BENEDICT’S TEST: 1.) What color does a solution containing Benedict’s reagent and placed in hot water turn in the presence of glucose? (10pts) ______________________________ 2.) How do you think the results of the Benedict’s test would differ if you used it on a solution that was 1% glucose vs. a solution that was 0.2% glucose? (20pts) 3.) In the video about the Benedict’s test, what was the point of having the test tube with just water? DO NOT use the word CONTROL – explain it in your own words. (20pts) 4.) According to the lab manual, how much Benedict’s reagent are you supposed to add to the test tube? (10pts) ___________________________________ 5.) According to the lab manual, how much candidate solution are you supposed to add to the test tube? (10pts) ___________________________________ 6.) For the clean up of the Benedict’s test, where are you supposed to dispose of your Benedict’s waste? (10pts)

BARIUM HYDROXIDE TEST FOR CO­ 2 : 7.) When you bubble CO 2 gas into a solution of Barium Hydroxide, what will visually indicate a reaction of CO 2 with Ba(OH) 2. Hint: the word starts with a ‘P’ (10pts) 8.) According to the lab manual, how much Barium Hydroxide will you need to add to the test tube? (10pts) ________________________________ BONUS: What is one of the learning goals (Victory Conditions) of today’s lab (Lab 10 – Glycolysis and Fermentation)? (10pts)

Glycolysis and Fermentation What’s so sweet about Sugar?

Objectives Discuss ATP and where the ‘energy’ is Describe what fermentation is and where the ‘energy’ is Design an experiment to ‘test’ fermentation Weigh the merits of fermentation vs. respiration Recognize/explain what happens in a RedOx reaction

Thinking it Through CH 4 (methane) C 2 H 5 OH H 3 C-CH 2 -OH (ethanol) C 8 H 18 CH 3 (CH 2 ) 6 CH 3 (octane) H 2 O CO 2 Similarities in components/structure? Differences in behavior? Differences in structure?

So where are ‘calories’? Energy (heat) needed to raise 1g water, 1 o C Saturated Fat Why are there more calories?

RedOx Reactions

OIL RIG OIL = Oxidation Is Loss RIG = Reduction Is Gain of electrons

O

Nicotinamide adenine dinucleotide

NAD+

Nicotinamide adenine dinucleotide NAD+

H H H

Oxidized Form Reduced Form

Oxidized Form Reduced Form NAD+ NADH

“ATP- molecule used to store and transfer energy”

Phosphate Groups

“ATP- molecule used to store and transfer energy” Why not just separate? What does this mean in terms of ‘energy’?

How does breaking a bond ‘release’ energy? Same thing shown with a bit more detail…

Metabolic Pathways

Glycolysis Glyco – glucose/sugar Lysis – “to loosen”, take apart

Glucose (6 Carbons) Glycolysis

Glucose (6 Carbons) 2 ATP 2 ADP Glycolysis

Glucose (6 Carbons) 2 ATP 2 ADP 2 NAD+ Glycolysis

Glucose (6 Carbons) 2 ATP 2 ADP 2 NADH 2 NAD+ Glycolysis

Glucose (6 Carbons) 2 ATP 2 ADP 2 NADH 2 NAD+ Glycolysis NAD+ is reduced to “NADH”

Glucose (6 Carbons) 2 ATP 2 ADP 2 NADH 2 NAD+ Glycolysis *Addition of 2 inorganic phosphates

Glucose (6 Carbons) 2 ATP 2 ADP 2 ATP 2 ADP 2 NADH 2 NAD+ Glycolysis

Glucose (6 Carbons) 2 ATP 2 ADP Pyruvate 3 Carbons Pyruvate 3 Carbons Pyruvate 3 Carbons 2 ATP 2 ADP 2 NADH 2 NAD+ Glycolysis

Glucose (6 Carbons) 2 ATP 2 ADP Pyruvate 3 Carbons Pyruvate 3 Carbons Pyruvate 3 Carbons 2 ATP 2 ADP 2 NADH 2 NAD+ Glycolysis

Pyruvate 3 Carbons Net production of 2 ATP Glycolysis Glucose (6 Carbons) 2 ATP 2 ADP Pyruvate 3 Carbons Pyruvate 3 Carbons 2 ATP 2 ADP 2 NADH 2 NAD+

How do you Keep Glycolysis Going?

Glucose (6 Carbons) 2 ATP 2 ADP Pyruvate 3 Carbons Pyruvate 3 Carbons 2 ATP 2 ADP 2 NADH 2 NAD+

How do you Keep Glycolysis Going? Glucose (6 Carbons) 2 ATP 2 ADP Pyruvate 3 Carbons Pyruvate 3 Carbons 2 ATP 2 ADP 2 NADH 2 NAD+

Fermentation (Lactic Acid) Lactic Acid 3 Carbons Pyruvate 3 Carbons 2 NAD+ 2 NADH NADH is oxidized into NAD+ Pyruvate is reduced and forms Lactic Acid

Fermentation (Alcoholic) Ethanol 2 Carbons Pyruvate 3 Carbons 2 NAD+ 2 NADH NADH is oxidized into NAD+ Pyruvate is reduced and forms Ethanol CO 2

Fermentation Keep Glycolysis going - oxidizes NADH into NAD+ Glycolysis – 2 ATP “NO” O 2 present Lactic Acid Alcoholic –Ethanol - CO2

Here’s the claim: Glycolysis – Glucose + 2ATP  Pyruvate + 4ATP Fermentation (no O 2 present) Pyruvate + NADH  ethanol + CO 2 + NAD + What can you test? Conditions needed? How? Rank in terms of: feasibility, safety, ‘fun’

Optimizations We’ve proposed testing for components Are all tests best done under same conditions? Concept: limiting components. How do you max your yield (or sensitivity) for: –Glucose –CO 2 –ETOH

More? What will your solution look like? –“murky”? How do you speed it up? What conditions do we WANT?

Time to get to know each other better…

Okay, that’s Fermentation How does it ‘usually’ happen?

So? WHY do you breath O 2 ? Where does the CO 2 come from? Where does ‘body heat’ come from? What is the relationship between sugars, and fats?

What do ‘conclusive’ results tell you? Smoking, etc. –‘consistent’ with