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Module 4 Alcohol and the Breathalyzer Test MODULE 4 - OVERVIEW Description of the module Once alcohol reaches the bloodstream, it equilibrates with different body tissues, including the lungs. Alcohol is a volatile molecule that can be eliminated from the body in exhaled air. Students will learn how alcohol equilibrates with lung air spaces, and how to estimate how much alcohol is in the blood based on what is in the exhaled air. The module covers concepts in biology, chemistry, and math including: 1) passive diffusion across a biological membrane 2) the basic anatomy of the circulatory system 3) the chemical principles of volatility 4) dynamic equilibrium of a biological system 5) oxidation and reduction reactions 6) the use of ratios in scientific calculations Content Standards CA1, CA22, CA23, CA25, CB3, CB24, CB30, CB32, CB34, CE1, CE2, CE21, CF13, CF63, CG21, AA1, AA2, AA3, AA4, AB1, AB11, AB12, AB13, AB14, AB15, AC2, AC4, AC5, AD2, AD4, TA2, TA3, TA4, TB1, TB2, TB3, TB5, TC1, TC2, TC3, TC4, TC5, TD2, TE4, TE6, TF3 MODULE 4 - LEARNING OBJECTIVES Learning objectives After participating in this module, students will be able to: Illustrate the path that a molecule would take to enter the circulation from the stomach/intestine and be transported throughout the body. Recognize that soluble compounds are dissolved in the blood. Explain the relationship between passive diffusion and the concentration of a molecule. Distinguish between active transport and passive diffusion. Identify the chemical and physical properties of an alcohol that determines its volatility. Discuss the term "dynamic equilibrium" as it applies to a biological system, using ethanol as an example. Recognize the oxidized and reduced forms of a molecule. Relate chemical reactions to real-life applications (e.g., use the breathalyzer test as an example). Calculate the blood alcohol concentration (BAC) using a blood-to-breath ratio. After participating in this module, it is expected that students may be able to: Extend this knowledge to predict the path of a drug to its target organ in the circulatory system depending on its route of administration. Compare and contrast the volatility of compounds other than ethanol. Evaluate the accuracy of the breathalyzer test for intoxication using scientific principles and mathematical formulas. Explain the scientific principles of the breathalyzer test or other commonly used breath tests for alcohol intoxication. MODULE 4 - STUDENT HANDOUT Spritney Beers Blows it Bigtime The big news is that celebrity singer, Spritney Beers has been arrested for driving under the influence (DUI). Spritney blew a "0.08%" on the Breathalyzer™ when she was stopped by the police after driving over the center line. This value reflects the blood alcohol concentration (BAC) and, in California, it is the value that legally designates intoxication. Despite considerable evidence that the Breathalyzer™ test gives an accurate approximation of the BAC, there is still much discussion about the validity of this test. While in jail, Spritney used her one phone call to contact her attorney. A court date has been set for three weeks from today. Paparazzi photographed her as she left the courthouse in a heated discussion with her attorney. Reporters at the scene quoted him saying "I am very suspect of the validity of the Breathalyzer™ test given to my client. I only have to establish reasonable doubt in this case." Back at school, you are discussing the case with a classmate when your science teacher interrupts and decides that the topic is worthy of a class debate. To prepare for the class debate, you decide to learn how a Breathalyzer™ test is used to determine the BAC so that you can debate whether it is accurate or not. Spritney Beers' level of alcohol intoxication was assessed by measuring the concentration of ethanol she expelled from her lungs. This technique works because a water-based molecule like ethanol that is absorbed from the gut into the bloodstream will reach the lungs, where it is exhaled as a vapor in the air. 1. Diagram the path that alcohol will take from the gut (stomach & small intestine) to the lungs via the circulatory system. Which membranes will ethanol have to cross? Although some alcohol is exhaled from the lungs, the rest stays in the blood. Where does the ethanol in the blood go once it leaves the lungs? Ethanol is dissolved in the blood and is distributed to organs around the body. It is a volatile molecule and can be vaporized quite easily. In the lung, ethanol is converted from a liquid to a gas, so it can be exhaled in the air. 2. Identify and describe the chemical and physical properties of ethanol that contribute to its volatility. It will help to draw the chemical structure of ethanol. 3. Where in the lung is ethanol vaporized? What role does the lung play in the vaporization of ethanol? The Breathalyzer™ can approximate a person's blood alcohol concentration (BAC) because the concentration of alcohol vapor in the lungs is directly related to its concentration in the blood. Alcohol vaporizes in the air sacs (alveoli) of the lungs and achieves an equilibrium with the concentration of alcohol that is still in the blood. 