{"id":60245,"date":"2024-09-30T11:34:33","date_gmt":"2024-09-30T11:34:33","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=60245"},"modified":"2024-10-09T18:00:56","modified_gmt":"2024-10-09T18:00:56","slug":"phytochemical-and-toxicity-evaluation-of-methanol-extracts-of-leaves-and-fruits-of-zanthoxylum-rhetsa-rutaceae","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol17no3\/phytochemical-and-toxicity-evaluation-of-methanol-extracts-of-leaves-and-fruits-of-zanthoxylum-rhetsa-rutaceae\/","title":{"rendered":"Phytochemical and Toxicity Evaluation of Methanol Extracts of Leaves and Fruits of Zanthoxylum rhetsa (Rutaceae)"},"content":{"rendered":"\n

Introduction<\/strong><\/p>\n\n\n\n

Herbal drugs, have been used since time\nimmemorial as food and medicine. Traditional herbal remedies generally use drugs in their natural form\nwith minimal processing and are used to treat diseases that occur locally. In\nlast few years there is a huge surge of interest in these traditionally used\ndrugs because of the belief that they are safe and promote healthier\nliving.  Due to the limitations observed\nwith synthetic drugs, the pharmaceutical industry also has turned to plant\nsources for their drug discovery programmes. Even though there is tremendous\nincrease in use of herbal drugs there is a huge concern about potential\ntoxicities resulting from use of these drugs. Generally, toxicities or adverse\nreactions observed by use of herbs can be due to its inherent toxic nature or\ntheir irrational usage and it\nis very crucial to evaluate the safety of these traditionally used drugs, when\nthey are used in newer forms. Therefore, toxicity studies on plants for\nmedicine or formulations made from them have to be performed before they are\nwidely used1-4<\/sup>.<\/p>\n\n\n\n

Bangladesh, Burma,\nIndia, and the Himalayan region are home to Zanthoxylum\nrhetsa<\/em>, a species of the Rutaceae family, also known as Z. budrunga<\/em> and Z. limonella<\/em>.. It can be found in India’s northeast and along the\neastern and western Ghats5<\/sup>.The fruits are consumed as a spice and the leaves as vegetables in\nIndia’s northern region. Plants have long been used to treat illnesses like\nurinary tract infections and intestinal worms. In addition, it is used as an\nanalgesic and to treat bronchitis, asthma, and cardiac problems.  Fruit-derived volatile oil called mullimam\nhas been utilised as a lipid-lowering, antiseptic, anti-inflammatory, and mosquito-repelling\nagent.  The leaf decoction is used as a\npesticide and to cure intestinal worm diseases by the Naga tribe of\nnortheastern India. Many studies are reported in literature that reveal the\npresence of various classes of secondary\nmetabolites with good biological and pharmacological activities. Zanthoxylum rhetsa<\/em> is widely used in food and therapeutics 6,7<\/sup>.\nHowever, there are no reports in literature\non the systematic evaluation of the toxicity profile of the leaves and fruits\nof Z.rhetsa<\/em>. Thus, the present study\nwas conducted to evaluate acute toxicity studies of the methanol extract of\nleaves and fruits in mice according to Organization for Economic Cooperation\nand Development (OECD) 425 guidelines. <\/em><\/p>\n\n\n\n

Materials and Methods<\/strong><\/p>\n\n\n\n

Collection of plant\nmaterial and preparation of methanol extract<\/strong><\/p>\n\n\n\n

Leaves\nand fruits were collected from Karkala, Udupi district, Karnataka, India.\nDr.\nRajendra D. Shinde of Blatters Hebarium at St. Xaviers College in Mumbai\nvalidated the authenticity of the materials. Fresh fruit and leaves were\nshade-dried, pulverised, A typical maceration procedure was used to prepare the\nmethanol extract. To remove the solvent, the extract was filtered, concentrated\nusing a distillation unit, and evaporated over an electric water bath set at\n40\u00b0C. <\/p>\n\n\n\n

Acute toxicity studies<\/strong><\/p>\n\n\n\n

Total methanol extract of leaf and fruit Z.rhetsa <\/em> was subjected to acute toxicity studies as per\nOrganization for Economic Co-operation and Development guideline for testing of\nchemicals (OECD 420, 2001). To assess the safety of the extracts when taken\norally, a single dose acute oral toxicity trial involving  three rats of the same sex was conducted. <\/p>\n\n\n\n

