Optimization of Senna Alata Leaves Extract for Healing Burns

1. 1.      Introduction

Burns is known as damaged tissue due to contact with agents, thermal, chemical, or electricity. Burns loses its tissue since its interaction with heat sources, such as; water, fire, chemical materials, electricity, and radiation which could inflicts to skin damage and the whole body systems⁽¹⁾. Burns’ healing process could be divided into three phases; (a) inflammation, (b) proliferation, and (c) maturation. The inflammation phase happens since the early occurrence of burn to the seventh days of burn. Then, it continued to the proliferation phase which happens in the end of the inflammation phase until the end of the third week. While the maturation phase happens for several months and could be declared healed after the whole inflammation signs are gone⁽²⁾. Another way in healing burn is by giving plant’s extract to the injured area. Plant is one of the biodiversity which varies and has potential to utilize and develop as a traditional medicine’s basic ingredients⁽³⁾. Senna Alata (Cassia alata L) or known as emperor’s candlesticks is recognized as one of the medical plants used for curing many kinds of diseases. It is warm and has spicy taste. It also contains some chemical compounds, such as tannin, rein aloe-emodina, rein aloe-emodinadiantron, chrysophanic acid, alkaloid, saponin, flavonoid, and anthraquinone glycosides. Basically, the pharmacology effects on Senna Alata (Cassia alata L) could be used as laxative, anthelmintic, anti-itch cream, and drug for skin disorder caused by skin parasite⁽⁴⁾.

1. 2.      Methodology

2.1 Tools and Materials

The tools used in the extraction process were metal with the diameter of 2 x 2 mm, analytical balance, blender, mortar porcelain, filter cloth, spatula, container, razor, brush, digital caliper, maceration tank, rotary evaporator, aluminum foil, rats’ cage, tissue paper, plaster, gauze bandage, hand gloves, and electric oven. Meanwhile, the materials used are ketamine, Senna Alata (Cassia Alata L) leaves extract cream, 96% ethanol, and Vaseline.

2.2 Population and Sample

This study used white rats (Rattus norvegicus) aged 2-3 months, weighing 200-250 gr as the experiment sample. The number of white rats was 30 which then separate in 6 groups. Each groups consists of 5 rats. Here are the groups’ separation detail:

F1        : given 25% concentration of Senna Alata leaves extract ethanol preparation cream

F2        : given 50% concentration of Senna Alata leaves extract ethanol preparation cream

F3        : given 60% concentration of Senna Alata leaves extract ethanol preparation cream

F4        : given 95% concentration of Senna Alata leaves extract ethanol preparation cream

PC       : given burn ointment (Bioplacenton)

NC      : given no treatment

2.3 Phytochemical Screening

Senna Alata (Cassia Alata L) consists of alkaloid, saponin, tannin, flavonoid, and polyphenol compounds.

2.4 The Making of Senna Alata (Cassia alata L) Extract

This study used Maceration methodology for Senna Alata leave’s extraction process. Senna Alata leaves were dried in a room temperature for 1 - 5 days. After it dried, the Senna Alata leaves were cut into small pieces and dried again using oven for an hour with temperature of 50 – 60^o C. The dried leaves then blended using blender until it had smooth powder texture and measured for 1000 gr. The measured leaves then placed into a container and dissolved in 96% ethanol until the whole leaves were sunken. Then covered with cap in order to protect from light and rested for 5 days long with regular stirring for each days. After that, filtered into an empty container and the dregs were macerated back with 96% ethanol. The maceration process was done until 3 times filtering. Thus, the extract was collected and concentrated with rotary evaporator, and the ethanol removed from the extract.

2.5 The Making of Cream and Senna Alata Leaves Extract Preparation

The making of cream using Senna Alata leaves extract was started with the defined measurement which put into mortar and stirred until it homogeny. The extract then, added into each concentrations. Therefore, the preparation cream had concentration of Senna Alata leaves extract for 25%, 50%, 60%, and 95%.

2.6 Open Wound in White rats

a. White rats aged 2-3 months, weighing for about 200-250gr were used for treatment. Before the treatment, the white rats were adapted in laboratory for a week. The rats were given enough food and drink every day in order to keep the weight. The burn injury located on the back of white rats.

b. The first step was shaving the white rats hair located on its back with diameter of 2 cm and disinfected using 70% alcohol. Before shaving the hair, the rats was anesthetized using 0.2 – 0.3 cc doses of ketamine until the rats were unconscious. After that, the hair in the back of the white rats were shaved. Then, the burn was made using metal heated with Bunsen fire for 30 minutes. The hot metal was attached to the rats skin for about 2 – 5 seconds until it formed burn injury, which marked with red color on the skin.        

c. The burns area in the back of white rats were treated with the treatment based on the control groups. The negative control (without giving any treatment), positive control (giving burn ointment or bioplacenton), and the rest four control groups were given the extract of  Senna Alata leaves with the concentration ranged from 25%, 50%, 60%, and 95%.

d. The treatment were done from day one until day twenty two for about two times a day. The wound were left open until it heal. The healing wound was marked with the closed wound.

