The Acute Toxicity of Ki Hampelas Leaves ( Sterculia rubiginosa Zoll . Ex Miq )

Sterculia is a genus with many pharmacological activities and is also use as a material in pharmaceutical preparations. Among the genus, Sterculia, Sterculia villosa Roxb, has antileishmanial activity and a safety level at giving a non-toxic dose of 100 mg/kg body1. Sterculia setigera Del. has the activity of the ethanol extract of Sterculia setigera Del. has antioxidant activity with the DPPH method on the fruit of 91.81%. In contrast, ethanol extract on stem bark is 86%2. Seeds of Sterculia foetida seed have antioxidant and antimicrobial activity3. Sterculia setigera Del., has activity as a tyrosinase inhibitory activity. It also has antiproliferative activity against human colon adenocarcinoma HT294. Sterulia quinqueloba (Garcke) K. Schum has activity as an antimycobacterial activity5. Sterculia quadrifida R.br Stem bark also has activity as an anti-hepatitis C6. Plants of the genus Sterculia are also using in n drug delivery applications7. Ag nanoparticles using seeds from Sterculia foetida L. have activity on mosquito vectors and HeLa cancer cells8. Some Sterculia genus plants have activities. Sterculia foetida for antibacterial and hemolytic10, apoptosis11, Sterculia diversifolia for immunomodulatory and anti-cancer12, Sterculia villosa as fibrinolytic5, sedative6, Sterculia tragacantha as anti-inflammatory and analgesic15,


INTRODUCTION
Sterculia is a genus with many pharmacological activities and is also use as a material in pharmaceutical preparations. Among the genus, Sterculia, Sterculia villosa Roxb, has antileishmanial activity and a safety level at giving a non-toxic dose of 100 mg/kg body 1 . Sterculia setigera Del. has the activity of the ethanol extract of Sterculia setigera Del. has antioxidant activity with the DPPH method on the fruit of 91.81%. In contrast, ethanol extract on stem bark is 86% 2 . Seeds of Sterculia foetida seed have antioxidant and antimicrobial activity 3 . Sterculia setigera Del., has activity as a tyrosinase inhibitory activity. It also has antiproliferative activity against human colon adenocarcinoma HT29 4 . Sterulia quinqueloba (Garcke) K. Schum has activity as an antimycobacterial activity 5 . Sterculia quadrifida R.br Stem bark also has activity as an anti-hepatitis C 6 . Plants of the genus Sterculia are also using in n drug delivery applications 7 . Ag nanoparticles using seeds from Sterculia foetida L. have activity on mosquito vectors and HeLa cancer cells 8 . Some Sterculia genus plants have activities. Sterculia foetida for antibacterial and hemolytic 10 , apoptosis 11 , Sterculia diversifolia for immunomodulatory and anti-cancer 12 , Sterculia villosa as fibrinolytic 5 , sedative 6 , Sterculia tragacantha as anti-inflammatory and analgesic 15 , One of the plants used as an ingredient in traditional medicine is Ki Hampelas leaves (Sterculia rubiginosa Zoll. Ex Miq). These plants are scattered in tropical and subtropical areas, especially in Sumatra. Based on previous research, Ki Hampelas leaves contain tannins, flavonoids, alkaloids, steroids-terpenoids, glycosides, and phenols. The antioxidant activity and total flavonoids equivalent to quercetine 17 . So it is interesting to study whether Sterculia rubiginosa has antioxidant and nephroprotective activity 16 .
To ensure the safety, effectiveness, and quality of a drug must undergo a series of tests. Starting from screening to look for active compounds, then continued with testing the effectiveness or selectivity and its mechanism of action in experimental animals or microbes, after being declared to have certain pharmacological activities by a series of safety tests on experimental animals, the toxicity test 17 . In previous studies, Ki Hampelas leaves have activity as a nephroprotective and antioxidant with a minimum dose of 50 mg/kg 16 . So it is necessary to do a toxicity test to ensure the safety of Ki Hampelas leaves extract.
The acute toxicity test aims to detect toxic effects that appear quickly and determine the LD 50 value in a compound or substance after being given a single dose or repeated doses within 24 hours.

Material
The leaves of Ki Hampelas obtained from the Bogor Botanical Gardens, Indonesia. Determination at the Botanical Garden Plant Conservation Center, Botany Division, Biology Research Center-LIPI, Indonesia. Ethanol obtained from local suppliers. Urea reagent kit, Creatinine kit, SGOT kit, SGPT kit from PT. Human, Indonesia. Na CMC (PT. Brataco), aqua destillata, obtained from local suppliers.