Animal procurement and acclimatization<\/strong><\/p>\n\n\n\n

The Institutional Animal Ethical Committee of SVKM’s Dr.\nBhanuben Nanavati College of Pharmacy, Vile Parle(W), Mumbai, granted approval\nfor the study. (IAEC approval Number: CPCSEA\/IAEC\/P-78\/2017) The acute toxicity\nwas conducted on 8-12 weeks old female Swiss albino mice weighing 20-25 gm.\nNine animals were procured from National Institute of Biosciences, Pune. The\ncages made of polypropylene held the animals. Three mice per cage in a 12:12\nlight:dark lighting cycle, with controlled temperature (25\u00b12o<\/sup>C) and\nrelative humidity (55\u00b15%). The animals were maintained as per the guidelines of\nthe Committee for the Purpose of Control and Supervision of Experiments on\nAnimals (CPCSEA) India. They were randomly selected and marked on the tail for\nidentification and assigned to three groups. Before beginning the experiment,\nthey had a week to get used to the lab environment.<\/p>\n\n\n\n

Procedure of limit test<\/strong><\/p>\n\n\n\n

Animals were abstained from food overnight prior to dosing but was provided unrestricted access to water. Following the fasting duration, the mice’s body weight was determined and the dosages were calculated. The dosages were freshly prepared and were administered via an oral feeding tube as a suspension using 0.05% of CMC as a solvent at a volume of 1.0 mL\/kg as exemplified below.<\/p>\n\n\n\n

Control group (n=3) administered 0.05% CMC<\/p>\n\n\n\n

Treatment group A (n=3) administered 5000 mg\/kg body weight of methanol extract of foliage<\/p>\n\n\n\n

Treatment group B (n=3) administered 5000 mg\/kg body weight of methanol extract of fruit <\/p>\n\n\n\n

Food was\nprovided after 1 hour of treatment. Throughout the study, all animals were kept\nunder constant observation to assess their health and look for any\nabnormalities. The animals were evaluated for gross behavioural abnormalities,\nclinical symptoms (tremors, convulsions, lethargy, and coma), and mortality at\n30 minutes, 1 hour, 2 hours, and 24 hours. These observations were conducted\nfor 14 days. When two of the three animals showed\nsigns of toxicity or fatality, the dose given proved toxic, and a lower dosage\nwas given to the animals in the following group. To confirm the results, the\nsame dosage was administered again in the event that any one of the three\nanimals showed signs of toxicity or death. The dosage was regarded as safe if\nthere was no fatality. On days 0, 7, and 14, the mean body weights of the\nanimals in the control and test groups were measured. At the end of the\nexperiment, blood was collected by retro orbital route into non heparinized and\nEDTA containing tubes for detailed biochemical and hematological analysis. The\nanimals were killed by cervical dislocation, after which the liver, kidney,\nlung, heart, and spleen were removed and kept for histological analysis in a\nfixation medium containing a 10% solution of buffered formalin.<\/p>\n\n\n\n

Results\nand Discussion<\/strong><\/p>\n\n\n\n

When no other toxicological information is\navailable, acute toxicity testing can offer an initial assessment of a\nmaterial’s toxicity. The median lethal dosage, or LD50, is a useful tool for\nestimating a chemical’s possible risk to humans.  those having an LD50 of less than 5 mg\/kg are\nconsidered very toxic, whilst those having an LD50 of more than 5,000 mg\/kg are\nconsidered basically non-toxic9<\/sup>. <\/p>\n\n\n\n

According to OECD rules, methanol extracts and\nvehicle were administered to the mice in the treatment and control groups of\nthis investigation at a dose of 5000 mg\/kg body weight. For fourteen days\nstraight, they were observed for any indications of toxicity or death.10<\/sup>\n<\/p>\n\n\n\n

Evaluation of mortality, physical and\nbehavioural pattern:<\/strong><\/p>\n\n\n\n

