Title:

Chemical Profiling by Thin Layer Chromatography of Sibutramine in Various

Slimming Tea

Introduction

1.1. Background

Lifestyle modification such as proper food intake and exercise maintain the figure of the

body. Conversely, some individual cannot maintain it perhaps due to stress, hectic schedule etc.

In addition, some individual like those who are overweight wants to have a rapid solution in their

problem. Therefore, it is not a surprise that the demand of weight control and slimming product

is increasing. Particularly, slimming tea is one of the herbal supplements that is widely used in

the Philippines in..(add some reasons why). It is readily available over the counter like other

food supplements. Slimming tea is commonly used as laxative. It is used to treat constipation and

also possess properties such as anti-oxidant, anti-inflammatory activities; modulate

detoxification enzymes; stimulate immune function and decrease platelet aggregation. [1]

However, continuous use of slimming tea can lead to laxative dependency will tend to develop

with bouts of chronic diarrhea, abdominal pain, as well as constipation. In the most severe cases,

these laxatives can cause fainting, dehydration and serious electrolyte disorders. [2]

1.2. Sibutramine

In 2011, the FDA of the United States advised the consumers not to purchased or use a certain

slimming tea as it contain an undeclared chemical substance, Sibutramine. [3] ItSibutramine is a

diet aid that works by regulating chemicals in the brain that reduce hunger and craving for food.
[4] Sibutramine is a controlled substance that was removed from the U.S. market for safety

reason in October 2010 [5] The FDA of the Philippines also recalled the products that contain the

said substance between 2010 to 2012. [6]

1.3. Sample Preparation - for what? I guess this should be part of the methdology

Solid phase extraction (SPE) is a technique designed for rapid, selective sample preparation and

purification prior to chromatographic analysis. Using liquid chromatography principles to control

selectivity, SPE provides the sample cleanup, recovery, and concentration necessary for accurate

quantitative analysis. The multitude of available phase chemistries can be packed into an array of

hardware formats such as glass tubes, 96-well plates, and QuEChERS tubes (dispersive SPE),

and can be processed using dedicated vacuum manifold accessories. [7]

1.4. Thin Layer Chromatography

Thin layer chromatography, or TLC, is a method for analyzing mixtures by separating the

compounds in the mixture. TLC can be used to help determine the number of components in a

mixture, the identity of compounds, and the purity of a compound. By observing the appearance

of a product or the disappearance of a reactant, it can also be used to monitor the progress of a

reaction. TLC is a sensitive technique - microgram (0.000001 g) quantities can be analyzed by

TLC- and it takes little time for an analysis (about 5-10 minutes).

The Rf value is used to quantify the movement of the materials along the plate. Rf is equal to the

distance traveled by the substance divided by the distance traveled by the solvent. Its value is

always between zero and one. A TLC analysis might be summarized something like, "Using a

silica gel plate and ethyl acetate as the development solvent, unknown mixture X showed three

spots having Rf's of 0.12, 0.25, and 0.87". Comparing these Rf's with the Rf's of known
compounds might enable a tentative identification to be made. Note that observing three spots

means only that there are at least three components in the mixture. Some components may have

such similar polarities that they appear under one spot after development. [8]

1.5. Significance of the Study interchange this one with your objectives. Yun ang dapat

mauna

As herbal products such as slimming tea is widely used in the Philippines. The relevance of this

study is to ensure the public of their safety regarding the use of slimming tea. In this work, it is

proposed that the method to be used in this study is Thin Layer Chromatography that will detect

the presence Sibutramine in various slimming tea.

* it is suggested that your significance be a bullet like statement. Assess kung sino pa ang

magbebenefit dito like the public, the regulatory agency, etc.

1.6. Objectives of the Study

Generally Objectives: :

To detect if there is a presence of Sibutramine in Slimming tea through the utilization of thin

layer chromatograhyu TLC profiling

Specifical Objectibvesly:

1. To select a proper solvent to be used in extracting slimming tea

2. To separate the chemical substance Sibutramine using Soid Phase Extraction.

3. To compare the standard chemical substance Sibutramine with the various slimming tea

extracted.
1.7. Hypothesis needed pa ba to?

By the end of the study, the researchers seek to find a negative result in the chemical profiling of

Sibutramine in various slimming tea.

1.8. Statement of the problem:

This study aims to test if there is a presence of Sibutramine in various slimming tea to the used

of TLC by analyzing the digitized image data obtained from photographed image of the TLC,

which intends to answer the following questions:

1. Which slimming tea contains the chemical substance Sibutramine.

2. In this study, what parameters can be considered to developed chromatograph presents a good

quality slimming tea extract.

1.9. Scope and Delimitations

This study is limited to chemical profiling of Sibutramine to detect its presence in various

slimming tea particularly available in the Philippine Market. This study will not attempt to

identify the formulation of the various slimming tea.