Extraction
Ki Hampelas leaves powder was extracted by maceration method with 70% ethanol. Two hundred (200) grams of Simplicia powder by immersing 70% ethanol. Then concentrated using a vacuum rotary evaporator at a temperature of 50ºC to obtain a thick extract, evaporated on a water bath with a temperature of 50ºC until a viscous extract 18 .

Extract characteristics
The extract was indentified with organoleptic, the yield, moisture content, ash content and phytochemical screening.

Acute toxicity test
Acute toxicity test used white male rats Sprague Dawley strain. The tested animals used in this study were 24 animals divided into four groups consisting of 6 animals for each group. The normal control group was given standard feed and 0.5% Na CMC, extract dose of 50 mg/kg, 1000 mg/kg, 2000 mg/kg. The test animals acclimatized for seven days. During acclimatization, the rats were fed and drunk and controlled the health and weight. Before taking blood, the animal test has fasted for ± 12 hours. The animals anesthetized using ketamine intramuscularly, then blood serum samples taken from white rats. The liver's acute toxicity parameters determined by SGOT, SGPT, and histopathology of the central vein and pyknotic nucleus size. The kidneys' observation done by determining creatinine, urea, and histopathology by looking at the cast on the tubules and the percentage of tubules with close and glomerular swelling. Glomerular calculated by measuring the distance from the Bowman's capsule's edge to the glomerular rim 19 . Tubular damage calculated using the formula = (n / m × 100%), where n= is the number of proximal tubules covered in one field of view and m= is the number of all proximal tubules in one field of view. Then the results were averaged to get the percentage of kidney damage in each mouse 20 . This research was permitted by the Ethics Committee KEPK-UHAMKA No. 02 / 20.03 / 0358.

Data analysis
The Kolmogorov-Smirnov and Levene's test determined the homogeneity and normal distribution of the data. The analytical continuous with the Tukey Test.

Phytochemical screening
Flavonoids, glycosides, alkaloids, tannins, saponins, triterpenoids and steroids, are present in the extract. The test results show in Table 2.

Acute toxicity of Ki hampelas leaf extract
The results Ki Hampelas leaf extract with the highest dose of 2000 mg/ kg did not cause death in the test animals, the value of LD 50 extract of Ki Hampelas leaves for test animals male white rats Sprague-Dawley is more than 2000 mg/kg.

Heart
At SGOT levels, SGPT showed an increase between the normal group and the extract group. However, this increase is still in the normal range, so it concluded that Ki Hampelas leaves extract's giving does not influence it. From the results of examining SGOT and SGPT levels, the value obtained (p> 0.05) indicated no significant difference in each treatment group. The results show in Figures 1 and 2.

Liver histopathology
Histopathological observations of the liver include measuring the central vein's diameter and the number of pyknotic nuclei. The central vein's diameter is used in the measurement because the central venous area is the center of the hepatic lobule and part of the reservoir of blood originating from the hepatic artery and portal vein 21 . The results of liver histopathological preparations show in Figure 5.
The parameter of liver organ, the central vein's diameter, between normal and the test group, shows no difference between groups. This result show in figure 6. The pyknotic nuclei show the differences between the normal and the test groups. Besides, the number of pyknotic nuclei which a sign of the occurrence of cell necrosis. This result show in Figure 7.

Kidney
The creatinine and urea levels of rats given the extract showed that treatment with different doses had increased creatinine and urea levels in rats. However, the increase in levels was still within normal limits. The results showed that creatinine and urea levels found that acute toxicity Ki Hampelas ki leaf extract had no significant effect on creatinine and urea levels p> 0.05. The results show in figures 3 and 4.

Kidney histopathology
Observations on the structure of the kidney structure include the distance between the glomerulus and Bowman's capsule, the percentage of tubules that close in one view and the cast on the tubules, can be seen in Figure 8. Casts are a collection of proteins that result in channeling through the tubular renal hampered, also stimulate the occurrence of necrosis of the tubules. While necrosis is death Jarin gan due to the absence of metabolites 22 .
The result of the study showed no presence of cast in control normal and the test groups. The percentage of tubules that closed in one field of view showed that there was no significant difference between the normal group and the 50 mg/kg dose group p>0.05, and the results between the 1000 mg/kg dose group and the 2000 mg/kg dose test group not seen. The significant difference with p> 0.05 shown in Figure 9. Meanwhile, the bowman room distance shows a significant difference between the normal control and test groups. The result show in Figure 10.

CONCLUSION
The ethanol extract of 70% Ki Hampelas leaves in the acute toxicity test did not cause death and toxic effects. The examination of SGOT and SGPT levels showed no significant difference between the dose treatment group and the normal group (p> 0.05).

ETHICAL ISSUES
The Ethics Committee permitted this research with the number KEPK-UHAMKA No. 02 / 20.03 / 0358.