Tobacco companies discovered that adding ammonia to the cigarette manufacturing process helps convert bound nicotine molecules in tobacco smoke into free nicotine molecules. This process is known as "freebasing." Similar to the chemical process of freebasing cocaine, the end result is an enhanced effect of the drug on the user.
Introduction
Cigarette smoke contains over 4,000 chemicals, including 43 known cancer-causing (carcinogenic) compounds and 400 other toxins. These include nicotine, tar, and carbon monoxide, as well as arsenic, formaldehyde, ammonia, hydrogen cyanide and DDT.
Common additives include yeast, wine, caffeine, beeswax and chocolate. Here are some other ingredients:
Ammonia: Household cleaner
Angelica root extract: Known to cause cancer in animals
Arsenic: Used in rat poisons
Benzene: Used in making dyes, synthetic rubber
Butane: Gas; used in lighter fluid
Carbon monoxide: Poisonous gas
Cadmium: Used in batteries
Cyanide: Deadly poison
DDT: A banned insecticide
Ethyl Furoate: Causes liver damage in animals
Lead: Poisonous in high doses
Formaldehiyde: Used to preserve dead specimens
Ammonia is commonly used for cleaning such as for toilet bowls and windows. But You may be surprised to learn that the tobacco industry has found some additional uses for this household product. By implementing ammonia to your cigarettes, nicotine in its vapor form can be absorbed through your lungs rapidly. This, in turn, result in your brain can get a higher dose of nicotine with each puff.
Nicotine is highly addictive. This is the reason why many people are addicted to smoking. Tar is a mixture of substances that together form a sticky mass in the lungs. Ingredients in tobacco products have never been proved harmful. You may be surprised to learn that the tobacco industry has found some additional uses for this household product. By adding ammonia to your cigarettes, nicotine in its vapor form can be absorbed through your lungs more quickly. This, in turn, means your brain can get a higher dose of nicotine with each puff.
Known as Freebasing Nicotine With Ammonia:
Tobacco companies discovered that adding ammonia to the cigarette manufacturing process helps convert bound nicotine molecules in tobacco smoke into free nicotine molecules. This process is known as "freebasing." Similar to the chemical process of freebasing cocaine, the end result is an enhanced effect of the drug on the user.
Thus by increasing the nicotine kick through the use of ammonia in tobacco processing is said to increase the satisfaction smokers get. Another reason why tobacco companies do freebasing is because it would cause a more intense addictive nicotine experience hence causing a smoker to be easily addicted.
Since we know there’s ammonia inside a cigarette when we smoke. We ought to know what does ammonia does to our body!
Ammonia is a compound of hydrogen and nitrogen, with formula NH3. Under normal circumstances it is a very toxic gas. It is a common precursor for many chemical products, particularly fertilizers and explosives.
Nicotine exist in two forms: Acid(bound) and Base(free). In tobacco smoke, compared to bound nicotine, free nicotine molecules vaporise more easily. Hence when it is vaporised into gas, it could be absorbed quickly and distributed throughout the body.
What are the immediate health effects of ammonia exposure?
Inhalation: Ammonia is irritating and corrosive. When exposed to high concentration of ammonia in air, it causes immediate burning of the nose, throat and respiratory tract. This can cause bronchiolar and alveolar edema, and airway destruction hence leading to respiratory distress or failure. Inhalation of lower concentrations can cause coughing, and nose and throat irritation.
Skin or eye contact: Exposure to low concentrations of ammonia in air or solution may produce rapid skin or eye irritation. On the other hand, higher concentrations of ammonia may result in serious injury and burns. Coming in Contact with concentrated ammonia solutions may cause corrosive injury including skin burns, permanent eye damage or blindness. liquefied ammonia can also cause frostbite injury when contact is made.
Posted on: Tuesday, 31 July 2012 11:52 by Su-Lynn and Adam →
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Ion Exchange Chromatography
Ion Exchange Chromatography
Ion Exchange Chromatography is a common method used to purify the proteins and other charged molecules. In cation exchange chromatography positively charged molecules are attracted to a negatively charged solid support. On the other hand, in anion exchange chromatography, negatively charged molecules are attracted to a positively charged solid support.
Cations are positively charged and Anions are negatively charged.
Cations are positively charged and Anions are negatively charged.
