Scientia Naturales Vol.

3 ISSN: 2507-9948 November 2016

Encapsulated Extract of Smallunthus sonchifolius (Yacon)

as Glucose Reducing Agent

Corina Faye B. Ballatong1, Camille Joy B. Palayon1, Queentrexyl V. Peralta1, Frances

Mary B. Ramel1, Ashylyn M. Ventura1 and Jennylin B. Carreon2

1Pharmacy Department, School of Health Sciences

2Centerfor Natural Sciences, School of Arts and Sciences
Saint Mary’s University, Bayombong, Nueva Vizcaya 3700

Abstract. Smallanthus sonchifolius, also known as yacon, is a tea having anti-diabetic

activity which is currently marketed in the Philippines. Several studies have shown
that the leaves of yacon possess different biological effects which include the reduction
of glycemia. The aims of this study were to determine the approximate lethal dose of
the ethanolic extract of yacon leaves based on Approximate Lethal Dose (ALD) by
single-dose method and to investigate the glucose reducing activity of the encapsulated
yacon leaves crude extract (YLCE) on alloxan-induced diabetic mice. Three capsules
were formulated (100, 300 and 500 mg) which contained 25% of the inert ingredients,
lactose and starch, and 75% of the active constituent, the YLCE. The experimental mice
were grouped according to their initial blood glucose level: G1 for negative control
(inert ingredients), G2 for positive control (65 mg Met/kg BW), G3 for 100 mg yacon
capsule, G4 for 300 mg yacon capsule, and G5 for 500 mg yacon capsule. Glucose
reduction assay results showed that the 500 mg yacon capsule had the highest blood
glucose reduction among the three formulations after two hours and after six hours.
Toxicity test showed that the yacon crude extract is safe and non-toxic at 1000 mg/kg
BW dosage. Our results suggest that encapsulated yacon extract can decrease blood
sugar level and the three formulations are safe to use in diabetic mice

Keywords: Smallanthus sonchifolius, encapsulated yacon leaves crude extract,

glucose reducing agent, approximate lethal dose

INTRODUCTION
Diabetes is a chronic metabolic disease reaching an epidemic
proportion in many parts of the world (Lachman, 2003). Many factors like
heredity, age, obesity, diet, sex, sedentary life style, socioeconomic status,
hypertension and various stresses are involved in the etiology of diabetes
mellitus (Khan & Safdar, 2003). Ku and Kegels (2015) stated that the
Philippines is among the 23 low-and-middle-income countries and for the past

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decades, eight of the 10 leading causes of mortality in the Philippines are

chronic conditions and DM type 2 has been consistently among these. The
arising of complications and consequences become alarming that it is likewise
predicted to be among the 10 countries worldwide with the highest numbers
of people with DM type 2 by 2030.
Traditional products are said to be free from side effects, adverse
effects and they are low cost medicines, which will be beneficial for the people
of many countries (Shivanand et al., 2009). Gupta and colleagues (2005)
expressed the interest of using natural products, as a form of supplementary
or alternative medication such as in the case of diabetes. According to Mondal
et al., (2012), some plant-derived drugs lowers blood glucose level and taken
orally are becoming an interest in the treatment of diabetic mellitus that will
prevent complications and adverse effect in many patients. Staines (2011)
stated that holistic approach to health care makes herbal medicine engaging to
people, especially to those seeking herbal remedies to self-treat medical
conditions as with the aim of moving away from the synthetic world towards
a more organic world.
Smallanthus sonchifolius locally named as yacon, is a perennial herb
growing to a height of two to three meters and the flowers are daisy-like with
yellow to orange color. Figure 1 shows the plant parts of yacon.

Fig. 1. Smallanthus sonchifolius

Phytochemical studies have demonstrated that yacon leaves and stems

are rich in proteins and phenolic compounds and flavonoids such as quercetin,
in which one of these secondary metabolites may possess the hypoglycemic
effect (Baroni et al., 2008). Enhydrin is the major sesquiterpene lactone useful
in treating diabetic animals and also effective in reducing post-prandial
glucose (Genta et al., 2010).

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Capsules are solid dosage forms in which medicinal agents and/or

inert substances are enclosed in a small shell of gelatin, it may be hard or soft
gelatin capsules, depending on the composition and they are commonly
employed in clinical drug trials to compare the effects of an investigational
drug with those other drug product in the market and or placebos (Ansel et al.,
2011). Furthermore, in the study of Shivanand et al., (2009), crude extract of
Cedrus deodara was blended with the various excipients for convenience to
formulate as a unit dosage form such as capsule which is one of the bases of
the study.

MATERIALS AND METHODS

Experimental method was utilized in the conduct of the study.
Formulated concentrations of the encapsulated yacon crude extract with inert
ingredients were tested to determine the glucose reduction activity on the
alloxan-induced diabetic mice. The glucose reduction activity of the
formulated yacon crude extract was compared with the effect of the positive
and negative control groups.