4. What is meant by equilibrium? Are ethanol molecules still moving across the membrane between the capillary and the alveolus? 5. What would happen to the equilibrium if some of the ethanol leaves the alveolar sac by exhalation? When a person exhales into a breath analyzer such as the Breathalyzer™ tube, the exhaled alcohol reacts with compounds in the Breathalyzer™ chamber to produce a change in color from orange to green. The chemical reaction indicated by the color change involves oxidation and reduction. Silver nitrate catalyzes the reaction 6. What is oxidized in the Breathalyzer™? What is reduced? What role does the catalyst play in the chemical reaction? The degree of the color change indicates how much alcohol is present in the expired air and the instrument calculates an actual concentration. However, for the Breathalyzer™ to calculate how much alcohol is present in the expired air sample it must take into account the volume of blood from which the alcohol originated. There is a standard way of describing this relationship; it is called the blood-to-breath ratio or partition ratio. The average blood-to-breath ratio is 2100:1 and this is the value used for legal purposes. The ratio assumes that an equilibrium exists between the blood and the alveolar air. 7. Explain what this ratio means in terms of the concentration of alcohol in the blood and the breath. Does the ratio change as one exhales? Actually, the ratio can vary between 1500:1 and 3000:1 depending upon a number of factors including a person's age, gender, and genetic makeup. In Spritny Beers' case, the reported BAC was 0.08%, based on the 2100:1 ratio. In fact, she may be anywhere in this range from 1500:1 to 3000:1. 8. Calculate the underestimation and overestimation of Spritney's BAC assuming she had a bloodto-breath ratio of 1500:1 and 3000:1. Would Spritney still be considered legally intoxicated based on your answers? As a point of discussion, consider the advantages and disadvantages of using the Breathalyzer™ test and decide for yourself whether you would prosecute based on the Breathalyzer™ evidence in this case. Is there other evidence to consider? MODULE 4 - CONTENT A - How does ethanol get to the lungs? B - Ethanol vaporizes to a gas in the lungs C - Ethanol leaves the lungs in the air D - Ethanol in the blood equilibrates with ethanol in the alveolar air. E - How does the Breathalyzer™ work? F - Redox chemistry inside the Breathalyzer™ G - The Breathalyzer™ assumes a specific blood-to-breath-ratio to calculate the BAC 4A - How does ethanol get to the lungs? Ethanol must first enter the bloodstream Ethanol is a water soluble compound—it dissolves readily in water-based solutions and once it is swallowed, it moves into water spaces throughout the body, including the bloodstream. Here is a brief review of what happens: Molecules of ethanol travel across the membranes made of epithelial cells that line the stomach and small intestine. Ethanol molecules move across the membrane via passive diffusion. Once on the other side of the gut cells, these small ethanol molecules then easily pass through the walls of the tiny capillaries that line the gut. To review the details about ethanol absorption go to Module 1B. Learn more about diffusion across a biological membrane. Figure 4.1 Absorption of ethanol from the gut into the bloodstream. Ethanol molecules in the gut diffuse passively across epithelial cells, through the interstitial space, and then into nearby capillaries. Ethanol travels through the bloodstream to the lungs Ethanol, in its liquid form, travels in the capillaries to the veins, and then heads up to the lungs. It takes the same route that deoxygenated blood takes to become oxygenated. Refer to Figure 4.2 to follow the route that the ethanol takes to the lungs. Ethanol is carried by the venous circulation to the right side of the heart. Ethanol enters the right atrium (like a foyer), and then moves into the right ventricle (a pump). The right ventricle pumps the venous blood to the lungs. In the lungs, red blood cells unload carbon dioxide and pick up oxygen brought in by respiration (breathing). Some of the ethanol dissolved in the blood gets eliminated as a gas by the lungs during exhalation. Ethanol that remains in the blood returns to the left side of the heart (the atrium). The left atrium supplies blood to the left ventricle of the heart, which pumps the blood to the rest of the water-containing compartments and tissues of the body via the arteries. The branch of the circulatory system that moves blood between the heart and the lungs is called the pulmonary circulation, while the branch that moves blood throughout the rest of the body is called the systemic circulation. Figure 4.2 Ingested ethanol travels from the gut to the liver before reaching the heart and lungs. Inhaled ethanol travels to the brain before it goes to the liver. Ethanol in the veins travels to the heart, then the lungs, where a portion is exhaled. The arterial side is red, the venous side is blue. Adapted from www.thepepproject.net. Ethanol moves passively across membranes in the gut into the capillaries, bringing it to the veins, leading to the heart. From there it goes to the lungs where a small portion is excreted. Most of the ethanol goes back to the heart and then to the arteries. 