The Effect of Oral Administration of Ipomoea batatas L. Leaves Etanolic Extract on Hematological Profile in Rats

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Research J. Pharm. and Tech. 13(2): February 2020

679 ISSN 0974-3618 (Print) www.rjptonline.org

0974-360X (Online)

RESEARCH ARTICLE

**The Effect of Oral Administration of Ipomoea batatas L. Leaves Etanolic Extract on Hematological Profile in Rats**

Anak Agung Bagus Yoga Saputra

^1,2

, Dewa Gede Purnama Putra

^1,2

, I Putu Yogi Astara Putra

^1,2

, Ni Kadek Warditiani

¹

1Department of Pharmacy, Mathematics and Natural Science Faculty, Udayana University, Indonesia.

2Institute Forensic Science and Criminatology, Udayana University, Bali, Indonesia 80361

*Corresponding Author E-mail: [email protected]

ABSTRACT:

Sweet potato (Ipomoea batatas L.) leaves have been shown to have anti-diabetic, anti-inflammatory, anti- mutagenic, antioxidant, anticancer activities, wound healing. However, there is no research on the effect of ethanolic extract of I. batatas L. leaves on hematological profiles; hence, such research needs to be conducted.

The study was carried out by administering a variety of dose of the ethanolic extract of I. batatas L. leaves from maceration results to several groups of rats during 28 days of treatment. Each group consisted of 7 rats with group distribution as follows: normal control (without treatment), negative control (given distilled water), treatment groups (group given extract at a dose of 10 mg/kg body weight, 100 mg/kg body weight, and 1000 mg/kg body weight). The results of an examination of rats' hematological profiles showed that the group given the extract with 1000 mg/kg body weight dose had hemoglobin levels and number of red blood cells that significantly higher (P<0.05) compared to the other groups. Based on this, the ethanolic extract of I. batatas L.

leaves have the potential to increase hemoglobin and erythrocyte levels.

KEYWORDS:

I. batatas L. Ethanolic Extract, Hematological Profile, Rats.

INTRODUCTION:

Indonesia is a country which has a varied diversity of plants. The diversity of these plants is widely used in the health sector to maintain health. One plant that is thought to have a variety of health benefits is I. batatas L.

Phytochemical analysis of I. batatas L. shows that this plant contains anthocyanin, tannins, calcium, magnesium, iron, potassium, manganese, phosphorus, vitamins A, B6, B12, C and D (Johnson and Pace, 2010). Those phytochemical content support I.

DOI:10.5958/0974-360X.2020.00130.4

batatas L pharmacological effects, ranging from anti- mutagenic, anti-diabetic, antibacterial, antifungal, anti- inflammatory, antioxidant, wound healing, anticancer activities. (Panda and Madhav, 2012). Apart from these benefits, there are no studies on the effect of sweet potato leaves, especially those grown in Bali, in affecting hematological profiles. Blood profile — specifically leukocyte count, hemoglobin concentration, number of erythrocytes, and red blood cell indices [including: MCV (Mean Corpuscular Volume), MCH (Mean Corpuscular Hemoglobin), MCHC (Mean Corpuscular Hemoglobin Concentration)]—at normal concentration capable to maintain the human physiological system properly, especially in the immune response and oxygen distribution to all body tissues.

Anthocyanin, as a dark purple pigment of I. batatas L leaves, is a flavonoid compound that has antioxidant activity. The effect of antioxidant on blood has been reported to have a protective effect on blood cells (Fan et al., 2012) and can stabilize sickle red blood cell membranes (Mpiana et al., 2014). Iron content in I.

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batatas L. leaves has been shown to have the ability in enhancing hemoglobin synthesis. Folic acid that also found in I. batatas L. can increase the production of red blood cells (Koury and Ponka, 2004).

According to the beneficial effect of the phytochemical content of I. batatas, L leaves in blood, in this study, an initial screening was carried out to investigate the effect of I. batatas L. leaves on hematological profiles in Wistar strain male rats. The hematological profile observed in this study included: leukocyte count, hemoglobin concentration, number of erythrocytes, MCV, MCH, and MCHC.

MATERIAL AND METHODS:

Experimental Animals and Materials:

I. batatas L. leaves were collected from Aan village, Klungkung regency, Bali. Plant determination was carried out at the UPT Botanical Gardens Conservation Center "Eka Karya" Bali – The Indonesian Institute of Sciences (LIPI). Ethanol 96%, distilled water, citric acid were purchased from PT. Kunia Jaya.

Preparation of I. batatas L. Ethanolic Extract

The collected leaves were sorted to select the dark purple fresh leaves, then washed, steamed for 15 minutes and mashed using a blender. The mashed leaves subsequently extracted with solvent ethanol 77%: citric acid 3% v/v. After two days, the extract was concentrated using a rotary evaporator and continued with the evaporation of the solvent in the oven at 40ºC.

Phytochemical Screening:

The phytochemical constituent of I. batatas L. ethanolic extract was screened to qualitatively detect flavonoid, glycoside, essential oil, alkaloid, polyphenol, tannin, triterpenoid, and saponin. Total anthocyanin content (TAC) was also determined. Iron content in the extract was determined using atomic absorption spectroscopy (AAS).

