Violet Leaves Extract To Lower Blood Cholesterol Level Biology Essay

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Cholesterol is a member of a group of substances called steroid which consists a number of essential hormone among the higher animal. Cholesterol also acts as the precursor of the synthesis of many substances.

Having a high cholesterol level proves to be hazardous because extreme arterial pressure can spoil blood vessels in the brain which cause strokes; renal malfunction; or it can cause blindness, deafness, heart attack (in other organs).

The metabolism or chemical reaction of the cellular cholesterol is tightly associated to secure normal cell membrane synthesis with no major intracellular build up. On the other hand, cell may activate to be overloaded cells with lipids such as; Low density lipoprotein (LDL) and cholesterol including cholesterol esters in functioning to the several pathologic processes. Millions of people now a day's are experiencing a higher cholesterol level, excessive intake of cholesterol are one of the reasons. As a group we want to know what should be the other alternative remedy that can use to lower the blood cholesterol level in natural way.

In Atlas of Heart Disease and Stroke report, the World Health Organization liable high cholesterol for causing a third of all cardiovascular diseases worldwide, killing 13 million people each year in developing countries. In the Philippines, 900,000 Filipinos already show signs of stroke, a condition where one out of four victims do not survive the attack, according to a recent National Nutrition and Health Survey.(Malayan, December 3, 2009)

Ferulic acid (FA) is substance originate in the seeds and leaves of most plants, especially in the brans of grasses such as rice and oaths. Ferulic acid has antioxidant component that make it an important anti-aging supplement and it can also give to other potential uses. These consist of application in diabetes, cardiovascular disease, cancer, neuroprotection, bone degeneration, and menopause. Ferulic acid also reduces blood glucose level.

People may have knowledge about cholesterol, like what is the effect of cholesterol when it is in low or high level. Many people can lower their cholesterol level in a cheap way. Violet can be at least an alternative to other drugs that are designed to lower blood cholesterol level.

In dealing with the motivation of this research topic, we are inspired to present our study about the effectivity of the plant Violet (Viola odorata) in lowering blood cholesterol level: this primitive plant may also serve as an anti diuretic, anti pyretic, laxative and sedative.

This research will focus mainly on the effect of Violet (Viola odorata) to lower blood cholesterol level of rabbits (Oryctologous coniculus).

The objective of this study is to establish the potential uses of violet (Viola odorata) like the ability to lower cholesterol level, specifically LDL (Low density lipoprotein).

Statement of the problem:

This study intends to test the effectivity of using Violet (Viola odorata) as an alternative drug to lower blood cholesterol level.

The researchers sought to answer the following questions:

What is the results of blood cholesterol level specifically the LDL prior to the experimental procedure in:

Positive control

Negative control

Experimental group

What is the blood cholesterol level specifically the LDL after the experimentation in the following:

Positive control

Negative control

Experimental group

Is there a significant difference in the cholesterol level of negative control and experimental group?

HYPOTHESIS OF THE STUDY

The following were the hypothesis of the study:

There is no significant difference in the results of blood cholesterol prior to experimental procedure in:

Positive control

Negative control

Experimental group

2. The blood cholesterol levels after the experimentation did not lowered.

There is no significant difference in the cholesterol level between the negative control and the experimental group. Viola odorata is not effective to lower blood cholesterol level.

SIGNIFICANCE OF THE STUDY

Viola odorata contains variable phytochemical components that are promising to study. Many studies have been conducted about various properties of Viola odorata, but little is known about their abilities. However, one of the known properties has a chemical that is the ferulic acid.

Chemical structure of the ferulic acid strappingly resembles that of curcumin which has the ability to lower blood cholesterol level. Curcumin is the substance responsible for the yellow color of the spice turmeric. However, the amount of ferulic acid varies widely depending on the species and growing condition or environment of the said plant. Ferulic acid as well stimulates the elimination of waste products from our body and helps to remove toxins from the blood.

The following person benefits this study:

Medical Technology Student - are now aware of the benefits of Viola odorata.

And have additional understanding on how to conduct a study on the subject of lowering blood cholesterol level.

Community - will be more familiar about Viola odorata. If this study becomes feasible, the use of Violet will then be promoted not only as medicinal for respiratory ailments, but also for blood cholesterol level.