Ion Exchange chromatography - Separation conditions
(A) Adsorption of desired substances
(The substance of interest will bind to the exchanger and unwanted substances are eluted out)
(The substance of interest will bind to the exchanger and unwanted substances are eluted out)
(B) Adsorption of unwanted substances
(The unwanted substances will be adsorbed and substance of interest will pass through the column)
Ion Exchange chromatography -Elution types
(The unwanted substances will be adsorbed and substance of interest will pass through the column)
Ion Exchange chromatography -Elution types
1. Isocratic elution
(Elution with single buffer of constant pH and ionic strength)
(Elution with single buffer of constant pH and ionic strength)
2. Stepwise or gradient elution
(The pH or ionic strength of buffer will change in the process of elution)
(The pH or ionic strength of buffer will change in the process of elution)
Posted on: 10:48 by Su-Lynn and Adam →
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Quantitative Analysis of Ammonia in Whole Tobacco
Quantitative Analysis of Ammonia in Whole Tobacco
Objective:
To isolate and determine the amount of Ammonia in whole tobacco using the High Performance Liquid Chromatography HPLC by external standard method.
Introduction:
Ammonia is one of the many chemicals that exist in a cigarette. One method to separate ammonia is by cation-exchange chromatography using an external standard method. Ion exchange chromatography separates compounds based on net surface charge, which will enable us the identification and quantification of ammonia in cigarettes. Preparation of sample is done by lyophilizing and grinding the whole tobacco. The aim of this experiment is to determine the concentration of ammonia in cigarettes.
Objective:
To isolate and determine the amount of Ammonia in whole tobacco using the High Performance Liquid Chromatography HPLC by external standard method.
Introduction:
Ammonia is one of the many chemicals that exist in a cigarette. One method to separate ammonia is by cation-exchange chromatography using an external standard method. Ion exchange chromatography separates compounds based on net surface charge, which will enable us the identification and quantification of ammonia in cigarettes. Preparation of sample is done by lyophilizing and grinding the whole tobacco. The aim of this experiment is to determine the concentration of ammonia in cigarettes.
Materials and apparatus:
1) Lyophilizer (extract water content from cigarette)
2) Bench top grinder
3) 25,50,100 mL Volumetric flasks
4) High Performance Liquid Chromatograph
5) Solutions needed:
7.1 Ammonium Sulphate > 99 % purity.
7.2 Sulphuric Acid > 96 % purity.
7.3 Methanesulphonic Acid (MSA) – 100 %.
7.4 Type I water (as per ASTM D1193).
Procedure:
(A) Selection of method
In this experiment we will be using HPLC to detect the amount of ammonium ions found in whole tobacco. Concentration of ammonia will then be determined by absorbance measurement.
Column: cation exchange analytical column (50 mm X 4 mm)
Temperature: 30 °C
Mobile Phase / Gradient Elution Conditions (Tertiary Gradient System)
Mobile Phase: 0.003 N (3 mN) sulfonic acid solution
Solvent A: 0.003 N MSA.
Solvent B: Type I water.
Solvent C: 0.2N H2SO4.
Gradient: gradient using 0.2 N (200 mN) H2SO4 to a 0.05 N (50 mN) H2SO4 concentration
Flow: 1.5 mL/minute .
In order to adequately resolve sodium from the ammonium cation for quantitation, a 0.003 N (3 mN) sulfonic acid solution is used as the mobile phase. After the ammonium ion has eluted, a gradient using 0.2 N (200 mN) H2SO4 to a 0.05 N (50mN) H2SO4 concentration is used to remove any divalent cations and quaternary amines that may be present in the sample and may interfere with subsequent samples.
(B) Solution Preparation
For solution preparation, we will prepare the stock standards solution and extraction solution. Preparation of solution A and C will be used in Ion Chromatography.
1) Sulphuric Acid, 0.10N - Stock Standards Solution
1.1) Carefully add approximately 5 g of H2SO4 (96% w/w) to 900 mL of Type I water.
1.2) Mix and dilute to 1 L with Type I water.
2) Sulphuric Acid, 0.025N - Extraction Solution2.1) Carefully add approximately 1g of H2SO4 (96% w/w) to 900 mL of Type I water.
2.2) Mix and dilute to 1 L with Type I water.
3) Sulphuric Acid, 0.20N - Solution C (Ion Chromatography)3.1) Carefully add 10 g of H2SO4 (96% w/w) to 900 mL of Type I water.
3.2) Mix and dilute to 1 L with Type I water.
4) MSA 3mN - Solution A (Ion Chromatography)4.1) Carefully add 0.2 g of Methanesulphonic Acid (MSA) to 900 mL of Type I water.
4.2) Mix and dilute to 1 L with Type I water.
