Acute Toxicity Testing of White Turmeric Extract (Curcuma zedoaria) on Histopathological Imaging of the Lungs

White turmeric (Curcuma zedoaria) is a type of plant whose extract contains compounds that can inhibit carcinogenesis. Acute toxicity test was conducted to determine the safe dose and lethal dose (LD) 50 from the use of a drug substance. This research aimed to determine the effect of the acute toxicity test of white turmeric extract on the histopathological imaging of the lungs. This study is an experimental study with a post test only control group design. A total of 30 Wistar rats was divided into six groups. Data analysis was using one-way ANOVA statistical test, while for lung histopathology using ordinal data which were analyzed descriptively. In conclusion, the acute toxicity test of white turmeric extract on Wistar rats was not toxic and there was no death and no toxic symptoms and no necrosis, congestion and inflammation were found on the histopathological picture of the lungs.


Introduction
Healthy living is everyone's dream. However, this hope has been hampered by the increasing price of modern medicines and the possible side effects.
Nowadays, people tend to explore experiences from ancestral culture, as a back to nature trends. Furthermore, they start to use herbal medicine.
Traditional medicines and medicinal plants are widely used in the lower middle class, especially in preventive, promotive, and rehabilitative efforts. Meanwhile, many people think that the ingredients for using traditional medicines are relatively safer than synthetic drugs.
Generally, the efficacy of traditional medicines is only based on practical experience and has not been scientifically tested 2 .
White turmeric (Curcuma zedoaria) is one of the genus Curcuma has the characteristics of light green round leaves; flowers grow in clusters on pseudostems as high as 30-70 cm. 3 The shape of the rhizome is long like a finger, the outside parts are brownish-white, and the inside is yellowish-white. White turmeric rhizome contains the primary substance, namely curcumin, which contains diferuloylmethane as a natural antioxidant. Curcumin also has anti-inflammatory, anticancer, and antifungal functions. It is also known that white turmeric extracted has natural antioxidant

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Case Reports substances such as total phenols and total flavonoids and has antioxidant activity, namely free radical scavenging capacity and reactive oxygen species. 4 White turmeric extract contains compounds that can inhibit carcinogenesis. These compounds include essential oils, polysaccharides, and curcuminoids identified and isolated, including curcumin, dimethoxy curcumin, and bisdemethoxycurcumin. 5

Methods
The A total of 500 grams of white turmeric simplicia were extracted using the maceration method for 3 x 24 hours with 70% ethanol as solvent. Then, the macerate was evaporated using a rotary evaporator so that a thick extract was obtained for other weighing and preparation of the extract to be given to the rats. After acclimatization, rats were grouped into six groups (each group consisted of five rats): group 1 (rats were given aqua dest), group 2 (rats were given NaCMC), group 3 (rats were given white turmeric extract 250 mg/kg BW (WTE250)), group 4 (rats were given white turmeric extract 500 mg/kg BW (WTE500)), group 5(rats were given white turmeric extract 750 mg/kg BW (WTE750)), group 6 (rats were given white turmeric extract 2000 mg/kg BW (WTE2000)). The treatment was carried out for 14 days, where the treatment was given using an intragastric sonde.
Furthermore, clinical observations were carried out to assess the toxic symptoms of the treatment by assessing the rat skin, lethargy, diarrhea, tremor, and death. After 14 days, euthanasia was performed using ketamine 100 mg/kg BW intraperitoneally, and then the lungs were evacuated. Next, the lungs were put into a 10% Neutral Buffer Formalin solution (1:10). Then the dehydration process was carried out by gradually entering the lungs from 70% to 99%, followed by making paraffin blocks. Furthermore, the staining process was carried out with hematoxylin-eosin, and histopathological examination was carried out to assess changes and damage to lung tissue.
Data analysis was carried out using SPSS 25. The data were presented descriptively and narratively to explain the observations. Univariate and bivariate analyses were performed to assess differences in clinical symptoms and lung tissue damage, with p<0.05.

Results
Observations of toxic symptoms were carried out every day after being induced by white turmeric extract

Symptoms of toxicity after treatment
The toxicity test in this study used an extract of white turmeric (Curcuma zedoria) on Wistar rats (Rattus  that are not toxic, such as glucose or salt. If given in excess, these substances will damage the osmotic balance so that they can injure or cause cell death 17 .

Conclusion
Acute toxicity test of white turmeric extract (Curcuma zedoria) is not toxic and no death arise in rats. The effect of the acute toxicity test of white turmeric extract on the histopathological picture of the lungs, no necrosis, congestion, and inflammation was found in the administration of white turmeric extract.