Collection of Yacon leaves

The yacon leaves were gathered in Kayapa, Nueva Vizcaya an upland
agricultural farm, situated in the southeastern part of the province of Nueva
Vizcaya. The collected leaves were placed in a clean plastic container and
transported to Saint Mary's University Pharmacy Laboratory. The leaves were
washed thoroughly then placed and stored in a safe place inside the laboratory
for air-drying. The assay of the glucose reduction activity was done in the
university's Center for Natural Sciences (CNS).

Preparation of Yacon Leaves Crude Extracts (YLCE)

Yacon leaves were air-dried and powdered using a laboratory blender.
Five hundred grams (500 g) of yacon leaves were soaked in 90% ethanol for 24
hours. The ethanolic extract was placed in a water bath at 40 - 45°C until the
crude extract was obtained. The crude extract was further formulated into a
capsule dosage form.

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Experimental Animals
Fifteen healthy randomized and properly identified Swiss mice (Mus
musculus domesticus) with a uniform body weight ranging from 18 g to 30 g
were subjected to the experiment. The mice were kept in observation cages in
the CNS Animal House and they were provided with food and water. The
mice were acclimatized for two weeks prior to the conduct of the toxicity
testing and glucose reduction activity. The mice were fasted from food and
water for 24 hours before the test. After the drug administration, the animals
were given free access to food and water. Protocols for the proper handling of
the experimental animals were followed as stated by the Institutional Animal
Care and Use Committee (IACUC) which is promulgated by the Department
of Agriculture Administrative Order No. 40 Series of 1999.

Encapsulation of Yacon Crude Extract

Developing and preparing the formulation
The inert ingredients, 15 mg lactose and 10 mg starch, were mixed and
blended with the use of mortar and pestle. The powder blend was sifted
through a sieve to remove or break up lumps to produce a fine powder.
Seventy-five milligrams (75 mg) of yacon extract was added to the inert
materials and triturated using mortar and pestle.

Filling hard capsule shells

Before filling the capsules, each capsule was tared to counter the
weight of the shell. Using a spatula, the powdered mixture was formed into a
cake having approximately one-fourth the length of the capsule body in a
plate. Then an empty capsule body was held between the thumb and the
forefinger and was punched vertically into the powder cake repeatedly until it
was filled. The capsules were weighed to check uniformity, and then cleaned
individually by polishing them with clean gauze.

Determination of the ALD by Single Dose Method

This Approximate Lethal Dose (ALD) was based on Guevarra (2004).
The maximum volume of the test drug administered to the experimental
animals did not exceed one mL of the test drugs and the control drug
(metformin). The test drugs were dissolved in normal saline solution (0.9%
NaCl) and administered intraperitoneally to the experimental animals at an

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arbitrary initial dose of 10 mg of the test drug per kg body weight (BW) of the
animal, expressed as 10 mg/kg BW. The dose levels were increased
logarithmically at 0.6 log intervals. Then the weight of the formulated
encapsulated YLCE was computed based on the calculated dose levels and the
body weight of the experimental animals. The calculated amount was then
administered until two consecutive doses was obtained in which the lower
dose level should not produce any immediate toxic effects whereas the next
higher dose should be observed to be lethal to the experimental animals.
Observations on the experimental animals were conducted for 24 to 72 hours
post treatment.

Induction of Diabetes in Mice

Alloxan was used to increase the blood glucose level (BGL) of the
mice. 250 mg/kg was dissolved in 1 ml of normal saline solution (0.9% NaCl)
and administered intraperitoneally to the mice which were fasted overnight.
After 24 hours of alloxan induction, blood was collected by cutting the tip of
the tail of the mice and the BGL was determined with the use of a glucometer
(OneTouch®).

Glucose Reduction Activity of the Formulated YLCE Capsule

Glucose reduction activity was determined in the alloxan-induced
diabetic mice by recording their BGL before and after the administration of the
formulated yacon leaves crude extract as encapsulated oral dosage. The
effectiveness of the oral dosage was based on the dosage strength that exerts
the maximum effect in the shortest possible time. The test drugs, positive and
negative controls were dissolved in normal saline solution (0.9% NaCl) prior
to the administration to the experimental mice via oral route. The experimental
mice were grouped according to their initial blood glucose level. Five groups
with five mice each were used in the assay. Group 1 (G1) received the negative
control treatment in which the inert ingredients lactose and starch were used.
Group 2 (G2) received the positive control treatment in which 65 mg Met/kg
BW was administered, Group 3 (G3) was administered with 100 mg yacon
formulation, Group 4 (G4) took 300 mg yacon formulation, and Group 5 (G5)
was orally administered with 500 mg yacon formulation. The alloxan-induced
glucose level of each of the mice was recorded before the assay. Then, the
different treatments were administered and the glucose levels were monitored

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after two hours and six hours. Results of the treatments after 2 h and 6 h of
oral administration were recorded for the percent glucose level reduction
analysis. After the drug administration, blood sample was collected by cutting
the tip of the tail of the test subject and the BGL was determined with the use
of glucometer (OneTouch®). After the assay, the experimental mice were
given free access to food and water.