Pharmaceutical - can make this study as one of their basis in a new drug development that will design to lower blood cholesterol level.

SCOPE AND LIMITATIONS:

This study entitled "The Effectivity of using Violet (Viola odorata) to Lower Blood Cholesterol Level" was focus only in cholesterol level which is the LDL. This study aims to find an alternative remedy to lower cholesterol level. The experiment lasts for three (3) weeks, January - February. The experiment such as the preparation of Violet and centrifuged blood samples were done at the College of Medical Technology of Our Lady of Fatima University.

We used ten (10) rabbits (substitute to human) in this research study. In the experiment, we used an ordinary pellet in order for the rabbit to survive. And also a commercial butter (magnolia butter) to increase their blood cholesterol. We utilized Violet root extract to lower their blood cholesterol, specifically the LDL.

Three mL blood samples were extracted from the ear lobes of each rabbits using a butterfly needle. And have it tested at R. De C. Galvez Hospital at Cagayan Valley Road, San Ildefonso, Bulacan.

DEFINITION OF TERMS

LDL. Low-density lipoprotein, 'bad cholesterol'. It accounts for approximately 70% of the circulating cholesterol in an individual.

Control Group. Four rabbits served as control group, two for positive control and two for negative control. Their LDL level served as the reference range for the experimental group.

Experimental Group. Rabbits that are fed with high cholesterol diet and treat with Violet leaves extract.

Negative Control. Rabbits with LDL more than 4.60 mmol/dL. Constantly fed with high cholesterol diet.

Positive Control. Rabbits with LDL that is 1.70 to 4.60 mmol/dL. Oryctologous coniculus fed constantly with normal diet.

Normal diet. Rabbit pellet plus distilled water.

High cholesterol diet. Commercial butter (Magnolia Dairy Cream) that is used to increased blood cholesterol diet.

CHAPTER 2

REVIEW OF RELATED LITERATURE

Related literature

Cholesterol is a soft, waxy, and fat like alcohol found in animal tissues and various foods that is normally synthesized by the liver and is important as a constituent of cell membranes and a precursor to steroids hormones (Soukhanov et.al, 1992).

LDL primarily contains apoB-100 and is more cholesterol rich than other apo-B containing lipoproteins. They form as a consequence of the lipolysis of VLDL. LDL is readily taken up by cells via the LDL receptor in the liver and peripheral cells. In addition, because LDL particles are significantly smaller than VLDL particles and chylomicrons, they can infiltrate into the extracellular space of the vessel wall, where they can be oxidized and taken up by macrophages through various scavenger receptors. Macrophages that take up too much lipid become filled with intracellular lipid drops and turn into foam cells, which are the predominant cell type of fatty streaks, an early precursor of atherosclerosis plaques, (Bishop, 2010).

LDL cholesterol, well validated as a treatable risk factor for CHD (Congestive Heart Disease), is the primary basis for treatment decisions in the NCEP (National Cholesterol Education Program) clinical guidelines. LDL cholesterol = Total cholesterol - HDL -Trig/5. This method, commonly performed as lipid panel, is widely use in estimating LDL cholesterol in the routine clinical practice, having been recommended in the NCEP guidelines, (Bishop, 2005)

Everyday, millions of people inadvertently deposit harmful cholesterol along the arteries that distribute food and oxygen throughout their bodies. Unfortunately, excessive intake of cholesterol may lead to several health problems. The most widely occurring of these is hypertension or commonly known as High Blood Pressure. This condition proves to be dangerous because excessive arterial pressure can destroy blood vessels in the brain to cause strokes; in the kidney to cause renal failure, or in other vital organs to cause blindness, deafness, heart attacks, and so forth (Guyton 1991).

The circulation of blood is a marvel of engineering. It is carried on through a vast network of tiny tubes; most of them are too small to be seen by the unaided eye. To keep the blood in circulation, it must be pumped under a certain amount of pressure (Melloni, 1994).

Certain circulatory disorders trigger the malfunctioning of the heart. One of the major disorders is hypertension, which is the elevation of blood pressure above the normal (Lagua et.al, 1984).

