Description of Ciplukan Toxicity ( Physalis angulata L.)

Introduction: Ciplukan ( Physalis angulata L.) is a plant used by Indonesian people as traditional medicine. Drug sanitization needs to be carried out to guarantee the use and availability of scientifically safe traditional medicines. This study aims to provide information and enrich knowledge about the safety of consuming ciplukan roots and stems. Methods: This study used 8 male mice as test animals, divided randomly into 4 treatment groups, namely mice treated with a dose of 0.56 mg/20 g body weight; mice treated with a dose of 5.6 mg/20 g body weight; mice treated with 56 mg/20 g body weight; and mice treated with 560 mg/20 g body weight. Treatment was given once and then observed for 24 hours to observe the number of deaths of the test animals. Then follow-up observations were carried out in 3 days on individuals who were still alive. Results: Within 24 hours all individuals at the treatment dose of 0.56 mg/20 g body weight survived, whereas all individuals at the treatment dose of 5.6; 56; and 560 mg/20 g body weight died. The observations on individuals treated at a dose of 0.56 mg/20 g body weight showed that the animals were in good condition, with sleeping and eating activities, moving a lot, having clean and nice fur, and not showing toxic symptoms such as disturbances in physical activity, impaired balance, and refusal to eat. Conclusions: The administration of the test extract below is less or equal to 0.56 mg/20 g body weight is relatively safe.

Even though traditional medicine is medicine that comes from nature, the assumption that traditional medicine is safe to use and avoids harmful effects is wrong.The use of traditional medicine must be able to consider various things such as the correct dose, the exact time and method of use, as well as the accuracy of information review, because the inappropriate use of traditional medicine can potentially increase various unwanted symptoms.Not only synthetic drugs, traditional medicines also have a considerable risk of causing organ damage. 26To find out the benefits of a plant as a medicine, it is necessary to do toxicity testing.Toxicity is the harmful effect of a chemical compound on target organs in living organisms. 27Toxicity test is a preliminary test that is attempted to determine the toxic effects and thresholds for the use of a plant as medicine. 28Toxicity test aims to observe the pharmacological activity of a compound that occurs in a short time after exposure or administration in certain doses.Toxicity testing has the principle that bioactive components are always toxic when given in high doses and becomes a drug if given at low doses or measured doses. 29sed on the duration of the test, toxicity testing is divided into three groups, namely acute, subchronic, and chronic toxicity testing.Acute toxicity testing aims to find toxic effects in a short time, this test is useful for initial assessment of toxicity, determining target organs, obtaining hazard information after acute exposure to a substance, and sensitivity of test animals. 30Subchronic toxicity test is a test to detect toxic effects that appear after administration of the test preparation with repeated doses, the test preparation is given to the test animals for 10% of the entire age of the animals.The principle of the subchronic toxicity test is that the test group of animals is given the test preparation every day at several dose levels for 90 days. 31Meanwhile, chronic toxicity is a test conducted to determine the ability of a compound to cause adverse health effects as a result of long-term drug exposure. 32Of the three toxicity tests, the acute toxicity test is a test that needs to be carried out before carrying out other toxicity tests, 33 through the toxicity test is expected to be able to provide an overview of the initial data that can be used as a basis in determining a safe dose to avoid the occurrence of toxic effects. 4The toxic effect in question can be in the form of a burning book, 34 decreased movement activity, 4 and the death of the test animals. 35ven the potential possessed by ciplukan roots and stems and their extensive use as medicine, it is necessary to test the acute toxicity of ciplukan roots and stems.It is hoped that the results of this study can provide information and enrich knowledge about the safety of the ciplukan roots and stems themselves.

Extraction process
The extraction of ciplukan roots and stems refers to the Harborne 36 method.A total of 460.98 g of ciplukan roots and stems were cleaned, then cut into small pieces, and dried to produce a dry weight of 70 g.After that, the dried samples were macerated at room temperature using 96% ethanol solvent.The maceration process was carried out three times by adding new ethanol for each repetition.Then, the filtrate from the roots and stems of ciplukan was concentrated using a vacuum evaporator and a yield of 10.3 g was obtained.

Preparation of test animals
The test animals used were male white mice (Mus musculus), weighing 20 g-30 g, healthy condition, with an age range of 6-8 weeks.Prior to the experiment, all test animals were acclimatized for 7 days by providing standard food and drink ad libitum.The testing procedure has received a statement letter from the Health Research Ethics Commission, Faculty of Health Sciences, Respati University, Yogyakarta No. 019.3/FIKES/PL/I/2021.