Treatment of Animals:

The experimental animals used were 35 3-4 weeks of male Wistar rats (approximately 190-200g). The rats were acclimatized for seven days before the experiment.

The rats were divided into five groups, each of which contained 7 test animals. The treatment of each group was as follows: Group A (normal control, no treatment), Group B (negative control, administered orally with distilled water), Group C, D, and E (treated orally with a dose of extract of 10mg/kg body weight, 100mg/kg body weight, and 1000mg/kg body weight, respectively). The rats were freely given access to rat food and water during the 28 days experiment period. All animals were weighed every three days to determine the administered volume of extract.

Hematological Analysis:

Blood collection of rats was carried out on the 29th day.

The rats were anesthetized with ether then 3ml of blood was sampled through the jugular vein using a 3mL syringe. The blood was placed in ethylene diamine tetraacetic acid (EDTA) tubes before the analysis. The hematological analysis was performed using hematoanalyzer to determine the number of erythrocytes, hemoglobin concentration, leukocyte count, MCV, MCH, and MCHC.

Statistical Analysis:

Data obtained were tested for the normality using the Shapiro-Wilk test and its homogeneity using the Levene test. If the data distribution is normal and homogeneous, the analysis is continued to one-way analysis of variance (ANOVA) statistical tests to compare the means of each parameter among the group. The confidence interval was used in all parameter is 95% or P<0.05.

RESULT:

Phytochemical screening shows that the ethanolic extract of I. batatas L. leaves positively contains flavonoids, polyphenols, and tannins. The extract has a TAC of 1562.737mg/L. Meanwhile, the AAS result indicates that the extract contained 3.265ppm of iron.

Figure 1. Profile of Rats’ Hemoglobin (Hb), Leukocyte [White Blood Cell (WBC)], Erythrocyte [Red Blood Cells (RBC)], MCHC (Mean Corpuscular Hemoglobin Concentration), MCH (Mean Corpuscular Hemoglobin), and MCV (Mean Corpuscular Volume)

The rats’ hematological profile after 28 days of an experiment is shown in (Fig.1). The results of ANOVA analysis showed that hemoglobin and red blood cell levels in the group of rats given extract with a concentration of 1000mg/kg body weight were statistically (P<0.05) higher than the other groups. The leukocyte count, MCH, MCHC, MCV, in each group did not show a significant difference (P<0.05).

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681 DISCUSSION:

The results showed that the treatment group administered with the ethanolic extract of I. batatas L.

leaves at a dose of 1000mg/kg body weight had a significantly higher number of red blood cells and hemoglobin (P<0.05) compared to the other groups. This is thought to be caused by the iron content in the ethanolic extract of I. batatas L leaves. Iron is an essential substance in the formation of hemoglobin and erythrocytes. In the formation of erythrocytes, iron is needed in the development stage of erythroblasts (Cavill, 2002). While at the stage of formation of hemoglobin, iron is needed in the formation of heme, where iron reacts with protoporphyrin IX to form heme (Hall and Guyton, 2011).

The higher number of hemoglobin and erythrocytes in the 1000mg/kg body weight dose of treatment was also probably affected by the anthocyanin antioxidant activity contained in the ethanolic extract of I. batatas L.

leaves. According to research by Fibach et al. (2011), antioxidants were able to excite γ-globin gene expression and accumulation of fetal hemoglobin (HbF).

Also, Groom and Kubaisi (2016), reported that anthocyanin is to be able to protect red blood cells from damage caused by free radicals so that the number of hemoglobin is maintained. The amount of hemoglobin in red blood cells decreases by 3% per day due to the process of autoxidation in the red blood cells themselves. The autoxidation process converts hemoglobin to methemoglobin (Murray et al., 2012).

The same was reported in the study conducted by Ologundudu, et al., 2017, where anthocyanin was able to inhibit the decrease in the number of erythrocytes and rabbit hemoglobin due to their protective effects on erythrocytes and hemoglobin against oxidative damage.

Anthocyanin is also known to be able to induce erythropoietin secretion from kidneys, where erythropoietin is the paramount signal for differentiation and proliferation of pluripotent stem cells involved in blood cell formation as reported by Kaur and Kapoor (2005).

The MCV, MCH, and MCHC in Group E did not differ significantly (P<0.05) among the other groups. It indicates that erythrocyte volume, hemoglobin count for each red blood cell, and hemoglobin concentration did not differ significantly (P<0.05) with normal control, so it is considered normal. Ethanolic extract of I. batatas L.

leaves in this study did not affect the number of leukocytes.

The experiment demonstrates that there is a significant difference in the hemoglobin and erythrocyte content at the dose of an extract of 1000mg/kg body weight.

Meanwhile, there are no significant effects of ethanolic

extract of I. batatas L. leaves on leukocyte, MCV, MCH, and MCHC in rats. It can be concluded that effect of ethanolic extract of I. batatas L. leaves on rats’

hematological profile is possibly due to anthocyanin and iron content which have protective activity and enhance the production of erythrocyte and hemoglobin.

ACKNOWLEDGMENT:

The authors acknowledge the authorities of the Department of Pharmacy, Udayana University, for the facilities.

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

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