Oral Glucose Tolerance Test

The procedure in performing oral glucose tolerance test was based on
the study of Aybar et. al.,.(2001) and Genta et. al., (2005). The OGTT was
performed 5 days after treatment for glucose reducing effect of the
encapsulated yacon extract. The formulated 500 mg capsule, positive control
metformin dissolved in one ml normal saline solution and normal saline
solution was administered orally in the 3 group of overnight fast mice. All
mice were loaded with 2g of glucose solution and the blood glucose level was
determined just after the administration and after every 30 minutes for 2
hours.

Percent (%) Glucose Level Reduction

The percent glucose level is computed based on the mean result of the
treatment after two hours and six hours of glucose reduction activity. Percent
glucose was computed using the formula:

% 𝐺𝑙𝑢𝑐𝑜𝑠𝑒 𝐿𝑒𝑣𝑒𝑙 𝑅𝑒𝑑𝑢𝑐𝑡𝑖𝑜𝑛= 𝑖𝑛𝑑𝑢𝑐𝑒𝑑−𝐺 𝑡𝑟𝑒𝑎𝑡ment 𝑥 100

𝐺 𝑖𝑛𝑑𝑢𝑐𝑒𝑑

Where: Ginduced -Alloxan-induced Mice (250mg Allx/kg BW) and Gtreatment -

(65mg met/kg BW, 100 mg tablet, 300 mg tablet, 500 mg tablet).

Animal Disposal
After the different tests, the experimental animals were painlessly
killed by letting them inhale chloroform and they were disposed properly.
This procedure was based according to Guevarra (2005).

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RESULT AND DISCUSSION

Approximate lethal Dose of the Yacon Extract

Approximate lethal dose (ALD) test was used to determine the toxicity
level of the plant extracts. Table 1 presents the results on the toxicity test.

Table 1. Approximate Lethal Dose of Yacon Extract.

Mice BW Dose Calculated mg/kg of extract Observation
(g) mg/kg BW based on BW
1 21.7 10 0.2170 Survived
2 20.4 40 0.816 Survived
3 21.9 160 3.504 Survived
4 26.6 630 16.76 Survived
5 20.7 1000 20.7 Survived
6 (control) 19.8 0 Survived

The data shown in Table 1 revels that the yacon crude extract did not
manifest toxicity even with the highest dose administered which is 1000 mg
crude extract per kg BW of the mice.
Thus, the YLCE is safe and non-toxic. This further indicates that
dosages of 100 mg, 300 mg and 500 mg drug can be safely used in the
formulations of encapsulated oral drugs to reduce BGL. Baroni (2008) claimed
that the oral administration of hydro-ethanolic extract of yacon exhibited low
acute toxicity but no death or adverse effects such as changes in behavior,
posture, exploratory movement, convulsion, abdominal contortions even with
doses up to 5000 mg/kg.

Glucose Reducing Activity of the Yacon Capsules after 2 hours

The three formulated dosages were administered to alloxan-induced
mice with the amount of dosage given to the mice adjusted based on their
body weights. Drug material was administered intraperitoneally and the
blood glucose level of the mice were checked just after two hours.

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Table 2. Glucose Reduction Activity of Formulations after 2 Hours

Dosage formulations N Mean % reduction SD Df F Sig

100mg 3 35.29 8.85

300mg 3 42.36 3.29 2/6 6.402 0.032
500mg 3 54.25 6.32
Total 3 43.97 10.05

Table 2 shows that those mice that were subjected to 500 mg

formulated dosage have the highest mean percentage glucose reduction (x =
54.25, SD = 6.32) compared to mice that were treated with 300mg (x = 42.36, SD
= 3.29) and 100 mg (x = 35.29, SD = 8.85). The apparent difference in the means
was validated by the result of the ANOVA test for significant difference in
mean percent reduction in BGL among the three formulations as the result
yield significant at 0.05 level [F (2/6) = 6.402, p = 0.032]. Accordingly, it can be

Table 3. Multiple Comparisons between the Effect of Metformin and Yacon

Formulations after 2 Hours (post hoc - Gabriel)
Control Drug Formulated Dosages (mg) Mean Diff Sig.
100 21.66 0.18
Metformin 300 14.59 .119
500 2.70 .994

Gabriel Post Hoc multiple comparison test analysis indicates that the
mice treated with metformin have a significantly higher effect than that of the
effect of the 100 mg formulation at .05 level (p = 0.11). However, the effect of
metformin was found comparable to the effects of the 500 mg and 300 mg
formulations.
In the study of Nye and Herrington (2011), metformin is highly
effective in controlling blood sugar level and reducing long-term
complications of type 2 diabetes.