As seen in Table 1.  In the mice, there was no mortality noted.\nThroughout the study period, the mice’s body weight rose (Fig. 1), and their\nskin, fur, and eyes characteristics did not alter (Table 2). These findings\nsuggest that the administration of the crude extract had a small amount of toxicity\non the animals’ growth. \nNo gross changes in behaviour, nor excessive salivation, lethargy,\nsleep, coma or tremors were observed. This showed that treatment with the leaf\nand fruit extracts did not cause any changes in the\ngrowth pattern and metabolism of the mice.11 <\/sup><\/p>\n\n\n\n

Table 1: Toxicity of crude extracts of leaves and fruits in mice.<\/strong><\/p>\n\n\n\n\n\n\n
\n

 <\/p>\n<\/td>\n

\n

Control<\/strong><\/p>\n<\/td>\n

\n

Total methanol extract of leaf<\/strong><\/p>\n<\/td>\n

\n

Total methanol extract of fruit<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Number of dead mice\/ Number of mice used per extract<\/p>\n<\/td>\n

\n

0\/3<\/p>\n<\/td>\n

\n

0\/3<\/p>\n<\/td>\n

\n

       0\/3<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

Table 2: Effect of treatment of methanol extract of leaf and fruits on physical appearance and behavioural pattern in mice<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Observation<\/strong><\/p>\n<\/td>\n

\n

Groups<\/strong><\/p>\n<\/td>\n

\n

30 min<\/strong><\/p>\n<\/td>\n

\n

1 hour<\/strong><\/p>\n<\/td>\n

\n

2 hours<\/strong><\/p>\n<\/td>\n

\n

3 hours<\/strong><\/p>\n<\/td>\n

\n

24 hours<\/strong><\/p>\n<\/td>\n

\n

7 days<\/strong><\/p>\n<\/td>\n

\n

14 days<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Skin and Fur<\/p>\n<\/td>\n

\n

Control<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Leaf<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Fruit<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Eyes<\/p>\n<\/td>\n

\n

Control<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Leaf<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Fruit<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Mucous membrane<\/p>\n<\/td>\n

\n

Control<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Leaf<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Fruit<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Behaviour pattern<\/p>\n<\/td>\n

\n

Control<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Leaf<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Fruit<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Salivation<\/p>\n<\/td>\n

\n

Control<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Leaf<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Fruit<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Lethargy<\/p>\n<\/td>\n

\n

Control<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Leaf<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Fruit<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Sleep<\/p>\n<\/td>\n

\n

Control<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Leaf<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Fruit<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n<\/tr>\n

\n

Diarrhoea<\/p>\n<\/td>\n

\n

Control<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n<\/tr>\n

\n

Leaf<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n<\/tr>\n

\n

Fruit<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n<\/tr>\n

\n

Coma<\/p>\n<\/td>\n

\n

Control<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n<\/tr>\n

\n

Leaf<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n<\/tr>\n

\n

Fruit<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n<\/tr>\n

\n

Tremors<\/p>\n<\/td>\n

\n

Control<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n<\/tr>\n

\n

Leaf<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n<\/tr>\n

\n

Fruit<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n

\n

0<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

1 = Normal and 0= not observed<\/p>\n\n\n\n
\"\"<\/td>\n\n

Figure 1: Effect of methanol extract of leaves and fruits on the body weight of mice<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Evaluation of haematological parameters<\/strong><\/p>\n\n\n\n

Evaluation of haematological parameters is very important to\ncheck the toxicity of plant extract because the blood cells are very\nsusceptible to toxic compounds. Evaluating the impact of the sample on the\nhematopoietic system is also beneficial. Generally, decrease in haemoglobin\ncontent, RBC count and packed cell volume (PCV) is correlated to anaemia and\ndefective haematopoiesis, On the other hand, anaemia is indicated by a rise in\nmean corpuscular volume (MCV) and a fall in mean corpuscular haemoglobin\ncontent (MCHC)12<\/sup>. Results tabulated in Table 3.  revealed \nthat although treatment of both leaves and fruit extract in mice caused\nchanges in the haemoglobin, RBC and PCV content, all values were within CPCSEA\nspecified normal  range for mice 13<\/sup>.\nMice treated with extracts exhibited a decrease in MCV value and increase in\nMCHC content in comparison to control showing that the extract did not cause\nanaemia. WBC content and differential count (count of neutrophils, eosinophil,\nlymphocyte and monocytes ) in extract treated groups and control group were\nwell within the CPCSEA specified normal \nrange normal range thereby exhibiting that the treatment of extracts did\nnot change the haematological parameters of the mice. The content of platelets\nin extract treated groups decreased in comparison to control and it could be\ndue to the thrombolytic nature of the extracts14<\/sup>. Thus, the\nhaematological evaluation showed that the leaves and fruits exhibited minimum\nchanges the parameters indicating the nontoxic nature of extracts.<\/p>\n\n\n\n

