CB Workshop 1 answers - 2007

Clinical Biochemistry Workshop 1 Basic Laboratory calculations

1. To make the following saline solution (5 litres 0.9% w/v sodium

chloride) what would you weigh out? 1% solution = 1g in 100 ml 0.9% soln = 0.9g in 100ml Therefore 9g in 1 litre and 5 x 9g in 5 litres = 45g NaCl 2. The atomic weights of Na and Cl are 23 and 35 respectively. What is the concentration in mmol/l of the solution from question 1? Mw NaCl = 58 Therefore 1M or 1mol/l solution = 58g in 1 litre From question 1 = 9g in 1 litre So, 9/58 = 0.155mol/l = 155 mmol/l What is the concentration in mol/ml? 1mmol = 1000 mol 1litre = 1000ml So, 155 mmol/l is the same as 155 mol/ml 3. If you took out 100ml of the solution from question 1 a) What would the concentration of NaCl (mmol/l) be in that sample? In the removed 100ml the concentration is the same = 155 mmol/l CONCENTRATION is the same AMOUNT is different 155mmol/l, so in 100ml = 155 / 10 = 15.5 mmol or 15, 500 mol Total = 15.5 mmol Mw NaCl = 58 1mol = 58g NaCl, Remember 1mol = 103 mmol
b) How many moles would the sample contain?

c) How many mg NaCl would the sample in 3b contain?

CB Workshop 1 answers - 2007

Then 15.5 mmol = 58 x 15.5 x 10-3 = 0.899g or 899 mg

d) How many g NaCl would the sample in 3b contain?

= 0.899g see above 4. What would the concentration ( mol/ml) be if you diluted a 600mmol/l solution as follows a) 100-fold b) 1:100 c) 1+99 d) 10-fold followed by 10-fold Some of the above are the same 100-fold = 1ml in 100ml total volume 1:100 = 1ml plus 100ml 1+99 = 1ml in 100ml total volume 10-fold followed by 10-fold = 1ml plus 9ml. Remove 1ml of this solution and add 9ml water Remember that 600 mmol/l is the same as 600 mol/ml 4a) 100-fold 100ml sample = 600 mol/ml, therefore diluting 1ml x 100 = 600 / 100 = 6 mol/ml 4b) 1:100 1ml plus 100ml water, so total volume = 101ml. 1ml = 600 mol/ml = 600 / 101 = 5.94 mol/ml 4c) 1+99ml 1ml plus 99ml, so total volume = 100ml 1ml = 600 mol/ml = 600 / 100 = 6 mol/ml 4d) 10-fold followed by 10-fold 1ml plus 9ml, = 60 mol/ml. Take 1ml of this and add to 9ml = 6 mol/ml 5 a) If you have a solution containing 1mmol/l how would you prepare 10ml solutions of 900 mol/l, 600 mol/l or 300 mol/l?

CB Workshop 1 answers - 2007

1mol/l = 1000 mol/l To prepare 900 mol/l need to dilute 1000/900 = 1.11 fold. In 10 ml = 10/1.11 = 9.0ml 9.0 ml stock solution plus 1ml water = 900 mol/l To prepare 600 mol/l need to dilute 1000/600 = 1.66 fold. In 10 ml = 10/1.66 = 6ml 6ml stock solution plus 4ml water = 600 mol/l To prepare 300 mol/l need to dilute 1000/300 = 3.33 fold. In 10 ml = 10/3.33 = 3ml 3ml stock solution plus 7ml water = 300 mol/l b) If you have a solution containing 1mmol/l how would you prepare 5ml solutions of 900nmol/l, 600nmol/l or 300nmol/l? Remember 1mmol = 1000 mol and 1 mol = 1000nmol. So, dilute all solutions 1000 fold first then repeat steps in 5a). Dilute 1mmol/l stock 1000-fold = 100-fold followed by 100-fold Repeat all steps as above with diluted 1000 mol stock c) If you mix together 25ml of a solution containing 700 mol/l with 25ml of a solution containing 500 mol/l, what would the concentration be ( mol/l)? Concentrations are additive, but remember that you also double the volume. So, 700 mol/l plus 500 mol/l = 1200 mol/2 litres Therefore 600 mol/l 6. The coenzyme NADH has a molar extinction coefficient of 6220 at a wavelength of 340nm. a) What would be the absorbance of a solution containing 35 mol of NADH in 1 litre? Molar extinction coefficient is the absorbance for 1M solution So, 1mmol/l or 1mM solution = 1000 times lower concentration = 6220 / 1000 = 6.22 35 mol/l = 1000 / 35 = 28.6 times lower concentration 6.22/28.6 = 0.217

CB Workshop 1 answers - 2007

b) What would be the absorbance of a solution containing 35 mol of NADH in 5 litres? 35 mol/l = 0.217 In 5 litres = 0.217 / 5 = 0.043 c) If the solution (35 mol of NADH in 5 litres) was diluted 5-fold, what would its absorbance be? If the solution from 6b) was diluted 2-fold, the absorbance would be 5-fold lower = 0.043 / 5 = 0.0087 d) What is the NADH concentration (in mol/l) of a solution with an absorbance of 0.76? Using Beer-Lambert law A= cl A = absorbance = molar extinction coefficient C = concentration (M) L = pathlength of cuvette (usually 1cm) We know the extinction coefficient and the absorbance, so we can calculate the concentration: A= cl 0.76 = 6220 x c x 1 0.76 / 6220 = c = 1.22 x 10-4M (moles / litre) = 122 mol/l e) What is the concentration of the solution in 6d) in mol/ml? 1M = 1x106 M 122 mol/l Therefore 122 / 1000 = 0.122 mol/ml f) If the molecular weight of NADH is 709, how many grams are there in solution 6d)? 1M solution of NADH = 709g/l Solution from 6d) = 709 x 1.22x10-4 = 0.086g

CB Workshop 1 answers - 2007

7. A standard solution of picric acid (10mmol/l) gives an absorbance of 0.86. a) What is the concentration (mmol/l) of a solution with an absorbance of 0.39? The relationship between concentration and absorbance is linear in this range. This means they are proportional. A = 0.39 is less than 0.86 0.86 / 0.39 = 2.2 times lower Thus concentration is also 2.2 x less = 10 / 2.2 = 4.55mmol/l b) If 1.0ml of the 10mmol/l solution was mixed with 19.0ml of water, what would the absorbance be? 1.0ml 10mmol/l stock plus 19ml water = 20fold dilution = 10 / 20 = 0.5mmol/l 10mmol/l absorbance = 0.86 Therefore 20-fold less = 0.86 / 20 = 0.043 c) If a portion of the diluted solution 7b) were mixed with an equal volume of picric acid (10mmol/l), what would be the concentration? 0.5mmol/l plus 10mmol/l Added together = 10.5 mol in 2 litres Therefore 5.25mmol/l d) What would be the absorbance of the solution in 7c)? 10mmol/l picric acid solution A = 0.76 Therefore 5.25mmol/l solution = 10 /5.25 = 1.9 times less = 0.76 / 1.9 = 0.4