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Glucose Reducing Activity of the Yacon Capsules after 6 hours

The blood glucose levels of the mice were determined for after six
hours of drug administration. Table 4 presents the glucose reduction activity
of the formulations after six hours.

Table 4. Glucose Reduction Activity of the Formulations after Six Hours

Formulated Dosages, mg N Mean SD Df F Sig.
100 3 41.32 1.66 2 9.068 0.015
300 3 48.78 5.64 6
500 3 57.46 5.49
Total 9 49.19 8.07

Table 4 reveals that those mice administered with 500 mg formulated

dosage have the highest mean percentage glucose reduction (x = 57.46, SD =
5.49) compared to mice treated with 300mg (x = 48.78, SD = 5.64) and 100 mg (x
= 41.32, SD = 1.66). The ANOVA test for significant difference in the means
was significant at .05 level [F (2,6) = 9.068, p = 0.015]. Thus it could be inferred
that there is a significant difference in the glucose reduction activity of the
diabetic mice among the 500 mg, 300 mg and 100 mg formulations six hours
after treated with the formulated drugs. This only indicates that the dosage
formulation has something to do with the reduction of blood glucose level.
Table 5 summarizes the result of the ANOVA test for significant
difference in mean percent reduction in BGL between the three yacon
formulations and the metformin 6 hours after they were administered to the
diabetic mice.

Table 5. Glucose Reduction Activity of Metformin and the Three Dosage

Formulations after Six Hours.
Formulated Dosages N MEAN SD D/ F Sig.
Metformin 3 66.35 1.40 3 21.068 .000
100 mg yacon capsule 3 41.32 1.66 8
300 mg yacon capsule 3 48.78 5.64
500 mg yacon capsule 3 57.46 5.49
Total 12 53.48 10.39

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Table 5 indicates that six hours after treatment with metformin,

100mg, 300 mg and the 500 mg dosage, Metformin has the highest mean of
66.35 % followed by 500 mg with a mean of 57.46%. The 100 mg has the lowest
mean percent reduction in BGL (41.32%).
The observed mean difference was found to be statistically significant
based on the analysis of variance result [F (3,8) = 21.068, p = .000]. Accordingly,
it could be inferred that there is a significant difference in the BGL reduction
among the mice when grouped by the treatments.

Table 6. Multiple Comparisons between the Effect of Metformin and Yacon

Formulations after 6 Hours (post hoc - Gabriel)
Control Drug Formulated dosages, mg Mean difference Sig.
100 25.03* .000
Metformin 300 17.57 .004
500 8.89 .136
*The mean difference is significant at the 0.05 level.

Table 6 shows the formulations whose means are significantly

different. Metformin was found to be significantly more effective than that of
the 100 mg and 300 mg formulations six hours after these treatment drugs
were administered. However, the effect of metformin was found comparable
to that of the 500 mg formulation.

Oral Glucose Tolerance Test Result of Yacon

Figure 1 shows the Oral Glucose Tolerance Test (OGTT) of 500 mg
extract. It shows a gradual decrease within the span of two hours and reveals
that encapsulated 500 mg was effective in tolerating glucose within 120
minutes.

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Fig. 2. Oral Glucose Tolerance Test Result of Yacon

Similar result was shown in the study of Miura (2006) wherein mice
treated with yacon extract have shown a significant decrease in blood glucose
level after 60 minutes. This is supported by the study of Park (2009), yacon
tuber extract (YTE) and CGA produced glucose tolerance after one to six
hours.

CONCLUSION
Based on the data and results of the different tests procedures
conducted such as the glucose reduction activity comparing with the three
formulated capsule and metformin and by means of the oral glucose tolerance
test it can therefore be concluded that the encapsulated 500 mg yacon crude
extract has a greater effect than the 300 mg and 100 mg yacon crude extract in
reducing blood glucose to the experimental mice. The effect of the 500 mg
yacon crude extract is similar to the effect of the 65 mg Metformin.

RECOMMENDATION
To further enhance the study, the researchers recommend the
following:
1. To further test the disintegration and stability of the formulated capsule of
yacon crude extract;

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2. To formulate other dosage forms such as tablets and syrups or solutions

with appropriate inert ingredients to the said dosage form that will not
affect the active constituent of yacon; and
3. To evaluate the possibility of the encapsulated yacon leaves crude extract in
clinical studies

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