Table 3: Effect of Z.rhetsa <\/em>leaf and fruit  <\/em>extract on haematological parameters in acute oral toxicity study.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Parameter<\/strong><\/p>\n<\/td>\n

\n

Units<\/strong><\/p>\n<\/td>\n

\n

Control<\/strong><\/p>\n<\/td>\n

\n

Leaf<\/strong><\/p>\n<\/td>\n

\n

Fruit<\/strong><\/p>\n<\/td>\n

\n

Normal parameters<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Hemoglobin<\/p>\n<\/td>\n

\n

g\/dL<\/p>\n<\/td>\n

\n

9.2 \u00b1 0.14<\/p>\n<\/td>\n

\n

12.2 \u00b1 0.21<\/p>\n<\/td>\n

\n

8.06 \u00b1 0.16<\/p>\n<\/td>\n

\n

10.2 -16.6<\/p>\n<\/td>\n<\/tr>\n

\n

Red Blood Cell<\/p>\n<\/td>\n

\n

million\/ cmm<\/p>\n<\/td>\n

\n

6.16 \u00b1 0.02<\/p>\n<\/td>\n

\n

7.53 \u00b1 0.35<\/p>\n<\/td>\n

\n

5.55 \u00b1 0.25<\/p>\n<\/td>\n

\n

7 -12.5<\/p>\n<\/td>\n<\/tr>\n

\n

White Blood Cell<\/p>\n<\/td>\n

\n

thousands\/ cmm<\/p>\n<\/td>\n

\n

11.7 \u00b10.18<\/p>\n<\/td>\n

\n

9.4 \u00b10.25<\/p>\n<\/td>\n

\n

6.6 \u00b1 0.39<\/p>\n<\/td>\n

\n

6 -15<\/p>\n<\/td>\n<\/tr>\n

\n

Platelets<\/p>\n<\/td>\n

\n

lakh \/cmm<\/p>\n<\/td>\n

\n

945.6 \u00b1 39.71<\/p>\n<\/td>\n

\n

561 \u00b1 46.54<\/p>\n<\/td>\n

\n

649 \u00b1 37.69<\/p>\n<\/td>\n

\n

160 – 410<\/p>\n<\/td>\n<\/tr>\n

\n

Packed Cell Volume<\/p>\n<\/td>\n

\n

%<\/p>\n<\/td>\n

\n

30.46 \u00b1 0.355<\/p>\n<\/td>\n

\n

34.6 \u00b1 0.68<\/p>\n<\/td>\n

\n

33.5 \u00b1 0.35<\/p>\n<\/td>\n

\n

39 – 49<\/p>\n<\/td>\n<\/tr>\n

\n

Mean Corpuscular Volume<\/p>\n<\/td>\n

\n

fL<\/p>\n<\/td>\n

\n

49.7\u00b1 0.64<\/p>\n<\/td>\n

\n

46.33 \u00b1 1.08<\/p>\n<\/td>\n

\n

42.5 \u00b1 0.93<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n<\/tr>\n

\n

Mean Corpuscular Hemoglobin Concentration<\/p>\n<\/td>\n

\n

gm \/ dL<\/p>\n<\/td>\n

\n

30.23 \u00b1 2.18<\/p>\n<\/td>\n

\n

35.2 \u00b1 1.77<\/p>\n<\/td>\n

\n

34.0 \u00b13.08<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n<\/tr>\n

\n

Mean Corpuscular Hemoglobin<\/p>\n<\/td>\n

\n

pg<\/p>\n<\/td>\n

\n

15 \u00b1 0.70<\/p>\n<\/td>\n

\n

16.2 \u00b1 0.34<\/p>\n<\/td>\n

\n

14.4 \u00b1 0.28<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n<\/tr>\n

\n

Neutrophils<\/p>\n<\/td>\n

\n

%<\/p>\n<\/td>\n

\n

27.06 \u00b11.41<\/p>\n<\/td>\n

\n

35.06 \u00b1 2.19<\/p>\n<\/td>\n

\n

29 \u00b1 1.41<\/p>\n<\/td>\n

\n

10-40<\/p>\n<\/td>\n<\/tr>\n

\n

Eosinophils<\/p>\n<\/td>\n

\n

%<\/p>\n<\/td>\n

\n

1 \u00b1 0.0<\/p>\n<\/td>\n

\n

