeated administration, and to study the cumulative effects and reversibility effects.¹²The principle of the subchronic toxicity test is that the test preparation in several dose levels is administered daily to several groups of test animals at one dose per group for 28 or 90 days, if necessary, satellite groups are added to see if there are delayed effects or reversible effects. During the time of administration of the test preparation, the animals should be observed daily to determine any toxicity. At the end of the test period, all surviving animals are autopsied and macropathological observations are made on each organ and tissue. In addition, hematological, clinical biochemical and histopathological examinations are also carried out.¹⁰Based on the descriptions above, researchers are interested in conducting a study on the subchronic toxicity test of RDFpeel extract.

MATERIALS AND METHODS:

Methods:

Sample Collection and Processing:

Sample were obtained from Sarang Burung Kolam Village, MatangTangkit Hamlet, Jawai District, Sambas Regency, and West Kalimantan Province. We gathered the peel of RDF while it was still fresh, clean, red, and undamaged. RDF peel sample had beewere then dried in the oven to a temperature of ±50°C until it was breakable without becoming too rigid. then ground 500 mg of dried simplicia. To mitigate the risk of impurities and preserve the simplicia powder, we kept it in a sterile and dry receptacle.

Preparation of Extract:

Extraction was done through maceration method. The simplisia powder was soaked using methanol solvent while occasionally stirring. The macerate was filtered every 24 hours and the residue was macerated again. This process was carried out for 3 days. The obtained macerate was concentrated using a rotary evaporator until a thick extract was obtained.

Approval from Ethics Committee:

The research was carried out after obtaining approval from the Ethics Committee of the Faculty of Medicine, Tanjungpura University with approval number 4452/UN22.9/PG/2023.

Experimental Animal:

The test animals used had the following inclusion criteria: non-pregnant; 2-3 months in age; and 100–200 grams of body weight. The exclusion criteria consisted of rats exhibiting physical abnormalities or defects. We kept the experimental room in pristine condition every day. In accordance with the OECD rule on number 407 (Organization for Economic Co-operation and Development), the room temperature was maintained within the range of 22.3oC, the relative humidity was maintained between 50 and 60%, and the lighting cycle consists of 12 hours of light and 12 hours of darkness. Six male and six female groups were formed by dividing the test animals using a random sampling method. Picric acid facilitated the differentiation of individual test animals by assigning numbers to each one.

Table 1. Animals Experimental Group

Group	Treatments
Control	Given food and drink (ad libitum)
Under dose	Given a dose of 1250 mg/kgWBRDFpeel extract
Middle dose	Given a dose of 2500 mg/kgWBRDF peel extract
Upper dose	Given a dose of 5000 mg/kgWBRDF peel extract
Control Satellite	Given food and drink (ad libitum)
Upper Dose Satellite	Given a dose of 5000 mg/kgWBRDFpeel extract

Subchronic toxicity Study^10,13

In this investigation, subchronic toxicity was assessed using the following parameters: body weight, behavior, organ index, biochemical and hematology contents of blood sample. Observations of body weight were conducted on day-1, day-7, day-14, day-28, and day-42 for the satellite group. The animal's behavior and motor activity were assessed at the following time points: 0 hours (prior to test preparation administration), 30 minutes, 1 hour, 2 hours, 24hours, and prior to its termination. Observations of the organ index were conducted subsequent to termination.

The OECD 407 Limit Test method was employed to conduct the subchronic toxicity test in this investigation. The limit test was conducted using three different dosing stages: 1250mg/kgWB, 2500mg/kgWB, and 5000 mg/kgWB. The extract was administered to the rat once a day. As the rat remained alive after 24hours, observations were maintained for 28 days and 42 days for the satellite group. The 42-day observation period was designed to detect any delayed toxic effects that were not apparent within the initial 28 day. The dose group test animals were given the extract orally every day for 28 days. After 28 days, the satellite group was left for 14 days without being given the extract. Rats were euthanised through chloroform inhalation and blood was taken directly from the heart using a 1 cc syringe on day 29 (for the dose and control groups) and 43 days (for the satellite group).