The human body contains 200 - 300 grams of cholesterol while the gallstones may consist entirely of it. Just eating an extra 200mg of cholesterol per 1000 calories a day will take 3.4 years off the life off middle-aged man. That much cholesterol is easy to find in couple of eggs or a pile of bacon (Feltman, 1991).

Viola Odorata (Violet) flowers are said to contain coloring matter, slight trace of volatile oil, three acids, and emetic principles. Dymock and Kirtikar and Basu report that the flower use dry, are especially valued as a diuretic and expectorants, and as a purgative in bilious affectious they consider them antipyretic, and diaphoretic and very useful in relieving febrile symptoms and excitement in all forms of fever, in particularly in combination with other drugs of the same class. Syrup of violet is favorite for cough and hoarseness (Quisumbing, 1978).

Therapeutic and fit for human consumption, the flowers and leaves of violet are prepared into syrup utilized as a substitute remedy for the most part for respiratory ailments connected with congestion, coughing, and sore throat. Flowers are also suitable for eating and used as food additives for instance in salad, made into jelly, and candied for decoration. A decoction made from the root (dry herb) is utilized as a laxative. Tea that is made from the whole plant is utilized to take care of digestive disorders and recent make enquiries has detected the presence of a glycoside of salicylic acid (natural aspirin) which substantiates its use for centuries as a medicinal treatment for nuisance, body pains and as a soothing. Eugenol, Ferulic-acid, Kaempferol, Quercetin, Scopoletin, furthermore demonstrate promise in the management of many kinds of cancer, arthritis, AIDS, gum disease and more. Utilized externally the fresh crushed leaves decrease inflammation and relieve irritations. As a bath additive, the fresh crushed flowers are comforting to the skin and the aroma is very relaxing (http://www.altnature.com/gallery/violet.htm).

Violet Leaf is often made into syrup to relieve respiratory ailments, asthma, colds, coughing, lung congestion and sore throat. Herbalists often prescribe the syrup for Lung and Lymph Cancers

(http://www.mountainroseherbs.com/bu...).

Violet is used as a laxative and diuretic. Being so, it will help to decrease and remove the excess fats in our body. Dietmar also stated that the violet has eugenol, ferulic acid and queritin. In possible cosmetic use of Viola Odorata (Violet) as a bath Additive the fresh crusted flowers are soothing to the skin and the aroma is very relaxing. In essential oil, their flowers also contain volatile oil. It erects emotional imbalance by restoring vegative emotions with positive thought. It is a wonderful formulates oil for relaxing Calming and Meditative needs (Dietmar J. Rummel, 2005).

Ferulic acid has antioxidant properties that make it an important anti-aging supplement, and they also contribute to FA's other potential uses. These include applications in diabetes, cardiovascular disease, cancer, neuroprotection, bone degeneration, menopause, immunity, and (perhaps) athletic performance. Let us look at these uses in more detail (www.naturalnews.com/024084.html).

Ferulic Acid, like many phenols exhibits antioxidant effect in response to free radicals by donating hydrogen from its phenolic hydroxyl group. It has been revealed that the antioxidant capacity of phenolic acid is equivalent to lecithin upon comparison with "Ghee", [a class of clarified butter originated from India] on inhibition of time dependent peroxide value.1) In addition, the reactive oxygen species scavenging effect of ferulic acid has been reported to be similar to that of superoxide dismutase (Gupta S., et al., Milchwissenschaf , 34, 205 (1979). b) Toda S., et al., Plant. Med. , 57, 8 (1991).

Effects of ferulic acid on fertile and asthenozoospermic infertile human sperm motility, viability, lipid peroxidation, and cyclic nucleotides (Free Radic Biol Med. 1997;22(4):581-6. Zheng RL, Zhang H.).