1 \u00b1 0.0<\/p>\n<\/td>\n

\n

0.0<\/p>\n<\/td>\n

\n

0-4<\/p>\n<\/td>\n<\/tr>\n

\n

Lymphocytes<\/p>\n<\/td>\n

\n

%<\/p>\n<\/td>\n

\n

72.15 \u00b1 4.3<\/p>\n<\/td>\n

\n

63.0 1 \u00b1 4.6<\/p>\n<\/td>\n

\n

71.0 \u00b1 3.93<\/p>\n<\/td>\n

\n

55-95<\/p>\n<\/td>\n<\/tr>\n

\n

Monocytes<\/p>\n<\/td>\n

\n

%<\/p>\n<\/td>\n

\n

1 \u00b1 0<\/p>\n<\/td>\n

\n

1 \u00b1 0<\/p>\n<\/td>\n

\n

0.0<\/p>\n<\/td>\n

\n

0.1-3.5<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

Evaluation of serum biochemical parameters<\/strong><\/p>\n\n\n\n

Drug toxicity evaluations involve\nthe assessment of serum biochemical markers, the results of which are summarised\nin Table 4. Serum biochemical indicators of the liver function test include\nalkaline phosphatase, aspartate amino transferase, and alanine amino\ntransferase (ALT, AST). Elevated levels of ALT in serum indicate tissue damage\nand hypertrophy in living organisms. An increase in AST levels is indicative of\nmalfunction in the liver, muscles, and heart. Alkaline phosphatase is a measure\nof the bile duct’s condition. Alkaline phosphatase, ALT, and AST levels are\nhence elevated in liver disorders and hepatotoxicity15<\/sup>. The values\nof  ALT, AST, and alkaline phosphatase in\nthe extract treated groups did not vary to a much in comparison to the control.\nFurther measurement of total protein and albumin showed that it was within  CPCSEA specified range. This exhibited that\nthe methanol extracts of the leaves and fruits did not cause hepatotoxicity. <\/p>\n\n\n\n

Renal function test was  evaluated by measurements of urea and\ncreatinine16<\/sup>. It showed that \nnephrotoxicity was not observed on treatment of extracts. These findings\nwere confirmed by the histopathological observation of the liver and kidney\nwhere no gross morphological changes suggesting lesions were observed.\nMeasurement of serum cholesterol and inorganic ions showed that the extracts\ndid not cause much difference comparison to control.<\/p>\n\n\n\n

Table 4: Effect of Z.rhetsa <\/em>leaf and fruit  <\/em>extract on biochemical parameters in acute oral toxicity study.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Parameter<\/strong><\/p>\n<\/td>\n

\n

Units<\/strong><\/p>\n<\/td>\n

\n

Control<\/strong><\/p>\n<\/td>\n

\n

Leaf<\/strong><\/p>\n<\/td>\n

\n

Fruit<\/strong><\/p>\n<\/td>\n

\n

Normal parameters<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Total Serum Bilirubin<\/p>\n<\/td>\n