Organ Index:

The following organs were examined: spleen, heart, lungs, liver, and kidneys. Evaluations were conducted to determine whether or not each organ was damaged. Prior to obtaining their weight, organs must be dried using absorbent paper. Subsequently, the weight of the organ must be determined. The internal organ index percentage was calculated by dividing the weight of the organ by the weight of the test animal's body. The formula for calculating the organ index is as follows (BPOM, 2014):

Organ weight° (gram)

Organ Index = ------------------------------------ ×100%

Rat Body weight (gram)

Analysis Data:

The testing yielded both qualitative and quantitative information. The qualitative data were acquired through an examination of the motor activity and behavior of the test animals, as well as an assessment of the occurrence of fatalities or toxic effects. Body weight, behavior, organ index, biochemical and hematology contents of blood sample values were analyzed. The statistical analysis involved comparing the body weight and organ index of male and female rats using the Independent T-test with a 95% confidence level. To assess differences in body weight and organ index between subchronic toxicity test dose groups, a one-way ANOVA test was employed.

RESULT:

Body Weight Profile and Clinical Condition:

Throughout the 28-day treatment period and the recovery phase, there were no deaths or signs of straub, piloerection, ptosis, lacrimation, catalepsy, salivation, vocalization, tremors, convulsions, or writhing. normal observed behaviour during the research period, namely: respiration, feces, urine, hanging, establishment, flexion, haffner, and corneal and pineal reflexes. There was no difference in motor activity between the test and control groups.There was a decrease in motor activity in the satellite group, but it returned to normal after 14 days.An animal is most likely adjusting to its present circumstances if there are no symptoms of illness or unusual behavior in addition to good indicators of health and behavior.⁵

During the test, the animal's body weight was measured daily. According to the graph in Figure 1, weight loss occurred in the first week of the test period, especially in the first week of the test period. This decrease is considered temporary, as evidenced by the average body weight of the test animals which tends to remain stable.

Figure 1. Body weight profile after oral administration of RDFpeels methanol extract.

Organ Index:

The graph in Figure 2 shows the variation of organ index in each test group. However, this variation does not indicate a statistically significant difference.Figure 2 also shows noteworthy observations in male (A) and female (B) satellite groups, comprising the control group and the 5000mg/kg high dose group. A substantial increase in liver organ index was observed, especially at the 5000mg/kg dose. However, statistical tests showed no overall significant differences in this parameter.

Biochemical Profile:

The table below displays the rats' 28-day biochemical profile. It is known from the results that the levels of ALT, AST, and LDL increased in all groups of male and female rats. In male rats, the dosage group administered at 5000 mg/kgWB had highest elevation. In female rats, the 2500 mg/kgWB dosage group and the control group both had elevated LDL levels. Other parameters in the blood biochemical profile of test rats in this study showed levels at normal levels.

Transaminase Enzymes Profile:

Methanol extract of RDF peel caused an increase in AST levels in rats with the highest increase in the female control satellite group. Based on the results of One-Way Anova statistical analysis, there was a significant difference in AST levels between groups in male rats (p<0.05). (Table-3).

Figure 2. Index organ of male (A) and female (B) of each organ (Right and Left Kidney; Spleen; Liver; Heart and Lung) after oral administration of RDFpeels methanol extract.

Table 2. Biochemical Profile of RDFpeels methanol extract on rats.

