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- In vitro experiments are conducted at pH = 7.4 to simulate physiological conditions. A phosphate buffer system is often used. H₂PO → H₂PO¹²¯ +H+ pK₁ = 7.2 a. What must be the ratio of the concentrations of HPO to H₂POд ions? b. What mass of NaH2PO4 must be added to 500.0 mL of 0.10 M Na₂HPO4 (aq) in the preparation of the buffered solution?7. · In the space provided, sketch a titration curve (pH vs H+ ions dissociated) of a phosphate buffer shown below. The pKa of H2PO4 is 7.0. H,PO, НРО, + 9. 8. 7 pH 3 1.0 0.5 H+ dissociated --> b. What is the effective buffering range of this acid? At what pH will you have 25% of the buffer in the form of H2PO4? с.You have been provided with stock solutions of: stock A: 0.06 M sodium pyrophosphate buffer pH 8.5 stock B: 3 M ethanol stock C: 0.015 M NAD+ stock D: milli Q water Determine the volume you will need of each solution to prepare a buffer of with a final volume of 60 mL containing 10 mM sodium pyrophosphate pH 8.5, 100 mM ethanol, 1 mM NAD+. i.e. volume of stock A = _________mL volume of stock B = _________mL volume of stock C = _________mL volume of stock D = _________mL Show your calculations to arrive at your answers.
- Phosphate buffered saline (PBS) is a physiological buffer often used in biology experiments. Because the concentrations are very low and small amounts are difficult to weigh out, it is often prepared as a 10X stock solution meaning that it is prepared at 10 times the working concentration which is 1X. 1X PBS is made up of Na2HPO4 10 mM (MW 141.9 g.mol-1); KH2PO4 8 mM (MW 136.08 g.mol-1), NaCl 137 mM (MW 58.44 g.mol-1) and KCl 74.5 mM (MW 74.5 g.mol-1). You decide to prepare a 1 L 10X solution of PBS. What is the Molar (M) concentration of each of the components in you 10X stock. Write your answers in the table below. How much of each of the components should you weigh out to prepare 1L of the 10X stock solutions. Write your answers in the table below.1. Calculate the pH of the following solutions: A. [H+] = 1.36 x 10-9 M В. [ОН] —D 3.86 x 10-8 М 2. Calculate the pOH of the following solutions: A. [OH-] = 8.6 x 10-8 M В. рH %3D 3.2 3. What is the pH of a buffer prepared by mixing 100 ml 0.050 mM NAH2PO4 and 25 ml 0.075 mM Na2HPO4? (рКa1%32.2; рКо23 7.21; pКаз312.7)Acetazolamide is a drug which inhibits carbonic anhydrase. Carbonic anhydrase participates in regulation of the pH and bicarbonate content of a number of body fluids. Figure 2 shows the experimental curve of initial reaction velocity (as percentage of Vma) versus [S] (concentration) for the carbonic anhydrase reaction. The graph also shows the curve in the presence of acetazolamide. 100 No inhibitor 50 Acetazolamide 0.2 0.4 0.6 0.8 [S] (mM) Figure 2 (i) Compare the maximal velocities and Michaelis Menten constants of the enzyme in the absence and the presence of the inhibitor acetazolamide. Determine the nature of inhibition by acetazolamide. Explain your answer. (ii) Name TWO (2) other types of inhibitions besides the inhibition shown by acetazolamide. Sketch a graph of V versus [S] showing curves in the absence of an inhibitor and in the presence of the types of inhibitors not shown by acetazolamide. ("AJO %) A
- 1. What volumes of 50 mM CH3COOH and 30 mM NaCH3CO0 solutions are needed to prepare 2L of 20 mM acetate buffer pH 3.6? (pka for acetic acid is 4.74)Acetazolamide is a drug which inhibits carbonic anhydrase. Carbonic anhydrase participates in regulation of the pH and bicarbonate content of a number of body fluids. Figure 2 shows the experimental curve of initial reaction velocity (as percentage of Vmax) versus [S] (concentration) for the carbonic anhydrase reaction. The graph also shows the curve in the presence of acetazolamide. 100 No inhibitor 50 Acetazolamide 0.2 0.4 0.6 0.8 (S] (mM) Figure 2 (i) Compare the maximal velocities and Michaelis Menten constants of the enzyme in the absence and the presence of the inhibitor acetazolamide. Determine the nature of inhibition by acetazolamide. Explain your answer. (*"A JO %) A1. What is the molality of the following solutions? a. 171.9 g of Sr(NO3)2 is dissolved in 1.44 kg of water. b. 0.883 g of K3PO4 is dissolved in 40.0 g of water. 0.250M=0.250mcl kF IL scl i80.anilio (6250 mcl 0.500M=0.500md wO IL Som
- Rp Dextrose hydrous 50.0 g Sodium Chloride 9.0 g Calcium Chloride 0.3 g WFI (Water for injection) qs… 1 L Calcium Chloride MW: 111; Sodium Chloride MW: 58.5; Dextrose MW: 198 a. Please calculate how many mOsmol/L is the above LVP solution? b. How many mOsm/L Ca++ and Na+ contain the above solution? c. What is the above LVP solution tonicity?3. Show that the inverse of equation 29-78 in the textbook gives the Lineweaver-Burk Equation. The hydration of CO2 is catalyzed by the enzyme carbonic anhydrase. For a total enzyme concentration of 2.32 x 10-⁹M, the following data were obtained. [CO2]o (M) v (M/s) 0.00125 2.78 x 10-5 0.00250 5.00 x 10-5 0.00500 8.33 x 10-5 0.02000 1.66 x 10-4 Plot these according to the Burke-Lineweaver Equation and determine the values of Km, the Michaelis constant, and k2, the rate constant for product formation from the enzyme-substrate complex from the slope and intercept of the plotted data.3. Compute for the amount of each component of KCN broth if you were to prepare 280 ml. Express your answers using two decimal places. Follow this format in typing your answers: answerabbreviated unit (e.g. 3.00 g, 0.75 ml, 10.25 mg): Components Amount per liter Polypeptone 3.0 g N22HPO4 5.64 g (Available as NazHPO4 · 7H2O*) Monopotassium phosphate 0.225 g Sodium chloride 0.5% KCN 0.075 g (Available as 10,000 ppm solution) *MW (g/mol): Na-23; P-31; H-1; 0-16 3.3. Monopotassium phosphate /r