\n

mg\/dL<\/p>\n<\/td>\n

\n

0.6  \u00b1 0.16<\/p>\n<\/td>\n

\n

0.4 \u00b1 0.11<\/p>\n<\/td>\n

\n

0.5 \u00b1 0.04<\/p>\n<\/td>\n

\n

0.1-0.9<\/p>\n<\/td>\n<\/tr>\n

\n

SGOT  \/ AST<\/p>\n<\/td>\n

\n

mg\/dL<\/p>\n<\/td>\n

\n

120 \u00b1 6.5<\/p>\n<\/td>\n

\n

131 \u00b17.11<\/p>\n<\/td>\n

\n

128.6 \u00b1 15.5<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n<\/tr>\n

\n

SGPT\/ ALT<\/p>\n<\/td>\n

\n

IU \/ L<\/p>\n<\/td>\n

\n

62 \u00b1 2.44<\/p>\n<\/td>\n

\n

59 \u00b1 4.08<\/p>\n<\/td>\n

\n

74.3 \u00b1 2.0<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n<\/tr>\n

\n

Serum alkaline phosphatase<\/p>\n<\/td>\n

\n

IU \/ L<\/p>\n<\/td>\n

\n

231.3\u00b115.2<\/p>\n<\/td>\n

\n

181 \u00b1 20.7<\/p>\n<\/td>\n

\n

234 \u00b1 11.5<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n<\/tr>\n

\n

Serum total proteins<\/p>\n<\/td>\n

\n

IU \/ L<\/p>\n<\/td>\n

\n

7.1 \u00b1 0.43<\/p>\n<\/td>\n

\n

6.2 \u00b1 0.14<\/p>\n<\/td>\n

\n

6.6 \u00b1 0.29<\/p>\n<\/td>\n

\n

3.5-7.2<\/p>\n<\/td>\n<\/tr>\n

\n

Serum total albumin<\/p>\n<\/td>\n

\n

g \/ dL<\/p>\n<\/td>\n

\n

3.4 \u00b1 0.28<\/p>\n<\/td>\n

\n

4.2 \u00b1 0.09<\/p>\n<\/td>\n

\n

3.9 \u00b1 0.57<\/p>\n<\/td>\n

\n

2.5-4.8<\/p>\n<\/td>\n<\/tr>\n

\n

Blood urea nitrogen<\/p>\n<\/td>\n

\n

g \/ dL<\/p>\n<\/td>\n

\n

14.1 \u00b1 0.66<\/p>\n<\/td>\n

\n

18.3 \u00b1 1.2<\/p>\n<\/td>\n

\n

12.46 \u00b1 1.3<\/p>\n<\/td>\n

\n

12-28<\/p>\n<\/td>\n<\/tr>\n

\n

Serum creatinine<\/p>\n<\/td>\n

\n

mg \/ dL<\/p>\n<\/td>\n

\n

0.4 \u00b1 0.14<\/p>\n<\/td>\n

\n

0.7 \u00b1 0.08<\/p>\n<\/td>\n

\n

0.4 \u00b1 0.21<\/p>\n<\/td>\n

\n

0.3-1<\/p>\n<\/td>\n<\/tr>\n

\n

Random blood sugar<\/p>\n<\/td>\n

\n

mg \/ dL<\/p>\n<\/td>\n

\n

98.33 \u00b12.4<\/p>\n<\/td>\n

\n

107 \u00b1 2.16<\/p>\n<\/td>\n

\n

88 \u00b10.81<\/p>\n<\/td>\n

\n

62-175<\/p>\n<\/td>\n<\/tr>\n

\n

Serum cholesterol<\/p>\n<\/td>\n

\n

mg \/dL<\/p>\n<\/td>\n

\n

78.33 \u00b12.05<\/p>\n<\/td>\n

\n

72.0 \u00b12.94<\/p>\n<\/td>\n

\n

75 \u00b1 4.08<\/p>\n<\/td>\n

\n

26-82<\/p>\n<\/td>\n<\/tr>\n

\n

Serum calcium<\/p>\n<\/td>\n

\n

mg\/dL<\/p>\n<\/td>\n

\n

7.9 \u00b1 0.29<\/p>\n<\/td>\n

\n

8.2 \u00b1 0.24<\/p>\n<\/td>\n

\n

8.1 \u00b1 0.28<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n<\/tr>\n

\n

Serum phosphorous<\/p>\n<\/td>\n

\n

mg\/dL<\/p>\n<\/td>\n

\n

4.2 \u00b1 0.32<\/p>\n<\/td>\n

\n

3.9 \u00b1 0.69<\/p>\n<\/td>\n

\n

4.8 \u00b1 0.77<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n<\/tr>\n

\n

Serum sodium ion<\/p>\n<\/td>\n

\n

mEq\/L<\/p>\n<\/td>\n

\n

140.3 \u00b1 3.68<\/p>\n<\/td>\n

\n

138.76\u00b11.8<\/p>\n<\/td>\n

\n

140.5 \u00b1 0.41<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n<\/tr>\n

\n

Serum potassium ion<\/p>\n<\/td>\n

\n

mEq\/L<\/p>\n<\/td>\n

\n