Parameter	Male
Parameter	Control	1250 mg/kgWB	2500 mg/kgWB	5000 mg/kgWB	Reference Value
Glucose (mg/dL)	149.8425±15.6	170.265±61.0	116.865±36.9	106.545±29.5	100-200
TC(mg/dL)	90.335±5.0	98.595±3.8	89.2875±13.5	97.735±15.04	70-140
TG(mg/dL)	121.8275±12.1	119.995±13.0	107.7±18.6	107.425±5.72	50-200
LDL(mg/dL)	35.6325±8.12	38.605±6.53	37.5875±6.6	41.01±11.1	<27.20
Urea(mg/dL)	45.4425±9.09	50.03±3.9	43.9025±3.0	42.3775±4.41	10-59
Creatinine(mg/dL)	0.5125±0.09	0.6025±0.04	0.585±0.04	0.715±0.07	0.3-1.0
Parameter	Female
Parameter	Control	1250 mg/kgWB	2500 mg/kgWB	5000 mg/kgWB	Reference Value
Glucose (mg/dL)	174.61±38.0	110.375±25.1	145.67±50.5	113.6625±11.7	100-200
TC(mg/dL)	104.835±12.5	80.0325±25.2	87.77±16.4	68.675±10.5	70-140
TG(mg/dL)	150.2325±16.5	123.305±12.5	131.6425±16.4	132.485±5.8	50-200
LDL(mg/dL)	35.9775±6.64	27.3275±22.2	35.6275±17.9	25.6825±8.8	<27.20
Urea(mg/dL)	57.28±5.76	49.8175±0.5	50.64±10.1	49.095±3.5	10-59
Creatinine(mg/dL)	0.5325±0.129	0.5975±0.09	0.5425±0.09	0.435±0.13	0.3-1.0

Table 4. ALT enzyme profile of RDF peels methanol extract on rats.

Group	ALT Mean Value ± SD (U/L)
Group	Male	p Value	Female	p Value	Reference Value
Control	36.3725±3.17	0.123	80.1325± 81.4	0.245	10-50 U/L
1250 mg/kgWB	47.6925±13.64		53.835±33
2500 mg/kgWB	26.0325±6.98		60.6±42.4
5000 mg/kgWB	42.74±17.6		67.0725±32.1
Control Satelite	68.9525±54.1		96.05±87.8
5000 mg/kgWBSatelite	69.4675±13.14		56.36±41

Figure 4. ALT profile of rats after oral administration of RDFpeels methanol extract presented in bar diagram

Hematological Profile:

Table 5.Hematological Profile of RDF peels methanol extract on rats

Treatment	Parameters	Male			Female
Treatment	Parameters	Mean Value	SD	p-Value	Mean Value	SD	p-Value	Reference Value (Weiss &Wardrop, 2010)
Control	Haemoglobin (Hgb)	12.69	0.34	0.09	12.69	0.34	0.09	15.7 g/dl
1250 mg/Kg		13.93	1.77		12.7	0.37
2500 mg/Kg		12.27	0.58		12.71	1.71
5000 mg/Kg		12.77	0.51		11.98	1.65
Satellite Control		14.04	0.64		13.76	0.75
Satellite 5000 mg/Kg		13.1	0.49		13.45	0.66
Control	Leukocytes (WBCs)	7.37	0.5	0.45	7.37	0.5	0.45	4.52 × 10³/μL
1250 mg/Kg		8.35	2.17		9.05	3.71
2500 mg/Kg		8.45	1.69		7.25	2.91
5000 mg/Kg		7.5	1.83		8.3	2.2
Satellite Control		9.19	3.65		8.35	4.76
Satellite 5000 mg/Kg		7.8	3.28		10.29	0.84
Control	Erythrocytes (RBCs)	7.29	0.27	0.018	7.29	0.27	0.018	8.39 × 10⁶/μL
1250 mg/Kg		7.98	1.25		7.48	0.74
2500 mg/Kg		6.98	0.22		7.43	1.31
5000 mg/Kg		7.23	0.4		6.83	0.36
Satellite Control		8.45	0.58		7.95	0.88
Satellite 5000 mg/Kg		8.09	0.45		8.22	0.88
Control	Eosinophils	1.5	1.29	0.618	1.5	1.29	0.618	0.04 × 10³/μL
1250 mg/Kg		1.5	0.58		1.5	1
2500 mg/Kg		1.75	0.96		1.75	0.96
5000 mg/Kg		1.25	0.5		2	0
Satellite Control		1.5	1		1.25	0.5
Satellite 5000 mg/Kg		1.25	0.5		1.5	0.58
Control	Basophils	0	0	0	0	0	0	0.01 × 10³/μL
1250 mg/Kg		0	0		0	0
2500 mg/Kg		0	0		0	0
5000 mg/Kg		0	0		0	0
Satellite Control		0	0		0	0
Satellite 5000 mg/Kg		0	0		0	0
Control	Lymphocytes	69.25	8.62	0.102	69.25	8.62	0.102	1.56 × 10³/μL
1250 mg/Kg		70	17.81		76.25	5.56
2500 mg/Kg		64	9.56		68.5	13.63
5000 mg/Kg		66.5	3.42		71.25	4.57
Satellite Control		75.25	6.29		58.5	13.18
Satellite 5000 mg/Kg		49	8.6		55.5	7.72
Control	Monocytes	1.75	0.5	0.002	1.75	0.5	0.002	0.08 × 10³/μL
1250 mg/Kg		1.75	0.5		1.5	0.58
2500 mg/Kg		1.25	0.96		1	0
5000 mg/Kg		1.25	0.5		1.25	0.96
Satellite Control		1	1.15		1	0.82
Satellite 5000 mg/Kg		1.5	0.58		1.5	0.58
Control	Haematocrit (Hct)	37.48	0.21	0.083	37.48	0.21	0.083	45%
1250 mg/Kg		41.93	5.39		38.09	1.1
2500 mg/Kg		36.81	1.75		38.12	5.11
5000 mg/Kg		38.42	1.41		37.27	6.22
Satellite Control		42.12	1.93		40.94	2.27
Satellite 5000 mg/Kg		39.41	1.6		39.93	2.03
Control	Platelets	182.5	17.08	0.965	182.5	17.08	0.965	904 × 10³/μL
1250 mg/Kg		210	42.43		172.5	9.57
2500 mg/Kg		185	12.91		170	18.26
5000 mg/Kg		190	29.44		167.5	23.63
Satellite Control		192.5	9.57		170	18.26
Satellite 5000 mg/Kg		165	20.82		218.75	17.5
Control	MCV	54.46	5.45	0.018	54.46	5.45	0.018	53.5 fL
1250 mg/Kg		52.79	3.27		51.31	4.93
2500 mg/Kg		52.77	1.32		51.63	2.36
5000 mg/Kg		53.98	1.49		52.2	5.24
Satellite Control		49.95	2.18		51.58	3.31
Satellite 5000 mg/Kg		48.83	0.92		50.02	2.82
Control	MCH	17.41	0.97	0.657	17.41	0.97	0.657	18.7 pg
1250 mg/Kg		17.59	1.09		17.1	1.65
2500 mg/Kg		17.66	0.42		17.21	0.8
5000 mg/Kg		18.23	0.63		17.16	1.86
Satellite Control		16.94	0.56		17.25	1.16
Satellite 5000 mg/Kg		17.33	1.13		16.75	0.98
Control	MCHC	32.72	0.29	0.086	32.72	0.29	0.086	34.9 g/dl
1250 mg/Kg		34.02	1.69		32.64	0.68
2500 mg/Kg		32.36	0.47		32.79	1.69
5000 mg/Kg		32.86	0.51		32.01	1.64
Satellite Control		34.12	0.66		34.05	0.99
Satellite 5000 mg/Kg		33.42	0.75		33.34	0.78

DISCUSSION:

For thousands of years, supplementary and traditional medicine have been applied. Additionally, it is an evidence-based medication due to its extensive usage and popularity.¹⁴Herbal plants have traditional significance because they are not only important components of our food that have nutritional value but also have pharmacological properties, therefore this can provide starting materials for the synthesis of conventional drugs.^15,16Absence of rules and a fact-based approach are frequent and have detrimental effects. This might be because there are insufficient scientific study data and research methodologies to evaluate medications.^17,18Therefore, several herbs and their preparations have been identified for toxicity in order to ensure their safety around the world. To increase the market's acceptability, standardize, and/or regulate the sale of herbal medications, safety testing is required.¹⁷ This study was done to evaluate the sub-chronic toxicity of an extract obtained from the peel of RDF. The peel, originally discarded as food, has since been recognized for potential uses as a traditional medicine agent due to its antimicrobial, cosmeceutical, antioxidant, and food additive properties.^19–24