(C) Obtain a representative sample
Tobacco leaves are removed from the selected brand of cigarettes for analysis.
(D) Prepare a lab sample
1) Place approximately 3 g of whole tobacco into a pre-weighed scintillation vial.
Tobacco leaves are removed from the selected brand of cigarettes for analysis.
(D) Prepare a lab sample
1) Place approximately 3 g of whole tobacco into a pre-weighed scintillation vial.
2) Freeze-dry the whole tobacco in a lyophilizer for 48 hrs. Lyophilization (freeze drying) is the removal of water from frozen material.
3) After moisture is removed from the tobacco, grind the whole tobacco in a bench top grinder and weigh accurately 100 mg of ground tobacco into a 16 mL culture tube.
4) Add 10 mL of extraction solution and shake for 60 minutes. Allow mixture to settle for about an hour and filter the solution through a syringe filter into an 8 mL storage vial noting.
5) Using a micropipette, pipette 250 μL of the filtrate into a 2 mL auto-sampler vial and top it up with 1000 μL of extraction solution (1:5 dilution). (This will be our lab sample)
(E) Preparing Standard Solutions
(E) Preparing Standard Solutions
Ammonium stock solution
1) To prepare 25ml for each standard solution, firstly you will need to prepare your ammonium stock solution to be used in the standards. Accurately weigh 0.20 g of ammonium sulphate into a 50 mL volumetric flask
1) To prepare 25ml for each standard solution, firstly you will need to prepare your ammonium stock solution to be used in the standards. Accurately weigh 0.20 g of ammonium sulphate into a 50 mL volumetric flask
2) Dissolve in the Sulphuric Acid 0.10N of H2SO4 that you have prepared earlier on. This will be your ammonium stock solution for the standards.
3) Prepare the standards as followed in the table below:
Standard # | Volume from Ammonium stock solution (μL) | Final Volume (mL) | Concentration [μg/mL] |
1 | 0 | 25 | 0.000 |
2 | 75 | 25 | 3.2693 |
3 | 175 | 25 | 7.6283 |
4 | 250 | 25 | 10.898 |
4) The addition of amount of Ammonium stock solution will be top up to 25ml with your mobile phase (3 mN) sulfonic acid solution. (This will be your standard solution which will be measured.)
(F) Sample Analysis
1) Place the 4 Standard Solutions and the Sample Solution onto the rack in the HPLC auto-sampler. Take note of the location number on your vials on the rack.
2) Program the auto-sampler to inject 5µl portions of your standard solutions and sample solution in separate runs into the HPLC column.
(G) Measuring amount of analyte
1) Determine the peak areas of your standard solutions and sample solution.
2) Plot a Calibration graph of Peak Area (pA*s) against Concentration [μg/mL] and obtain an equation from the graph.
(H) Calculations
1) Calculate the concentration of the sample using the calibration graph to determine the amount of ammonia found in whole cigarette. Remember to multiply back the dilution factor.
2) Program the auto-sampler to inject 5µl portions of your standard solutions and sample solution in separate runs into the HPLC column.
(G) Measuring amount of analyte
1) Determine the peak areas of your standard solutions and sample solution.
2) Plot a Calibration graph of Peak Area (pA*s) against Concentration [μg/mL] and obtain an equation from the graph.
(H) Calculations
1) Calculate the concentration of the sample using the calibration graph to determine the amount of ammonia found in whole cigarette. Remember to multiply back the dilution factor.
Posted on: 09:34 by Su-Lynn and Adam →
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Discussion
In conclusion, the above have shown us how to use HPLC Ion Exchange Chromatography to determine the amount of ammonia in a whole tobacco. The accuracy of the results depends on the correct use of an HPLC column. After the ammonium ion has eluted, perform gradient elution. This is to remove any divalent cations and left over molecules which may interfere with subsequent samples that are injected. Also, the HPLC should be handled with care to extend the life time of a column.
Since we are using the external standard method, the standards and the sample are measured in chromatography separately. The results of the two chromatogram will then be compared. Thus, chromatograohic conditions must be maintained extremely constant. This problem in this experiment is solved by using the auto-sampler. Auto-sampler injection is more accurate compared to the manual injection.
Since we are using the external standard method, the standards and the sample are measured in chromatography separately. The results of the two chromatogram will then be compared. Thus, chromatograohic conditions must be maintained extremely constant. This problem in this experiment is solved by using the auto-sampler. Auto-sampler injection is more accurate compared to the manual injection.
Posted on: 07:21 by Su-Lynn and Adam →
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