Determination of toxicity and observation of accompanying toxic symptoms
The implementation of the toxicity test refers to Weil. 37A total of 8 mice as test animals were divided into 4 groups, each group consisting of 2 mice.Each test animal was given ethanol extract of ciplukan roots and stems, the test preparations were given orally.Each test group was given a different test dose with a 10-fold dose interval, namely a dose of 0.56 mg/20 g body weight (marked on the head); dose 5.6 mg/20 g body weight (with dorsal marking); dose of 56 mg/20 g body weight (with markings on the tail); and a dose of 560 mg/20 g body weight (no signs).To distinguish individuals at each dose, a yellow mark is given on the body of the test animal, while to distinguish individuals in each dose group, a dot is given on the tail of the test animal.The test was carried out by counting the number of test animals that died within the first 24 hours after administration of the test preparation.This acute toxicity test was modified by adding observations on individuals who did not experience death for 3 days, namely at 24 to 96 hours.

RESULTS
The abundance of biodiversity in Indonesia is utilized by the community, one of which is as traditional medicine.Utilization of biodiversity in the form of traditional medicine is an alternative that is considered more economical, 38 and generally have lower side effects than synthetic drugs. 11Traditional medicine will be safe and useful if it is used properly, be it the dosage, the duration of administration, the method of use, the selection of ingredients, and adjustments to certain indications. 39,40However, in the development of traditional medicines, inaccuracies in their use are often encountered and this occurs because of misinformation and erroneous assumptions about traditional medicines and how to use them.One of them is that synthetic drugs cannot be consumed in arbitrary quantities, traditional medicines also have dosages that must be adhered to. 26Therefore, many studies have been conducted to seek and collect scientific evidence about the safety and efficacy of traditional medicines.With scientific evidence, people can use traditional medicine safely and usefully. 41To test the safety of a traditional medicine, a toxicity test is carried out.The purpose of the toxicity test is to be able to assess various clinical symptoms and toxic effects caused by consuming plants used as traditional medicines 28,42 and ciplukan is one of the plants that has been traditionally used by the people of Indonesia, especially the people of West Kalimantan to treat jaundice and ciplukan is one of the plants that has been traditionally used by the people of Indonesia, especially the people of West Kalimantan to treat jaundice. 3compound contained in traditional medicine will be toxic if given at high doses and becomes a drug if given at low doses or measured doses. 29this means that a drug at a certain dose has the potential to be toxic in the body. 42Acute toxicity test is a toxicity test to look for the toxic effects of a chemical compound.The acute toxicity test is carried out by giving the test preparation once or several times within 24 hours. 42The acute toxicity test is a preclinical test designed to determine the lethal dose of a test preparation. 5Through the acute toxicity test it is expected to be able to provide an overview of damage to an organ as a result of the compounds contained in the test preparation, 4,43 the emergence of unwanted effects is influenced by many factors, including the type of substance involved, the size of the dose given, and the length of exposure to the substance in the body. 43Therefore, this study was conducted to evaluate the acute toxicity of ethanol extract of ciplukan roots and stems.The results of the acute toxicity test with graded doses for 24 hours are presented in table 1.An overview of the condition of the test animals during the administration of the test extract for 24 hours is presented in table 2.An overview of the conditions during the administration of the test extract from the 24th to the 96th hour is presented in table 3.

No Date Information Picture
One animal tested at a dose of 0.56 mg/20 g body weight moved and was agile, while the other animal tended to be quiet and sleep.
All animals tested at a dose of 5.6 mg/20 g, one animal treated at a dose of 56 mg/20 g body weight, and one animal tested at a dose of 560 mg/20 g body weight tended to be quiet and sleep, no activity.