This study was conducted based on the OECD 405 method, in which toxicity studies were conducted for 28 days to evaluate the toxic potential of the extracts tested. Based on our previous study, single administration of RDF peel extract at doses of 1250 mg/kgWB, 2000 mg/kgWB and 5000 mg/kgWB showed no mortality or toxic symptoms in rats. After oral administration of the extract for 28 days, there was no mortality or abnormal behavior observed. Observations on the body weight of rats showed a decrease in body weight in all groups (Fig 1). However, after the administration of the extract was stopped, body weight tended to stabilise without any significant increase or decrease. The most significant weight loss was seen in the female rat group.Changes in body weight are a crucial indicator of an animal's health. The first need for the beginning of negative consequences is often weight loss. Toxic dosages are defined as those that alter body weight by 10% or more.²⁵

The organ index serves as a crucial parameter in subchronic toxicity assessments, facilitating the discernment of the impacts arising from repeated doses of RDFpeel extract administered over a 28-day period on the dimensions and functionality of vital organs in the employed test subjects, specifically Wistar strain white rats. The alterations observed in organs such as the liver, spleen, kidneys, lungs, and heart—both in terms of size and structural integrity—constitute a comprehensive evaluation, offering valuable insights into the in vivo safety profile of the tested substance.²⁶

Several organ indices in each dose group and satellite group exhibit significant values, indicating potential differences in size and organ damage. If there is an excessive significance in the liver organ index, it suggests morphological and physiological changes due to injury to liver cells, leading to increased water absorption spreading into the cell cytoplasm. Consequently, liver cells swell due to an increase in cell volume and size. However, due to the abundance of antioxidant compounds in herbs, many studies suggest that plant extracts are hepatoprotective rather than causing liver damage.The possible mechanism of herbal preparations as hepatoprotective agents is that they can significantly reduce lipid peroxidation through increasing MDA levels in liver homogenates.^27,28In the kidney organ index, an increase can occur due to inflammation, causing an enlargement of the kidneys. This is believed to result from the antihypertensive effects of potassium present in the combination of banana peel and pineapple peel extract, leading to a decrease in blood pressure to the kidneys and, subsequently, a reduction in blood flow to the kidneys. This condition can trigger hypoxia in renal tubule cells, potentially progressing to cell necrosis. Kidney cell necrosis is characterized by enlarged cell size due to inflammation, resulting in an increased kidney size.²⁹

The elevation in liver and kidney organ weight indices serves as an indicator of damage to the liver and kidneys. However, these organ index data cannot be considered an absolute parameter for assessing the impact of the test preparation on the organs. This is due to the possibility of annon-proportional relationship between body weight and the weight of the organs in the test animals.³⁰Comparisons are drawn between the organ indices of the control group and the dose group to meticulously gauge the repercussions of prolonged exposure to the test substance throughout the 28-day duration. Simultaneously, the organ indices of the satellite group are scrutinized with the intent of elucidating any reversibility or delayed effects ensuing from the test substance, a comprehensive examination conducted over a 14-day post-administration period subsequent to the initial 28-day exposure phase.²⁹

Biochemical assays showed no abnormalities in the enzyme ALT (alanine transaminase), AST (aspartate aminotransferase) and LDL (low-density lipoprotein)parameters.Transaminase levels in plasma are sensitive markers of damage to liver cells. Transaminase enzymes, such as ALT and AST, are mostly found in liver cells. When the liver is harmed, the liver cells release these enzymes into the bloodstream, increasing blood levels of ALT and AST.^7,31The increase in AST levels was seen in line with the increasing dose, this means that the higher the dose given, the greater the possibility of the increasement (Table 3). In contrast, there was a rise in the ALT parameter in practically every group of rats. Normal levels were seen in the male rat group at a dose of 2500 mg/Kg. Since the liver is primarily responsible for lipid production and metabolism, lipid profiles are markers that are used to detect problems with lipid metabolism control.All rat groups' total cholesterol and triglyceride levels fell within the normal range, according to the data collected. All groups did, however, see a rise in LDL levels above normal ranges, which can be brought on by excessive feed consumption. One indicator of metabolic problems, which might lead to fatty liver disease, is this elevated LDL level. This is consistent with the elevated ALT and AST values in rats, which suggest injury to the liver.^32,33

