HEPATOTOXICITY OF ACETAMINOPHEN TREATED BY DELONIX REGIA & AEGLE MARMELOES

44 | P a g e HEPATOTOXICITY OF ACETAMINOPHEN TREATED BY DELONIX REGIA & AEGLE MARMELOES Anil Kumar*, Divya Sharma, Dipayan Purakayastha, Pravesh Sharma, Mohit Khandelwal Department of Pharmacy, Vivekananda Global University, Jaipur 303012, Rajasthan, India. SKIP, Jaipur 303012, Rajasthan, India. Article Info: Received 18 April 2020; Accepted 30 May 2020 DOI: https://doi.org/10.32553/jbpr.v9i3.758 Corresponding author: Anil Kumar Conflict of interest statement: No conflict of interest Abstract Acetaminophen is commonly known as Paracetamol. It is an antipyretic drug, analgesic. It first introduced in 1955 and it cause liver toxicity, which is big issue for Patients now days also. In this work Delnix Regia and Aegle Marmeloes both the plant collected from local area Jaipur and authentify by Department of Botany, Rajasthan University, Jaipur. The authentication number is RUBL 211610 and RUBL 211611and extracted the leaves content by soxhelation process and compare them, which one is more helpful as hepatoprotective because acetaminophen drug cause toxicity in liver and for evaluation purposes different types of test like SGOT, SGPT, Total bilirubine, Total protein, molisch test, wagner test, Hager test, Benedicts test etc performed. In this work we observed that after all evaluation that Delnex Regia is more heptoprotective as compare to Aegle Marmeloes Key word: Acetaminophen, Delnix Regia, Aegle Marmeloes, Hepatotoxicity


Introduction
Acetaminophen (APAP) commonly known as paracetamol (IUPAC name: N-(4-hydroxyphenyl) ethanamide).This medicine is commonly used for antipyretic purpose, pain reliever.Since1955 this drug is available as single dose or with other drug combination, this drug introduced to the market by the McNeil Laboratories in UK. First hepatic failure case due to Acetaminophen comes in UK in 1966.
APAP absorbed rapidly in gastrointestinal tract with peak concentration achieved in 90 minutes, but if food is present in stomach then it may be delay the peak but nut the absorption. APAP undergoes extensive hepatic metabolism. Half life of acetaminophen is around 2-2.5 hour. Acetaminophen is metabolized by glucronidation and sulphate, almost 30% of acetaminophen excreted by urine from body.
Due to lack of anti-inflammatory component, Acetaminophen is out of the NSAIDs family in pharmacological category, it has been always discuss together with these drugs. The discussion on the mechanism of action of Acetaminophen should begin from the analysis of NSAIDs action. Acetaminophen cause liver toxicity in case of overdose of consumption. Every year lots of people died because of liver toxicity due to paracetamol toxicity.
Liver is an important organ which is center of the metabolism of nutrient and excretion of waste material drugs in human body, and it also provide protection body from foreign material by detoxifying them. In case of hepatotoxicity, hepatotoxic chemical damage the liver cell by inducing the lipid peroxidation and oxidative damages. In modern days medicine there is hardly any drug available which can stimulate the liver function or regenerate the liver cells.
Natural medicine like medicinal plants is alternative and safe way for the treatment of the hepatotoxicity. Plan Delonix Regia also commonly known as Gulmohor, family leguminosea sub family fabaceae. This plant is cultivated in topical countries.This plant is useful in antidepressant, anti-bacterial, anti-pyretic analgesic antirheumatic, in this work it used for hepatotoxicity due to acetaminophen poisoning.

Fig-1.2: Liver toxicity
Acetaminophen, a commonly used analgesic and considered as safe therapeutic doses but overdose of Acetaminophen causes severe hepatotoxicity and necrosis in human body. Acetaminophen induced liver toxicity in animals has been used as an experimental model to test the potential hepatoprotective activity by several investigators. The present study is aimed to evaluate the hepatoprotective activity of methanol extract of Delonix regia and Angle Marmelos leaf.

Collection and identification and authentication of plant material:
Leaves of selected plant were collected and authenticated by Department of Botany, Rajasthan University, Jaipur. The authentication number is RUBL 211610 and RUBL 211611. The leaves were shade dried and powdered coarsely in a blender. The powdered plant materials were stored in air-tight container.

Preparation of Ethanolic extracts of Delonix Regia andAegle marmelos leaves leaves :
Plant materials were collected in bulk, washed under running tap water to remove adhering dirt followed by rinsing with distilled water. The plant material was then shade dried and pulverized in a hand mill followed by sieving (sieve no. 40) to obtain coarse powder.
About 190 gm of dry powder were extracted with ethanol (40-60 0 C) for 48 hr in soxhlet extractor. The ethanolic extracts were filtered with Whatman filter paper, concentrated under reduced pressure to a semisolid mass and was made free from solvent. The final obtained extracts were weighed to obtain 28.2g and yield calculated and stored in a cool place.