4B - Ethanol vaporizes to a gas in the lungs In the lungs, ethanol moves by passive diffusion from the capillaries into the surrounding air sacs called alveoli. Unlike the process of active transport, passive diffusion requires no additional energy to move molecules. The energy comes from the difference in the concentration of ethanol between the two compartments: blood and lung alveoli (substances move from areas of high concentration to areas of low concentration). As ethanol diffuses out of the pulmonary capillaries, it contacts a thin water-based mucous layer on the inner surface of the alveolar sacs (Figure 4.3). It is inside the alveolar sacs of the lung—at the interface between the liquid mucous layer and the air—where some of the ethanol changes from a liquid into a gas. The process is known as vaporization. Figure 4.3 Ethanol from the heart reaches the lung capillaries (purple), which surround the alveoli. As ethanol diffuses into the alveoli, it vaporizes to a gas. Learn more about the structure and function of the lungs. How does ethanol vaporize into a gas? Let's look more closely at the process of ethanol vaporization. At room and body temperatures, ethanol can exist as a liquid or a gas. Liquid - individual ethanol molecules are tethered to one another by two types of chemical forces: hydrogen bonds and Van der Waals forces. Gas - when liquid ethanol molecules have enough energy to break the hydrogen bonds connecting them together, they escape into the gas state. Compounds, like ethanol, that can easily change from a liquid to gas are called volatile compounds. Volatility is related to the number of hydrogen bonds. Let's compare ethanol and water. Individual ethanol molecules can only form three hydrogen bonds with neighboring ethanol molecules, while water molecules can form as many as four hydrogen bonds with neighboring water molecules. So, ethanol requires less energy to break three hydrogen bonds (ethanol) than four hydrogen bonds (water). Therefore, ethanol is more volatile than water. Figure 4.4 Ethanol vaporizes when its hydrogen bonds are broken. This happens more easily compared to water because ethanol molecules are bound to each other with up to three hydrogen bonds, while water molecules are bound to each other with up to four hydrogen bonds. Learn more about vaporization and the volatility of a compound. Ethanol volatilizes in the alveoli easily; ethanol has less hydrogen bonds to break compared to water in order to transition from a liquid to a gas. 4C - Ethanol leaves the lungs in the air As you breathe in, oxygen from the atmosphere enters the lungs and moves into the circulation through the capillary walls that surround the alveoli of the lung. When you exhale, both carbon dioxide and ethanol in the breath are removed from the blood (of course, not all of it!). Let's look at this more closely. Ethanol dissolved in the mucous lining of the alveoli is in the liquid phase. Above the mucous layer is open space. As the ethanol vaporizes, the gas (vapor) expands to fill the air space within the alveolar sac. When you exhale, the vaporized ethanol exits the body. This is the same ethanol vapor that is measured by the Breathalyzer™ test to determine whether someone is intoxicated. So, respiration not only provides the body a steady supply of oxygen, but also provides an exit for toxic waste products, including some volatile drugs like ethanol. Figure 4.5 As ethanol diffuses into the alveoli, it vaporizes to a gas. Ethanol vapor can be exhaled from the body. Ethanol can leave the body through exhalation; once the ethanol moves from the liquid to gas phase in the alveoli, it now has an open route to the external environment. Very little alcohol leaves the body through this route, but it’s easy to smell it! 4D - Ethanol in the blood equilibrates with ethanol in the alveolar air. The ability of ethanol to volatilize in the lungs makes it a good candidate to be detected with breathing devices such as the Breathalyzer™. The Breathalyzer™ test approximates a person's blood alcohol concentration (BAC) because the concentration of alcohol vapor in the lungs is directly related to its concentration in the blood. Although several devices can be used to test for alcohol in the breath, each one is based on a unique chemical property of alcohol and all are based on the central scientific principle that alcohol in the blood is in equilibrium with alcohol in the expired air. How is equilibrium established? Shortly after the first sip of beer, the concentration of ethanol in the blood exceeds the concentration of ethanol in the alveoli (which is zero). Ethanol diffuses into the air sacs until the concentration in blood relative to that in air achieves a constant ratio. Now, the two compartments are in equilibrium with one another. During equilibrium, molecules of ethanol may continue to move freely back and forth between the blood and the alveoli. But for each molecule that moves from the air sac back to the blood, another molecule moves from the blood to the air sac; so, no