The hematological parameters, including Haemoglobin (Hgb), Leukocytes (WBCs), Erythrocytes (RBCs), Eosinophils, Basophils, Lymphocytes, Monocytes, Haematocrit (Hct), Platelets, Mean cell volume (MCV), Mean cell haemoglobin (MCH), and Mean cell haemoglobin concentration (MCHC), exhibit statistical significance at the 0.05 level. This significance is observed specifically in the parameters of Erythrocytes, Monocytes, and MCV following an analysis conducted using One Way ANOVA. These findings suggest anomalies associated with the administration of methanol extract doses of RDFpeel.

Alterations in erythrocyte count and haemoglobin levels can be attributed to environmental shifts, such as increasing temperatures, leading to fluid loss and resulting in morphological changes and haemoglobin release from erythrocytes.³⁴ Secondary metabolites, including alkaloids, tannins, saponins, and flavonoids, exhibit the capacity to elevate erythrocyte counts and reduce leukocytes.³⁵ Changes in monocyte count, a type of white blood cell, may signify potential issues such as inflammation, neoplasia, damage to progenitor or stromal cells, decreased blood output, or leukocyte adhesion defects, alongside pre-sacrifice stress in rodents.

MCV ranges from 45 to 55 fL in mice and 55.1 to 61.5 fL in rats. Mean corpuscular haemoglobin (MCH) lacks substantial utility due to its proportional variability with MCV. Mean corpuscular haemoglobin concentration (MCHC), when considered with MCV and red blood cell distribution width (RDW), proves invaluable for evaluating red blood cell changes in mice and rats. MCHC typically varies from 30 to 38 g/dL in mice and 30 to 34 g/dL in rats, contingent on species, gender, and age. RDW, indicating red blood cell size variation or anisocytosis, is ascertainable solely through automated haematology analyzers. Values may exhibit inconsistency across instruments due in part to differing formulas for RDW calculation.³⁶

CONCLUSION:

The subchronic toxicity test on Wistar white rats exposed to RDF peel extract indicated a favorable safety profile. Over 28 days of oral administration, no deaths or behavioral abnormalities were reported. Although significant weight loss occurred in female rats, it was the only notable adverse effect. Organ indices showed some significant values, particularly in the AST profile for male rats, with no significant differences in ALT profiles for both genders. Hematological profiles exhibited significant differences within acceptable limits, suggesting potential influences on blood parameters. Overall, repeated exposure to RDF peel extract at tested dosages (1250mg/kgWB, 2500 mg/kgWB, and 5000mg/kgWB) appeared generally safe. However, further research is needed to understand specific physiological mechanisms and long-term effects associated with the extract's consumption.

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

ACKNOWLEDGMENTS:

The authors would like to thank Zammi Laboratory for their kind support during hematological and all other lab studies.

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Received on 21.12.2023 Revised on 04.05.2024

Accepted on 02.09.2024 Published on 20.01.2025

Available online from January 27, 2025

Research J. Pharmacy and Technology. 2025;18(1):379-387.

DOI: 10.52711/0974-360X.2025.00059

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

Group	AST Mean Value ± SD (U/L)
Group	Male	p Value	Female	p Value	Reference value
Control	76.0775±2.25	0.075	129.4175±102.6	0.403	45-120 U/L
1250 mg/kgWB	85.0725±17.5		103.4±45.6
2500 mg/kgWB	67.56±7.24		116.2375±60.6
5000 mg/kgWB	90.1425±27.24		135.4625±42
Control Satellite	107.99±60		199.86±61.8
5000 mg/kgWB Satellite	127.7675±12.25		134.8075±53.5