Benedict's test
To the 150 mg of each extracts, 2ml of Barfoed's reagent was added. Then the mixture was heated on a boiling water bath for 5 min, cooled and observed

Test for alkaloids
To 250 mg of each extracts, 10 ml of dilute HCl was added, mixed and filtered. To the filtrate the following reagents were added and tested.
Wagner's test 2 ml of Wagner's reagent was added to the above filtrate solution and observed.

Hager's test
To the 2 ml of above filtrate solution, 2 ml of picric acid was added and observed

Test for glycosides
The extract was tested for the presence of  Saponin glycosides  cardiac glycosides  Anthraquinone glycosides

Foam test
To 200 mg of each extracts, 15 ml of distilled water added, shake it well and observed.

Legal's test
To 50 mg of each extracts, 1 ml of pyridine, 1 ml of Sodium nitro prusside solution were added and observed.

Keller-kiliani test
To 50 mg of each extracts, 2 ml of glacial acetic acid, 1 ml FeCl 3 solution were added, eated and then cooled. This was transferred to a test tube containing 2ml conc. H 2 SO 4 and observed

Borntrager's test
To 200 mg of each extracts, dil. H 2 SO 4 added and boiled. It was filtered and cooled. To the cold filtrate, 3 ml of benzene added and mixed. The benzene layer separated and to it, ammonia (2 ml) was added and ammonical layer observed.

Lead acetate test
To the 100 mg of each extracts than lead acetate (5 ml) was added and observed.   Table. The level of total protein depleted in the group treated with Acetomenophen (toxin control) and were significantly decreased (P < 0.05) from 6.90g/dl to 5.14 g/dl when compared with the normal control group.

Results
The groups that received the pre-treatment of Delonix regia and Aegle marmelos at dose levels of 400 mg/kg body weight significantly controlled the change in the biochemical parameters. The extracts at dose levels of 400 mg/kg exhibited significant increases (P < 0.05) in the serum total protein level as compared to toxin control group. The total protein and GSH levels from liver homogenate in plant extracts treated groups elevated, but total protein level was not significant. However, pretreatment with Delonix Regia and Aegle marmelos significantly recovered the paracetamol induced Hepatotoxicity in l high dose (400mg/kg) group (P < 0.01, P < 0.05 respectively). The catalase and GPx activity increased of both the extracts; Delonix Regia exhibited good activity (P < 0.01, P < 0.05 respectively). GPx activity of Delonix Regia group was similar to that of standard drug treated group.  Values are Mean ± SEM (n=6) one way ANOVA followed by Dunnett'stest.Where,***p<0.001, **p<0.01 and *p<0.05 and ns-non-significant (a-comparision with control group, )(b-comparision with Acetaminophen control group) Alkaloids Flavanoids Carbohydrates Proteins Saponins - Tannins -+ 7 Anthraquinones Glycosides + + control group, )(b-comparision with Acetaminophen control group).

Conclusion:
Acetaminophen has hepatoxicity issue and in this investigation Delonix Regia and Agles mermelos plant leaves taken and extract through the soxhelation process and by different types of test. In this work we findout that Delnix Regia is more affective as Hepatoprotective compare to Aegle Marmeloes.

Discussion
In the present study; ethanolic extracts of aerial parts of Delonix Regia and Aegle Marmeloes leaves were extracted for the hepatoprotective activity using hepatotoxicity induced by Acetaminophen in rat model and find out the therapeutically better efficacious extract. The shade-dried powder of seeds extracted in a Soxhlet extractor by exhaustive extraction using ethanol as solvent and it gives 17 % yield of extract.
In the present study, leaves of Delonix Regia and Aegle marmelos were evaluate for its pharmacognostical, phytochemical and pharmacological aspects. Acute toxicity testing of both extracts were determine as per OECD guidelines and its ambenment time to time. LD50 of both test extracts were found to be 2000 mg/kg. The degree of hepatotoxicity development can be known by elevated levels of SGOT, SGPT,SALP and Serum Bilirubin enzymes which isattributed due to its covalent bonding N-acetyl-p benzoquinoneimine. Oxidation product of Acetomenophen to sulphydryl group of proteinresulting in cell necrosis and lipid peroxidation induced by decrease in glutathione in the liver,causes hepatotoxicity. Both extracts were Extracted and phytochemical screening done for hepatoprotective activity in albinorats (wistar strains). Hepatotoxicity is clear that when paracetamol wasused to induce liver toxicity there is a substantial increase in enzyme activity of SGOT, SGPT,SALP and Serum Bilirubin. Any decrease in the activity of above enzymes would indicate reversed of induced liver toxicity.