DISCUSSSION
Each test animal was given ethanol extract of ciplukan roots and stems orally and in each test group a different dose was given at a 10-fold interval, namely a dose of 0.56 mg/20 g body weight (marked on the head); 5.6 mg/20 g body weight (with dorsal marking); 56 mg/20 g body weight (with markings on tail); and 560 mg/20 g body weight (no sign).The test was carried out by counting the number of deaths of the test animals during the first 24 hours after being given the test preparation, then followed by observations for 3 consecutive days on the test animals that were still alive.From the observations made it was known that at 06.45 in the morning the researchers gave ethanol extract of ciplukan roots and stems to the test animals and it was found at 08.02 in the morning there was death in 1 test animal treated with a dose of 56 mg/20 g body weight and 1 test animal treated with a dose of 560 mg /20 g of body weight, then at 14.30 noon there were also deaths in all test animals treated with a dose of 5.6 mg/20 g body weight, and additional deaths in 1 tested animal treated with 56 mg/20 g body weight, and 1 animal tested treated with a dose 560 mg/20 g body weight.Traditional medicine has a considerable risk of causing organ damage, the use of traditional medicine must consider the correct dosage, timeliness and method of use, as well as the accuracy of information review to prevent various unwanted symptoms. 26The death of mice showed that the increase in the dose given to the test animals and the length of observation time resulted in an increase in the percentage of deaths in the test animals, 44 Emilia et al. 45 stated that the high doses applied to the test animals would result in the test animals being exposed to the most of the tested extracts, and the length of time of observation of the test animals would give an illustration that initially the test animals were able to adapt to the doses given then gradually weakened because of the test extracts.inside his body begins to react and accumulate which will cause toxic symptoms.Toxic symptoms observed before the test animals died were individuals who were agile, moved a lot, with sparse fur, slept, moved little, and died.Clinical effects of adverse toxicity tests are effects that may produce functional impairment of organs and biochemical lesions that may alter the function of the organism in general or of specific organs. 46urthermore, Fithria et al. 34 states that clinical symptoms from toxicity testing can include changes in behavior, changes in skin and coat color, hair loss, gastrointestinal (constipation and diarrhea), vasodilatation, and movement (violent, tremor, convulsions, paralysis, and passivity), and death.
According to Aufia et al. 47 toxicity tests are used to observe the pharmacological activity of a compound that has bioactive components that are toxic when given at high doses.The existence of a toxicity test can provide an overview of initial data in determining a safe dose and to avoid toxic effects. 48Furthermore, the acute toxicity test will provide information about the lethal dose, therapeutic index, and degree of safety of the tested agent. 46Information on the lethal dose of a test compound can be seen from the number of mice that died and mice that were still alive in the test group, 48 this corresponds to Chinedu et al. 46 stated that the main parameter for measuring acute toxicity is to assess the dose that can kill 50% of the test animal population.In this study, the death of the test animals was related to the effects that appeared after administering graded doses of the compounds contained in ciplukan root and stem extracts.In ciplukan roots there are alkaloid group compounds, 10 phenolics, 49 alkaloid, phenolics, flavonoid, anthraquinone, and terpenoid. 50These studies show that ciplukan roots contain alkaloid compounds, 10,50 phenolics, 49,50 flavonoid, anthraquinone, and terpenoid. 50The ciplukan stems are reported to contain steroid group compounds, alkaloid, saponin, and flavonoid, 9 phenolics and flavonoid, 49 alkaloid, steroid, flavonoid, anthraquinone, and terpenoid. 50From these studies it can be seen that ciplukan stems contain alkaloid compounds, flavonoids, 9,49,50 steroid, 9,50 saponin, 9 phenolics, 49 and anthraquinone. 50en though these compounds are useful in the body, in reality the use of chemical compounds must be within the specified limit because if it exceeds the limit it will cause toxic effects on the body.Alkaloid compounds that have a harmful impact on the body are caffeine which causes dependence, 51,52 abnormal hearbeat, 51 tremors, 51,53 anxiety, 54 headaches, insomnia, stomach and digestive disorders, anxiety, and memory loss. 51Compounds of the saponin group have negative effects on livestock, which can irritate the lining of the mouth and digestive tract, affect nutrient absorption, fail to form zygotes, fail implantation, and abortion, and reduce iron absorption. 55Steroid compounds also have harmful effects on the body, namely female masculinization, testicular atrophy, gynecomastia in males, stretch marks, tissue damage, nerve injuries, paralysis, behavioral and personality disorders, and even death. 56Meanwhile, according to Widyasari, 57 compounds belonging to the flavonoid group can cause poisoning effects on the body and the resulting effects are allergies.Anthraquinone compounds can cause digestive discomfort, stomach cramps, dermatitis, nausea, vomiting, dizziness, and bloody diarrhea. 58The phenolic compounds have the effect of inactivating enzymes and denaturing proteins which causes disruption of the transport of organic ions into cells, thereby inhibiting cell growth or even causing cell death. 59xic symptoms appeared in the treatment group at doses of 5.6 mg/20 g body weight, 56 mg/20 g body weight, and 560 mg/20 g body weight.Before the test animals died, the test animals tended to be less mobile, and bristling/sparse fur was found in the test animals treated at a dose of 5.6 mg/g body weight.The decrease in movement activity in test animals is closely related to depression of the central nervous system and muscle relaxation, 4 whereas bristling hair is a general sign of toxicity which affects the integrity of the coat, in this case the active compounds present in the test material cause hair loss so that the hair on the test animal looks sparse. 34According to Yusuf 35 the death of

Table 1 : The number of deaths of test animals due to administration of ethanol extract of ciplukan roots and stems. Observation of death was carried out for 24 hours after administration of the test preparation with graded doses.
Panjaitan RGP, et al.: Description of Ciplukan Toxicity (Physalis angulata L.)