3.4 \u00b10.21<\/p>\n<\/td>\n

\n

4 \u00b1 0.71<\/p>\n<\/td>\n

\n

4.13 \u00b1 0.24<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n<\/tr>\n

\n

Serum chloride ion<\/p>\n<\/td>\n

\n

mEq\/L<\/p>\n<\/td>\n

\n

102 \u00b1 2.9<\/p>\n<\/td>\n

\n

104.8\u00b13.02<\/p>\n<\/td>\n

\n

103.7 \u00b1 4.00<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

Histological evaluation<\/strong><\/p>\n\n\n\n

Histological evaluation helps in\nidentification of   toxicity of compound\nat the organ or tissue level. As observed under light microscope (Fig.2)\ntreatment with extracts did not cause any abnormalities or lesions in the\nmorphology of the tissues of the major organs such as heart, kidney, lung,\nliver and spleen. No hypertrophy was found in the tested organs. Hypertrophy is\nan indication of toxicity in the organs 17<\/sup>. <\/p>\n\n\n\n

During the 14-day study, the animals did not appear any\nmortality or change in body weight. There were no changes in the behavioural or\nmotor neural functions of the body.  The\nhaematological and serum biochemical parameters along with the\nhistopathological studies also did not exhibit any abnormalities or\npathological changes in the major organs. Consequently, it was discovered that\nthe leaf and fruit extract’s LD50 (oral fatal dose) was greater than 5000 mg\/kg\nbody weight.<\/p>\n\n\n\n

The findings of acute toxicity tests executed on Z. rhetsa<\/em> leaves and fruits matched\nthose of toxicity tests done on other Zanthoxylum species. In the past, it has\nbeen found that Z.armatum<\/em> fruit and\nleaf extracts were safe at doses up to 2000 mg\/kg body weight 18<\/sup>.\nSimilarly, it has been found that a methanol extract of F. zanthoxyloides<\/em> root bark was nontoxic up to 5.0 g\/kg of rats’\nbody weight19<\/sup>. <\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 2:<\/strong> Histopathology of the lung, spleen, heart, liver and kidney of mice in acute toxicity test at 40x magnification (haematoxylin-eosin stain)<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Conclusion<\/strong><\/p>\n\n\n\n

Zanthoxylum\nrhetsa<\/em> leaves and fruits are\nwidely used in food and therapeutics. Therefore, toxicity tests must be carried\nout in order to evaluate the safety profile of Z. rhetsa<\/em> leaves and fruits. In compliance with OECD necessities,\nresearch on the acute toxicity of methanol extract of leaves and fruits has\nbeen carried out. From the\nresults obtained, it can be concluded that Z.rhetsa<\/em>\ndid not exhibit any toxic effects even when used at a very high\nconcentration.  Thus, the leaves and\nfruits can be safely explored for its use in therapeutics and drug discovery\nresearch. Further genotoxicty and mutagenicity  \nstudies can be conducted in vivo<\/em>\nto establish complete safety profile of the drug.<\/p>\n\n\n\n