net movement of ethanol occurs across the membrane. This equilibrium is dynamic because it can react to disturbances within either compartment. For example, when ethanol in the alveoli leaves the body in the breath, the ratio of ethanol concentration between the blood and the alveoli changes. To restore the ratio, more ethanol moves from the blood into the air sacs until the net flow of ethanol between the compartments is zero--equilibrium is restored. Figure 4.6 After drinking ethanol, it equilibrates between blood and the alveolus (left panel). With each exhalation, the equilibrium is disturbed, and more ethanol moves into the alveolus to reestablish the equilibrium (right panel). The validity of a Breathalyzer™ test is dependent on the equilibrium of ethanol in the blood and the alveolar air. No net movement of ethanol across the alveolar membrane occurs at equilibrium. 4E - How does the Breathalyzer™ work? The original version of the Breathalyzer™ included a mouthpiece and two chambers containing liquid connected to a meter that detects a change in color. To use the Breathalyzer™, the subject exhales through the mouthpiece into a test chamber filled with a reddish-orange solution of potassium dichromate (K2Cr2O7). Figure 4.7 An example of a newer version of a breath testing device In the Breathalyzer™, alcohol reacts with the reddish-orange potassium dichromate solution and turns green. The degree of the color change is directly related to the level of alcohol in the expelled air. A photocell compares the difference in colors between the reacted mixture in the test chamber and a reference chamber containing unreacted mixture. The difference in colors produces an electrical current, which can be converted into a quantitative value for the BAC. Read on to understand how the chemical reaction produces the color change. Figure 4.8 The ethanol vapor in the Breathalyzer™ triggers a chemical reaction with the compounds packed inside, turning the chemicals from orange to green. The more ethanol present, the greater the amount of green color is produced. A description of different types of breath analyzers and how they work can be found at http://science.howstuffworks.com/breathalyzer3.htm The original Breathalyzer™ devices relied on the color change of a solution of potassium dichromate from red-orange to green. The more alcohol in the breath, the more the color changes to green, and this is quantified by the device. 4F - Redox chemistry inside the Breathalyzer™ The chemical reaction inside the Breathalyzer™ includes both oxidation and reduction. The Breathalyzer™ contains a chamber with several compounds to support these reactions. They include: Potassium dichromate Sulfuric acid Silver nitrate When the potassium dichromate solution in the Breathalyzer™ reacts with ethanol, the potassium dichromate loses an oxygen atom. This process is called reduction --when a compound loses oxygen, gains hydrogen, or gains (partially gains) electrons. The reduction converts orange potassium dichromate into a green solution containing chromium sulfate. At the same time dichromate ion gets reduced to chromium ion, ethanol gets oxidized to acetic acid. Oxidation reactions often occur simultaneously with reduction reactions and are commonly abbreviated as redox reactions. Oxidation occurs when an element combines with oxygen to give an oxide. For example, the oxide of hydrogen is water. Oxidation is the gain of oxygen, the loss of hydrogen, or the loss (or partial loss) of electrons Silver nitrate serves as a catalyst for the reaction to increase the rate at which the dichromate gets reduced. Sulfuric acid in the test chamber helps to remove the alcohol from the exhaled air into the test solution and to provide the necessary acidic conditions. Oxidation and reduction (redox) reactions are opposing reactions that occur simultaneously. Figure 4.9 This is the balanced equation to show how ethanol is oxidized to acetic acid Learn more about redox reactions. In the Breathalyzer™, ethanol participates in a redox reaction; it gets oxidized as it loses electrons (H atoms) and the potassium dichromate gets reduced as it picks up some electrons. 4G - The Breathalyzer™ assumes a specific blood-to-breath-ratio to calculate the BAC In a breath test, one must calculate how much ethanol measured from the expired air sample is in the blood. To do this, one must take into account the volume of blood from which the ethanol originated. First, a few assumptions must be made: 1. The ethanol concentration in the expired alveolar air is directly proportional to the ethanol concentration in the blood. This assumption is based on Henry's Law which states that, at constant temperature, the concentration of gas dissolved in a liquid is proportional to its concentration in the air directly above the liquid. 