Research flowchart

Conditioning of the Oryctologous coniculus

Preparations of Violet leave extract

Viola odorata

Data Collection

Three experimental rabbits fed a commercial butter

Three experimental rabbits fed a normal diet

Positive control fed a normal diet

Experimental Group

Three rabbits replicates

Negative control fed a commercial butter

Control Group

Two rabbits for Positive & Two rabbits Negative

Ten (10) Rabbits (Oryctologous coniculus) bought at Pethouse Veterinary Clinic at Laguna. Researchers allow the rabbits to recover from the stress for a week. Blood samples were collected to test their cholesterol level, specifically the LDL and serves as the baseline. Violet extract were prepared. Leaves of Viola odorata from Pampanga were washed gently in running water, chopped, get the extract with the aid of a blender, distilled water is used. On the second week of the experimentation, a high cholesterol diet was observed for the rabbits in experimental group. Their blood samples were sent to the laboratory at the end of the week. After force feeding of the Violet extract for the third week, blood samples were again collected and have it tested for the last time to get their LDL level.

CHAPTER 3

RESEARCH METHODOLOGY

RESEARCH DESIGN:

Researchers of this study will investigate the effectivity of Viola odorta to lower blood cholesterol.

The researchers used the experimental research design which is the 'Two-group Design'. The first group is the 'control' which comprises the positive and negative control. Second group is the 'experimental group' which have the replicates. The researchers will strictly follow the procedures necessary to obtain the results of the study.

TEST ORGANISM

Ten (10) male rabbits (Oryctolagus caniculus) of not less four months and not more than eight month old are used in this experiment. And it is bought from Laguna.

Rabbits are place in different cage to prevent food competition.

LOCALE OF THE STUDY

The preparation of Violet extract was conducted at the College of Medical Technology. The extractions of blood samples are done at #38 San Diego St. Marulas, Valenzuela City. The researchers have the samples centrifuge at the College of Medical Technology. The testing of the blood samples were done at the R. De C. Galvez Medical Center (San Ildefonso, Bulacan).

RESEARCH INSTRUMENT AND EQUIPMENT

The materials and samples needed for the experimentation were gathered first to ensure smooth flow of the experimentation. Instruments and equipment used from College of Medical Technology are the pipettes, beaker, centrifuge, blender and weighing balance.

The researchers are conforming to principles, guidelines, and documented sets of procedures in the study to determine the effectivity of Violet odorata to lower blood cholesterol, specifically the LDL.

DATA GATHERING PROCEDURE

Most of the tools and equipment were borrowed from the College of Medical Technology of Our lady of Fatima University.

Ten (10) rabbits (Oryctologous coniculus) are subjected to a series of experiment. The first group will serve as the control group (normal diet of pellets and High cholesterol) while the other will serve as the experimental group (Viola odorata leave extract fed). The researchers allow the Oryctologous coniculus to adjust themselves to their new environment for one week and were given pellets and water for nourishment. All rabbits will have their individual cages to prevent competition for food. The researchers extract 3ml blood (minimum of 2mL) to get the baseline of LDL level. Everytime the Researchers extract blood, butterfly infusion set are being utilized. And extract blood samples from ear lobes.

The control group was given a restricted diet of pellets (average of 50grams/day/rabbit) and water. The experimental group, on the other hand, have been force fed with melted butter (3ml butter/day/rabbit) aside from their restricted diet of pellets. This is to induce high blood cholesterol level. Rabbits are being force fed with butter using a 5ml syringe. This lasts for another week (second week of experiment). Again, the researchers tested for the LDL level for the second time.

For the third week, the researchers force fed the rabbits with the violet extract, with the aid of 5ml syringe. Then, extract again a blood samples to assess their LDL for the last time.

STATISTICAL TREATMENT

ANOVA

One way test

Experimental patient

Sum of Square

Degree

Mean of Square

SD Square

Computed Value

SSC

0.001

1

0.999

1.021

6.9

SSE

1.429

10

0.1429

SST

1.43

11

SSC - Sum of Squares for Column Means

SSC = ∑ ( Txc)2 _ (∑x)2

n N

SSE - Error Sum of Squares

SSE = SST - SSC

SST - Total Sum of Squares

n

SST = ∑ x2 - (∑ x )2

i=1 N

CHAPTER 4

RESULTS AND DISCUSSION

Ten rabbits were bought from Laguna on January 12, 2011. Given with normal diet, researchers allow a three-day acclimation of the rabbits. The first test was done on January 15, 2011 and it served as the baseline. High cholesterol diet was given for a week, and at the end of the week, blood samples were collected to determine their cholesterol level, specifically the LDL. Force feeding of the Violet extract was done at the last week of the experimentation. And LDL level was tested again for the last time.