Acknowledgements<\/strong><\/p>\n\n\n\n

None<\/p>\n\n\n\n

Conflict of Interest<\/strong><\/p>\n\n\n\n

There is no conflict of interest<\/p>\n\n\n\n

Funding Source<\/strong><\/p>\n\n\n\n

There are no funding sources<\/p>\n\n\n\n

References<\/strong><\/p>\n\n\n\n

  1. Tilburt J.C., Kaptchuk T.J. Herbal medicine research and global health: an ethical analysis. Bull World Health Organ. 2008; 86:594\u2013599.
    CrossRef<\/a><\/li>
  2. Yuan H., Ma Q., Ye L., Piao G. The Traditional Medicine and Modern Medicine from Natural Products. Molecules. 2016; 21:559. doi:10.3390\/molecules21050559
    CrossRef <\/a><\/li>
  3. Ekor M. The growing use of herbal medicines: issues relating to adverse reactions and challenges in monitoring safety. Frontiers in  Pharmacology. 2014; 4:177.
    CrossRef <\/a><\/li>
  4. Ping K.Y., Darah I., Chen Y., Sreeramanan S., and Sasidharan S. Acute and subchronic toxicity study of Euphorbia hirta<\/em> L. methanol extract in rats, BioMed Research International. 2013; 2:182064\u00b7
    CrossRef <\/a><\/li>
  5. Mallya R. and Bhitre M.J. Pharmacognostic standardisation and chromatographic fingerprinting of leaves and fruits of Zanthoxylum rhetsa<\/em>. International journal of pharmacy and pharmaceutical sciences. 2018; 10:101-104.
    CrossRef <\/a><\/li>
  6. Anonymous. The Wealth of India: Raw Materials and Industrial Products. Publication and Information Directorate, CSIR, New Delhi, Vol. XI,  1956, pp.22-23.<\/li>
  7. Quattrocchi U. CRC world dictionary of medicinal and poisonous plants: common names, scientific names, eponyms, synonyms, and etymology, CRC Press, India;2012, pp.3650.<\/li>
  8. OECD. OECD Guideline for Testing of Chemicals: Acute Oral Toxicity-Fixed Dose 17 Procedure (No.420). OECD, Paris;2001<\/li>
  9. Erhirhie E.O, Ihekwereme C.P., Ilodigwe E.E. Advances in acute toxicity testing: strengths, weaknesses and regulatory acceptance. Interdisciplinary toxicology. <\/em>2018;11:5\u201312. doi:10.2478\/intox-2018-0001
     CrossRef <\/a><\/li>
  10. Jothy S.L., Zakaria Z., Chen Y., Lau Y.L., Latha L.Y., Sasidharan S. Acute Oral Toxicity of Methanolic Seed Extract of Cassia fistula<\/em> in Mice. Molecules. 2011; 16:5268.
    CrossRef <\/a><\/li>
  11. Sreeramanan S., Sasidharan S.. Acute and subchronic toxicity study of Euphorbia hirta<\/em> L. methanol extract in rats. Biomedical Research International. 2013; 2013:182064.
    CrossRef <\/a><\/li>
  12. Chanda S., Parekh J., Vaghasiya Y., Dave R., Baravalia Y. and Nai R: Medicinal Plants – From Traditional Use to Toxicity Assessment. A Review  International  Journal of  Pharmacetical Sciences and Research. 2015; 6:2652-2670. <\/li>
  13. CPCSEA. Compendium of CPCSEA. New Delhi. 2018. http:\/\/thsti.res.in\/ pdf\/Compendium_CPCSEA_2018-I.pdf. Accessed June 5, 2019.<\/li>
  14. Afolayan A.J., Wintola O.A., Fouche G. Acute and Subacute toxicological evaluation of the aerial extract of Monsonia angustifolia<\/em> E. Mey. ex. A. rich in Wistar Rats. Evidence Based Complementary and Alternative Medicine. 2016; 2016:4952485.
    CrossRef <\/a><\/li>
  15. El Kabbaoui M., Chda A., El-Akhal J. Acute and sub-chronic toxicity studies of the aqueous extract from leaves of Cistus ladaniferus<\/em> L. in mice and rats. Journal of Ethnopharmacology. 2017; 209:147-156.
     CrossRef <\/a><\/li>
  16. Kuatsienu L.E., Ansah C., Adinortey M.B. Toxicological evaluation and protective effect of ethanolic leaf extract of Launaea taraxacifolia <\/em>on gentamicin induced rat kidney injury. Asian Pacific Journal of Tropical Biomedicine. 2017; 7: 640-646.
     CrossRef <\/a><\/li>
  17. Mawoza T., Tagwireyi D. and Nhachi  C: Acute and sub-chronic toxicity studies of an aqueous stem bark extract of Sclerocarya birrea<\/em> using a rat model. International Journal of Pharmaceutical Sciences and Research. 2016; 7:9-17. <\/li>
  18. Barkatullah, Muhammad I., Muhammad  N. Evaluation of Zanthoxylum armatum<\/em> DC for in-vitro<\/em> and in-vivo<\/em> pharmacological screening. African Journal of  Pharmacy and Pharmacology. 2011; 5:1718-1723.
     CrossRef <\/a><\/li>
  19. Ogwal-Okeng J.W., Obua C., Anokbonggo W.W: Acute toxicity effects of the methanolic extract of Fagara zanthoxyloides<\/em> (Lam.) root-bark. African Health Science. 2003; 3:124-126.    <\/li><\/ol>\n","protected":false},"excerpt":{"rendered":"

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