2. The exhaled alveolar air is in equilibrium with the blood. In the case of ethanol, the ratio of concentrations of ethanol in blood to expired alveolar air has achieved a constant value—on average, this is 2100:1. This relationship is called the blood-to-breath ratio or partition ratio. The blood-to-breath ratio can vary between 1500:1 and 3000:1. The ratio varies: among individuals according to age, gender, genetic makeup, and state of intoxication within a given individual at different times among measuring devices The 2100:1 ratio is used as a standard conversion factor in determining BAC from the Breathalyzer™ test. It is actually an average based on comparisons of blood and breath samples collected from many individuals using several types of measuring devices. The variability in the inter-person and intra-person blood-to-breath ratio could under or overestimate the true BAC as measured directly from the blood. This could be acceptable in court if there is proof of a discrepancy. The average blood-to-breath ratio is about 2100:1. This means that 1 milliliter of blood has 2100 times more ethanol than 1 milliliter of air from the lungs. This value is used to calculate the BAC from the Breathalyzer™ test. Module 4 - Student Quiz 1) Where does carbon dioxide-rich blood from the capillaries in the gut go? A. lungs → right side of the heart → left side of the heart B. right side of the heart → the lungs → left side of the heart C. left side of the heart → the lungs → right side of the heart D. lungs → left side of the heart → right side of the heart 2) Ethanol is more volatile than water because it has: A. fewer hydrogen bonds B. more hydrogen bonds C. carbon atoms D. nitrogen atoms 3) Which of the following statements describes ethanol equilibrium between the blood and the alveoli? A. The amount of ethanol is the same in the blood relative to the alveoli. B. The concentration of ethanol is the same in the blood relative to the alveoli. C. Ethanol stops moving between the blood and the alveoli. D. There is no net flow of ethanol between the blood and the alveoli. 4) No energy is required for the passive diffusion of ethanol out of the gut and into the bloodstream. If energy is not required, then what drives the diffusion process? A. ethanol's small molecular size B. proton pumps C. the higher concentration of ethanol in the bloodstream relative to the gut D. the higher concentration of ethanol in the gut relative to the bloodstream 5) When hydrogen is burned it combines with oxygen to produce water molecules. Which describes the state of each atom? A. Hydrogen is oxidized, Oxygen is reduced B. Oxygen is oxidized, Hydrogen is reduced C. Both are oxidized D. Both are reduced MODULE 4 - ACTIVITY 4A - The Breathalyzer™ Isn't Always Right! In the Spritney Beers case, the reported BAC was 0.08%, based on the 2100:1 blood-to-breath ratio. If her blood-to-breath ratio was actually closer to 1500:1, would the calculated BAC of 0.08% be an underestimate or an overestimate—by how much? Her BAC calculation would be underestimated by a factor of: 2100/1500 = 1.4 So, taking into account the underestimation, the real BAC would be: 0.08 x 1.4 = 0.112% If her blood-to-breath ratio was closer to 3000:1, would the calculated BAC of 0.08% be an underestimate or an overestimate—by how much? Her BAC calculation would be overestimated by a factor of: 2100/3000 = 0.7 So, taking into account the overestimation, the real BAC would be: 0.08 x 0.7 = 0.056% Thus, according to these estimates, the true BAC could range from of 0.056% to 0.112%. In most states 0.056% is under the legal limit (0.08%) for intoxication. So it's possible she could have been under the limit, but in our legal system, she would have had to prove that in order to have the case dropped. Her only way to prove it would have been to submit to a blood test immediately after the breath test. MODULE 4 - RESOURCES Books and Articles Burke, Barbara A. Journal of College Science Teaching. Measuring Breath Alcohol Concentrations. Washington: Dec 1991. Vol 21, Issue 3; p 176. Web-based References Alcohol, Chemistry and You: a description of Fick's law as it describes the passive movement of alcohol across a biological membrane. www.chemcases.com/alcohol/alc-04.htm Nice explanation of the chemistry behind the breathalyzer http://science.howstuffworks.com/breathalyzer.htm Basis of the partition coefficient: science and law http://www.forensicevidence.com/site/Biol_Evid/Breath_Tests.html Videos and animations Video demonstrating Le Chatelier's principle using the chromate-dichromate equilibrium http://jchemed.chem.wisc.edu/JCESoft/CCA/CCA8/MAIN/8/08/32/thumbs.html Classroom Curricula and Activities Lab Exercise (for advanced chemistry class): Breathalyzer Reaction in the Lab www.chem.byu.edu/Plone/coursematerials/exploratorylab/EL64.PDF/view http://educ.queensu.ca/~science/main/concept/chem/c03/C03deae4.htm This exercise may work best as a teacher demonstration due to the hazardous nature of the chemicals involved. This lab would allow students to practice with a concentration-response curve. Add increasing concentrations of ethanol to the reaction. Use a spectrophotometer if available to measure absorbance of the colored solution compared with the control. Try mouthwash containing trace amounts of alcohol. This reaction should be sensitive enough to pick up the alcohol. Practice Problem: Balancing Chemical Equations – The Breathalyzer Test http://chemed.chem.purdue.edu/genchem/topicreview/bp/ch19/problems/ex19_3s.html Practice with Redox reactions and oxidation states of molecules