2.7 Observation on White rats Burns

            This study used quasi experimental design laboratory by observing the white rats three times a day in 21 days. The wound area was measured in the day 3, day 6, day 9, day 12, day 15, day 18, and day 21 from various directions started from day 1 treatment to day 21.

Information:

lx                            : wound area in day x (cm)

lx (1) and lx (2)      : wound area measured from various directions (cm)

1. 3.      Result and Discussion

This is study aimed to analyze the effectiveness of Senna Alata leaves extract toward the healing of burn. The experiment was treated to white rats and conducted in four stages; they were Senna Alata leaves extraction by maceration method, phytochemical test, Senna Alata leaves extract effectiveness test in healing burn in white rats and analyzed the data obtained.

3.1 Extraction results and phytochemical screening

The sample of Senna Alata leaves used was 1200 gr and macerated with 96% ethanol. The weight after immersion was 600 gr and rotated until it became caramel preparation cream weighted for 250 gr. The phytochemical screening of Senna Alata leaves extract and ethanol was illustrated on tables below:

1. Alkaloid identification
2. Steroid and Triterpenoid Identification
3. Saponin Identification
4. Flavonoid Identification
5. Tannin Identification
6. Polyphenol Identification

Based on the tables above, it showed that Senna Alata leaves extract contained with tannin, saponin, flavonoid, alkaloid and polyphenol.

3.2 Optimization Result of Senna Alata Leaves Extract in Healing Burn Based on Treatment Concentration.

            The optimization of Senna Alata leaves extract in healing burn based on treatment concentration (negative control, positive control, 25%, 50%, 60% and 95%) it was conducted by Anova test and the results as follows:

Table 2 above showed that value of sig-p = 0.021 was smaller than 0.05 so that it can be concluded that Senna Alata extract significantly affected wound area. This meant that the application of Senna Alata extraction based on concentration successfully narrowed wound area significantly. In other words, Senna Alata was effective to regenerate burn on white rats’ skin. Moreover, Tukey test was conducted to identify the how much Senna Alata leaves extract concentration that effectively regenerated burn. The results as follow:

Table 3 showed that the subset of group 1 had the largest wound area obtained on K+ treatment for 18.38. Meanwhile, the smallest area for 16.25 was obtained by 95% treatment. It meant that 95% concentration treatment was more optimally healing the burn. However, there was no significant difference between 6 treatments. It was identified by significant value of p= 0.405 was bigger than p= 0.05.

3.3 Optimization of Senna Alata Leaves Extract to Regenerate Based on Treatment Concentration

Based on treatment concentration, ANOVA test result showed value of sig-p = 0.021 was smaller than 0.05. It meant that Senna Alata leaves extract was significantly affected the wound area based on treatment concentration. The most optimal effect was obtained by 95% treatment concentration. It was identified by significant value =0.405 was bigger than 0.05. This experiment was in accordance with experiment entitled Potential Test of Senna Alata Extract (Cassia Alata L.) toward Trichophyton growth inhibition. It was conducted by Noor Hujjatusnaini (2012). The study proven that Senna Alata extract was effectively inhibited Trichophyton sp growth that caused skin disease such as phlegm and ringworm, with a clinical Figure in the form of a skin surface that looks like a sphere - a small circle with red and scaly edges and a slippery center without hair in the area of infection.

The result of this study also in accordance with “Effectiveness test of Ethanol Extract of Senna Alata leaves toward Stomatitis Aphtosa”.  The study showed that flavonoid in Senna Alata leaves could inhibit bacterial growth that caused sprue that identified by the decrease of colony. The higher extract’s concentration, the lower amount of bacterial colony. Some chemical substances in Senna Alata leaves extraxt that could inhibit bacterial or fungus growth were; tannin, rein aloe-emodine, rein aloe-emodina-diantron, chrysophanic acid, alkaloid, saponin, flavonoid and gulcoside antraxion. Generally, pharmacologic effect of Senna Alata leaves was functioned as laxative, worm medicine, itching remover, and skin abnormalities caused by skin parasites

1. 4.      Conclusion and Suggestion

4.1 Conclusion

            Based on the experiment result and analysis, it could be concluded that the extract of Senna Alata leaves (Cassia alata L) gave significance effect toward burns healing in white rats (Rattus norvegicus), either based on the treatment time or the treatment concentration. The most optimal concentration treatment with 95% concentration was succeed in healing or regenerating the burns in white rats with the smallest wound area for 16.25.

4.2 Suggestion

            The writers suggest a similar study done in wider scale by adding the treatment variables and total samples to obtain more accurate results.

Conflict of Interest

Authors declares there is no conflict of interest