Effect of Sodium Carboxymethylcellulose and Sorbitol on Anti-Peptic Ulcer Activity of Anredera cordifolia Leaves Extract

Anredera cordifolia or binahong is one of the Indonesian medicinal plants that is used to treat peptic ulcer. The purpose of this study was to evaluate the effect of the addition of sodium carboxymethylcellulose (CMC) and sorbitol on anti-peptic ulcer activity of A. cordifolia leaves extracts in male Wistar rats. The plants were extracted using decoction method and freeze dried. Three liquid formulas were used i.e., i) a combination of sodium CMC and sorbitol; ii) only sorbitol; iii) extract only. The rats were divided into 6 groups, i.e., positive control (sucralfate 35 mg/kg body weight); negative control (80% ethanol); normal control; and 3 formulas. After the administration of the liquid formula, all groups, except normal control, were given 80% ethanol (l5 ml/kg body weight) to induce peptic ulcer. Antipeptic ulcer activity was evaluated using direct observation on rats gastric mucosa, and histopathology assessment. The result showed that the strongest anti-peptic ulcer was shown by sorbitol only (96.95% inhibition), followed by the combination of sodium CMC and sorbitol (92.68% inhibition). The formula which only contained extract showed only 31.70% inhibition. Statistical analysis showed significant differences between formula 1 and 2 with negative controls. In conclusion, A. cordifolia leaves extract with the addition of sorbitol showed the strongest anti-peptic ulcer activity. Keyword: Anredera cordifolia, peptic ulcer, suspense, Wistar rat. Introduction Peptic ulcer is a medical condition characterized by discontinuation of the lining or mucosa of gastrointestinal tract due to excessive gastric acid secretion. Approximately 8.41 per 100,000 Indonesians died due to peptic ulcer. The risk factors for peptic ulcers were H pylori infection; chronic use non-steroid anti-inflammatory drugs; excessive alcohol consumption; and tobacco smoking.1-4 A. cordifolia or binahong is one of the Indonesian medicinal plants that are empirically used to treat ulcer. It contains flavonoids, alkaloids, terpenoids, oleanolic acid, saponins, proteins, and ascorbic acid. There is limited research on pharmacological activity of A. cordifolia as an anti-peptic ulcer.5-8 doi: 10.15416/pcpr.v4i1.21392 Corresponding author: Maria Ulfah. Department of Pharmacology, Indonesia School of Pharmacy, Bandung, West Java, Indonesia. Email: maria.ulfah@stfi.ac.id Received: 12 January 2019. Revised: 5 March 2019. Published: 25 April 2019.


Introduction
Peptic ulcer is a medical condition characterized by discontinuation of the lining or mucosa of gastrointestinal tract due to excessive gastric acid secretion.Approximately 8.41 per 100,000 Indonesians died due to peptic ulcer.[3][4] A. cordifolia or binahong is one of the Indonesian medicinal plants that are empirically used to treat ulcer.It contains flavonoids, alkaloids, terpenoids, oleanolic acid, saponins, proteins, and ascorbic acid.There is limited research on pharmacological activity of A. cordifolia as an anti-peptic ulcer.Sodium CMC is one of the most common suspending agent used in liquid formulation.It is hygroscopic, relatively stable in a liquid solution with a pH of 2.0-10.Sorbitol is a preferable wetting agent due to its pleasant sweet taste and can act as stabilizer. 9,10We conducted this study to evaluate the effect of different formulas on anti-peptic ulcer activity of A. cordifolia leaves extract in male Wistar rats.

Materials
The materials used in this study included

Animal model
Animals used in this study were white male Wistar rats aged 2-3 months (150-250 g) Aclimatization was performed for one week before the experiment.

Extraction and formulation of the suspension
A. cordifolia leaves were extracted using decoction method.The ratio between the leaves and water was 1:8.The leaves were boiled for 30 minutes.The extract was then filtered and dried using freeze dryer.The yield was calculated.Phytochemical screening was then conducted to determine secondary metabollites in the extract.

Ulcer in epithelium 3
Three formulas were used i.e., i) a combination of sodium carboxymethylcellulose (CMC) and sorbitol; ii) only sorbitol; iii) extract only.The complete formula of the liquid preparation can be seen in the table 1. Formulas were then evaluated to examine organoleptic properties, pH, viscosity, and specific gravity.

Anti-peptic ulcer activity
The rats were divided into 6 groups, i.e., positive control (sucralfate 35 mg/kg body weight); negative control (80% ethanol); normal control; and 3 formulas.Animal was fasting for 18 hours before the administration of the treatment.One hours after the administration of the treatment, all groups except normal control were given 80% ethanol l5 ml/ kg bw to induce peptic ulcer.After 4 hours, animals were sacrificed. 11Anti-peptic ulcer activity was evaluated using direct observation on rats gastric mucosa, and histopathology assessment.Gastric pH, the number and severity of ulcers, the ratio of ulcer inhibition were measured.Gastric histopathology assessment was performed using hematoxylin staining with eosin.Mucosal damage was measured.Statistical analysis was conducted using analysis of variance (ANOVA).P<0.05 defined statistical significance.0.25 ± 0.500 0.25 ± 0.500 Formula 1 0.5 ± 0.577b 0.5 ± 0.577b Formula 2 0.25 ± 0.500b 0.25 ± 0.500b Formula 3 3.5 ± 2.

Results and Discussion
The extraction of 250 g of A. cordifolia leaves with 2l of distilled water resulted in 46 g of dried extracts (18.4%).Phytochemical screening showed that the extract and all suspensions contained alkaloid, phenol, tannin, quinones, flavonoids, monoterpene sesquiterpene, steroid, and triterpenoide.
Evaluation of characteristics of formula for 1 month is presented in the table 4. The addition of sodium carboxymethylcellulose and sorbitol affected the organoleptic properties, pH, viscosity, and specific gravity.Formula 1 and 2 showed good characteristics.Anti-peptic ulcer activity test was conducted to evalute which formula that showed strongest activity.The strongest ulcer inhibition was shown by formula 2, with 96.95% inhibition.It was followed by formula 1 with 92.68% inhibition, while formula 3 only inhibited 31.70% of the ulcers.
Gastric histopathology observation also showed that formula 2 exhibited better results.In the formula 2, the average gastric mucosal damage was 0.25, indicating no pathological change of the mucosa.Based on Barthel-Spoiled indicator, epithelial desquamation and surface erosion were observed in the ulcer.[13]

Conclusion
A. cordifolia leaves extract with the addition of sorbitol showed the strongest anti-peptic ulcer activity.
380a,c 3.5 ±0.577a,c Description: a = different with normal group, b = different with the positive control, c = different with the negative control Figure 1.Histopathology of Male Wistar Rats Gastric