Positive control

Pre-test

Post-test

Patient 1

4.25

4.15

Patient 2

3.27

3.97

Mean

3.76

4.06

All test results for Pre-test and Post-test of positive control are within the normal range

Negative control

Pre-test

Post-test

Patient 9

6.54

6.55

Patient 10

7.44

7.53

Mean

6.99

7.04

All test results for Pre-test and Post-test of negative control are out of range

Experimental group

Pre-test

Post-test

Patient 3

4.9

4.92

Patient 4

4.72

4.62

Patient 5

4.83

4.8

Patient 6

5.24

5.3

Patient 7

4.97

4.86

Patient 8

5.7

5.71

Mean

5.06

5.04

Pre-test and Post-test for Patients 3, 4, 5, 7, and 8 (experimental group) are out of control while only Patient 6 is in normal range.

Pre-test results of the five rabbits were out of control and only one is in normal range. Post-test of the three rabbits were out of control and the other three are in normal range. Results of the Positive control for both Pre-test and Post-test are in normal range. Both Pre-test and Post-test of the Negative control were both out of control.

All LDL baseline results of the Control group and Experimental group are within the normal range. After a week of feeding the experimental group with high cholesterol diet, the results obtained are above the normal range except for one rabbit. Force feeding of the Violet leave extract was done on the third week (last week) of the experimentation; results obtained have no significant difference from the results after feeding the rabbits with high cholesterol diet.

CHAPTER 5

Summary, Conclusion and Recommendation

Summary of findings

Having a high cholesterol level proves to be hazardous because extreme arterial pressure can spoil blood vessels in the brain which cause strokes and heart attack. The objective of this study is to establish the potential uses of Violet (Viola odorata) like the ability to lower cholesterol level, specifically LDL.

This research entitled "The Effectivity of Using Viola odorata Leaves Extract to Lower Blood Cholesterol Level, Specifically LDL" answered the questions, what is the blood cholesterol level, specifically the LDL after the experimentation in positive control, negative control and experimental group?; is there a significant difference in the cholesterol level of negative control and experimental group?

Base on the test results obtained by the researchers, there is no significant difference in the cholesterol level of negative control and experimental group. From the results obtained, Violet (Viola odorata) leaves extract not proved to be effective in lowering blood cholesterol level of Orytologous conniculus.

This study employed the experimental research method. The experiment utilized Viola odorata that was bought from Arayat, Pampanga. The extract was obtained by the use of a blender where it was liquefied and filtered.

The experiment used 10 male rabbits (Oryctologous conniculus) of the same age and weight which were acquired from Pet House Veterinary Clinic at Laguna. The rabbits were subjected to acclimation for three (3) days and were given a normal diet composed of rabbit pellets and distilled water. After acclimation, they were subjected for blood extraction and tested for total serum cholesterol which served as the baseline. The experimental group was force fed with three mL commercial butter while the control group fed with pellet and water for fourteen (14) days. Then, the total serum cholesterol was tested at R. DeC. Galvez Medical Center at San Juan, San Ildefonso, Bulacan. Before the last test, the experimental group are force fed with Violet leaves extract for seven days and the control group with their normal diet for the last time, blood samples are again collected to test to get their LDL level in the same laboratory.

Unfortunately, Violet leave extract was not an effective to lower blood cholesterol, specifically the LDL level.

Conclusions

In behalf of the findings and results gathered, our research group concludes that the violet leaf extract is not effective for lowering LDL cholesterol. It is due to the reason that the Blood cholesterol levels of the experimental group did not decreased in which there is no significant difference in the negative control and experimental group. Instead, some decreased in a very low value while some are even more increased. However, concluding Violet as ineffective for lowering LDL is the point. Our conclusion is limited to the Violet leave extract only. It is due to the reason that the other parts of the Violet may be effective for lowering LDL. In this case, additional studies to be conducted are recommended.

Recommendations

The researchers would like to recommend further the following:

Additional studies should be conducted on the other parts of the Viola odorata.

Further investigation must be carried out on other phytochemical contents of Viola odorata.

Other tests should be performed regarding Viola odorata results on other cholesterol.

Duration of the experiment should be prolonged.

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