 Chapter II

Review of Related Literature and Studies

A. Subtitle ? nasan?

Sibutramine (1-[1-(4-chlorophenyl)cyclobutyl]-N,N,3-trimethylbutan-1-amine) usually

as sibutramide hydrochloride monohydrate, is an orally administered agent for the treatment of

obesity. It is a centrally acting stimulant chemically related to amphetamines. Themolecular

formula of Sibutramine is C17H26ClN and molecular weight 279.84804 g/mol. It is a

medicationthatassistswithweightlossbyalteringneurotransmitterswithinthebrain.However,

thecompound was withdrawn voluntarily by the manufacturer from the U.S. market in 2010

because of clinical trial data indicating an increased risk of heart attacks and strokes. Based on

the evaluation of the currently available scientific data, the EMA concluded that the benefits of

sibutramine-containing medicines do not outweigh their risks, and therefore recommended that

the marketing authorisations for sibutramine-containing medicines be suspended across the EU.

Also, the Philippines has already banned the use of Sibutramine in the preparations of slimming

tea due to its toxic effect on cardiovascular.

Parallelly to their withdrawal from the legal market, some of the above mentioned anorexiants

reappeared in illegally adulterated products. Generally, counterfeit anorectic preparations are

claimed to contain only herbal drugs or extracts, however, their efficacy is in fact based on the

presence of marked amounts of synthetic compounds. Beside some functional foods and

traditional Chinese medicines, the majority of the counterfeit products are registered or

unregistered dietary supplements. One of the most frequently applied compounds as adulterant is

sibutramine. In 2009 one article reported that 35 positive samples were found out of a total of
105 botanical dietary supplements, and the most common adulterant was sibutramine in

slimming agents . In some cases sibutramine was detected in Chinese herbal medicines after

observing side effects characteristic of sibutramine.

Sibutraminecanproducedosedependentincreasesinbloodpressureandheartrate,especially

duringinitialtreatment.However,thecardiovasculareffectsofthedrugarerelatedtotheweight

lossachieved:patientswholose5%ormoreofinitialbodyweighthaveareductioninblood

pressure,whichcorrelateswiththedegreeofweightloss.Sibutraminedoesnotexacerbatepre

existingcontrolledhypertensionandtreatmenthasbeenshowntobesafeandeffectiveinthese

patients.Inclinicalpractice,itisimportanttoobservetherecommendedexclusioncriteria,to

monitorbloodpressureandheartrateandtoadheretothewithdrawalcriteria.Thisshouldenable

theidentificationofthosepatientsforwhomsibutramineisnotsuitablewhilepermittingthe

majorityofpatientstogainclinicalbenefitfromtreatmentreatmentSOURCES?

PASTE THE STRUCTURE OF SIBUTRAMINE HERE

2.1. Therapeutic Uses

The U.S. Food and Drug Administration (FDA) is recommending against continued

prescribing and use of Meridia (sibutramine) because this drug may pose unnecessary

cardiovascular risks to patients. FDA has requested that Abbott Laboratories-the manufacturer of

Meridia-voluntarily withdraw this drug product from the United States market. Abbott has

agreed to voluntarily stop marketing of Meridia in the United States. Meridia was FDA-approved

in November 1997 for weight loss and maintenance of weight loss in patients with a body mass

index (BMI) greater than or equal to 30 (=30) kg/sq m or for patients with a BMI =27 kg/sq m

who have other cardiovascular risk factors. BMI is a measure of body fat in adults that is based
on height and weight. Patients with a BMI =30 kg/sq m are considered obese. FDA's

recommendation is based on new data from the Sibutramine Cardiovascular Outcomes (SCOUT)

trial, which demonstrated a 16% increase in risk of major adverse cardiovascular events (a

composite of non-fatal heart attack, non-fatal stroke, resuscitation after cardiac arrest and

cardiovascular death) in patients treated with Meridia compared to patients taking a placebo. At

the end of the trial (60 months), patients in the Meridia group lost a small amount of body weight

compared to patients in the placebo group. FDA has concluded that the risk for an adverse

cardiovascular event from Meridia in the population studied outweighed any benefit from the

modest weight loss observed with the drug. (SOURCES?

2.2. Pharmacologic Action

Sibutramine is an orally administered agent for the treatment of obesity. Sibutramine exerts its

pharmacological actions predominantly via its secondary (M1) and primary (M2) amine

metabolites. The parent compound, sibutramine, is a potent inhibitor of serotonin and

norepinephrine reuptake in vivo, but not in vitro. However, metabolites M1 and M2 inhibit the

reuptake of these neurotransmitters both in vitro and in vivo. In human brain tissue, M1 and M2

also inhibit dopamine reuptake in vitro, but with ~3-fold lower potency than for the reuptake

inhibition of serotonin or norepinephrine. Sibutramine, M1 and M2 exhibit no evidence of

anticholinergic or antihistaminergic actions. In addition, receptor binding profiles show that

sibutramine, M1 and M2 have low affinity for serotonin (5-HT1, 5-HT1A, 5-HT1B, 5-HT2A, 5-

HT2C), norepinephrine (b, b1, b3, a1 and a2), dopamine (D1 and D2), benzodiazepine, and
glutamate (NMDA) receptors. These compounds also lack monoamine oxidase inhibitory

activity in vitro and in vivo. [9]

RESEARCH ON THE PERCEPTION OF THE PUBLIC IN THE USE OF SLIMMING TEA

AND OTHER SLIMMING PREPARARIONS AND DRUGS USE FOR OBESITY.

KONTI NG RRL NYO

RESEARCH ALSO ON THE USE OF TLC IN THIS RESEARCH, SOGNIFICANCE AND

PRACTICALITY OF TLC

Chapter III

Methodology

3.1. Collection of various slimming tea

The sample will be collected in some areas in the Philippines particulary in Luzon. It will be

extracted using an appropriate solvent thru Solid Phase Extraction. After extraction, the sample

will go thru Thin Layer Chromatography.

3.2. Solid Phase Extraction

The purification of sibutramine using Solid Phase Extraction (SPE) allowed the quick separation

of the compound in the fraction eluted from the column with CHCl3. The fraction eluted with

MeOH also contained sibutramine along with herbal constituents.

3.3. Thin Layer Chromatography

The Rf values of the tertiary amine sibutramine in the applied TLC systems were as follows: A:

0.81; B: 0.95; C: 0.93; D: 0.53. Since the most common amphetamine derivatives are primary

and secondary amines, their Rf values of the in TLC systems A, B and C (INCB 2005) are below

0.66, 0.56 and 0.47, respectively, chromatographic analyses in these systems allow the

differentiation of sibutramine. However, due to the high Rf values of sibutramine in these

systems, system D, proposed by us, is more suitable for the analysis of the compound. System D

allowed a good separation of sibutramine from the herbal components of the examined sample,

which were detected below 0.5 Rf values. Among the applied visualization reagents,

concentrated (conc.) H2SO4 was not suitable for the detection of sibutramine. The compound

was not detectable in UV light. Dragendorff reagent resulted in intensive orange spots on yellow

background; with acidified potassium iodoplatinate reagent sibutramine gave dark blue spots.

[10] THIS IS NOT A METHODOLOGY, ILAN ANG SAMPLES NA GAGAMITIN? ANO

ANG SPECS NG TLC? ILAN ANG DEVELOPMENT TIME? SAAN GAGAWIN?

References:

[1] Serafini, M., Rio, D., Yao, D., Bettuzzi, S., & Peluso, I. (2011). Health Benefits of
Tea. CRC Press/Taylor & Francis. Retrieved from
http://www.ncbi.nlm.nih.gov/books/NBK92768/
[2] Wangmo, k., Briceno, R., & COLEsas, G. (2013). The Many Risks of Slimming Tea -
Realnews Magazine. Realnews Magazine. Retrieved 1 July 2016, from
http://realnewsmagazine.net/health/the-many-risks-of-slimming-tea/
[3] Public Notification: Magic Slim Tea Contains Undeclared Drug Ingredient.
(2016). Fda.gov. Retrieved 1 July 2016, from
http://www.fda.gov/Drugs/ResourcesForYou/Consumers/BuyingUsingMedicineSafely/
MedicationHealthFraud/ucm276063.htm
[4] Sibutramine: Indications, Side Effects, Warnings - Drugs.com. (2016). Drugs.com.
Retrieved 1 July 2016, from https://www.drugs.com/cdi/sibutramine.html
[5] Public Notification: Magic Slim Tea Contains Undeclared Drug Ingredient.
(2016). Fda.gov. Retrieved 1 July 2016, from
http://www.fda.gov/Drugs/ResourcesForYou/Consumers/BuyingUsingMedicineSafely/
MedicationHealthFraud/ucm276063.htm
[6] Santos, T. (2016). FDA warns online sellers of banned slimming pills.
Business.inquirer.net. Retrieved 1 July 2016, from
http://business.inquirer.net/159883/fda-warns-online-sellers-of-banned-slimming-
pills
[7] Solid Phase Extraction (SPE) Products for Sample Preparation | sigma-
aldrich.com/spe. (2016). Sigma-Aldrich. Retrieved 2 July 2016, from
http://www.sigmaaldrich.com/analytical-chromatography/sample-
preparation/spe.html
[8] (2016). Retrieved 1 July 2016, from
http://www.chem.umass.edu/~samal/269/tlc.pdf

[9] sibutramine | C17H26ClN - PubChem. (2016). Pubchem.ncbi.nlm.nih.gov.

Retrieved 2 July 2016, from
https://pubchem.ncbi.nlm.nih.gov/compound/sibutramine#section=Literature
[10] (2016). Retrieved 2 July 2016, from
http://www.ingentaconnect.com/content/govi/pharmaz/2013/00000068/00000001/ar
t00003?crawler=true

REVISE YOUR REFERENCES . USE APPROPIRTAE CITATIONS