Basic Laboratory Mathematics

Self-Instructional Material

SIGNIFICANT FIGURES Those figures in a number that have meaning. The decimal format is used in reporting significant figures. In the laboratory setting, significant figures are determined by the precision and accuracy of a given protocol or piece of equipment. Example: A car speedometer You can't say your speed is 55.467 mph with current speedometers. Car speedometers are accurate + 0.5 mph for analog (dial with needle) meters; therefore a speed of 55 mph is really 54.5 to 55.5 mph. While it is possible to calculate out several places, the information is meaningless if it is beyond the accuracy of the procedure. This is equivalent to false magnification on a microscope where no further resolution is available regardless of the increase in magnification. Rules for recording significant figures: 1. All non-zero digits are significant 2. All zeros between non-zero digits are significant 3. Zeros to the left of implied decimal point may or may not be significant; (one convention uses a bar over the last significant zero, i.e 1200) 4. Zeros to the right of a decimal point and a non-zero digit are significant; they indicate the precision of a number. 5. Zeros to the left of non-zero digit and to right of decimal point are not significant if the value of the number is less than one. Overall the significant figure represents the accuracy and precision of the procedure that produced the number. Example: Test value is reported as 12.6. This implies that the procedure is accurate to the nearest tenth and the actual value is between 12.55 and 12.65. Note that 12.60 implies accuracy to nearest hundredth with the actual value between 12.595 - 12.605. Rounding Off Removing insignificant digits produced as a result of computations. This is done to prevent an implied accuracy due to presence of insignificant digits.

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GENERAL RULES FOR ROUNDING OFF 1. If digit dropped is less than 5 then there is no change to preceding digit. 12.64 rounds to 12.6 2. If digit dropped is greater than 5 then the preceding digit is increased by one. 12.66 becomes 12.7 3. If digit dropped is 5 then add 1 if preceding digit is odd, add 0 if preceding digit is even. 3.15 becomes 3.2 3.25 becomes 3.2 Figure Retention When working with figures that have various levels of significant figures, retain only as many significant figures in data as will be necessary to give only one uncertain figure. Example: Reading pipet or buret for titration. The scale is in 0.l ml increments which means you can estimate values between scale divisions (this is called interpolation). Therefore you can report 3.62 ml of acid used; 3.6 is scale reading, .02 is the interpolation. The number 3.62 has only one uncertain figure. Note 3.62 implies that the actual value is between 3.615 & 3.625. This would only be the case if you felt that your estimate was that accurate. If you doubt your ability to be that accurate then report the value as then 3.6. This implies an accuracy of 3.55 to 3.65ml. In general, when working with numbers with different levels of significant figures, all numbers are rounded to the accuracy of that number with least number of significant figures (i.e. the least accurate number). Addition/Subtraction When adding or subtracting numbers with differing numbers of significant figures, keep only that number of significant figures in each number equal to that of the number with the fewest significant figures occurring after the decimal point. Record the answer with that same number of significant figures. 0.0212 + 29.64 + 1.056931 The fewest significant figures after decimal point is 2. 0.0212 rounds to 0.02 29.64 rounds to 29.64 1.056931 rounds to 1.06 30.72 Multiplication/Division When multiplying/dividing figures with differing numbers of significant digits consider the total number of significant figures, not just those occurring after the decimal point. 0.0211 x 25.63 x 1.05881 0.0211 x 25.6 x 1.06 = 0.573 (0.5725696)

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EXPONENTS The number to be multiplied is the Base; times to be multiplied is power/exponent. For laboratory work use two different bases: 10-metric Metric 102 = 10x10 = 100 Base two 22 = 2x2 = 4 Negative exponent indicates the reciprocal of the base is multiplied 102 = 10x10 -2 10 = 1/10x1/10 = 1/100 2-2 = 1/2x1/2 = 1/4 When multiplying numbers with the same base, add exponents 102 x 103 = 105 102 x 10-3 = 10-1 (100/1) x (1/1000) = 1/10 When raising to Higher Power, Multiply Exponents (102)3 = 106 (not 105) When manipulating numbers with different bases, convert to corresponding simple non-exponent numbers first. 23 x 102 = (2x2x2) x (10x10) = 8 x 100 = 800 23 + 102 = (2x2x2) + (10x10) = 8+100 = 108 Scientific notation is the use of base 10 exponents to express very large/small numbers. Format: Two parts multiplied together; first part contains significant figures, second part indicates size: 100 = 1x102 278 = 2.78x102 376,000 = 3.76 x 105 NOTE: a number raised to the 0 power equals 1: 20 = 1, 100 = 1 Converting to Scientific Notation Numbers greater than 1 Count number of places required to move decimal point to yield number with one digit to left of decimal point; add 1 to the exponent for each place: 125 = 1.25x 102 Numbers less than 1 Count number of places necessary to move decimal point to right of first non-zero digit; subtract 1 from the exponent for each place: 0.0012 = 1.2x10-3 Calculations To add/subtract convert to either simple number or same exponent whichever is easier: 102 + 103 = 100+1000 = 1100
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2-serial dilution

(1.2 x 102) + (3.6 x 105) = (1.2x102) + (3600x102) = 3601.2 x 102 Multiplication /Division Multiply/divide numbers by adding/subtracting exponents (1.51 x104) x (2x105) = (1.51x2)x104+5 = 3.02x109 (5.4x105) (2.1x103) = (5.4 2.1)x105-3 = 2.57x102 Moving the decimal point one position to the left adds one to the exponent while moving the decimal one position subtracts one from the exponent. Negative powers of 10 (1.92x10-4) x (8x10-9) = (1.92x8)x10-4+(-9) = 15.3x10-13 = 1.53x10-12 (9.13x10-3) (1.4x10-6) = (9.13/1.4)x10-3-(-6) = 6.5x103 Metric System Base 10 system of measure Primary units of measure are: Mass is gram Length is meter Volume is liter Prefixes indicate size/exponent Kilo = 103 therefore kilogram is 103g Milli = 10-3 therefore milligram is 10-3g (deci, centi) Lab use: Kilo 103 Milli Micro Nano 10-3 10-6 10-9 (10-12) (pico)

Steps are arranged in increments of 103 Scientific International (SI) units are an attempt to standardize reporting of lab values internationally. Units for Basic Properties Length - Meter m Mass - Kilogram kg Time - Second s Current - Ampere A Temp - Kelvin K Amount of substance - Mole M

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Volume is a derived unit that is created by manipulating a basic unit: V = (length)3 = one cubic meter This is a coherent derived unit; that is a unit defined using only the basic unit (length) without any factor. In the laboratory setting, some units are too large to be of practical use; e.g. volume 1 cubic meter = 1 small pickup truck bed Labs use the liter which is a non-coherent derived unit for volume. A liter is 10-3 cubic meters. A liter is 1000 milliliters; a milliliter is a cubic centimeter (10-2 meters). (10-2m)3 x 103 = (10-6)x103 = 10-3 cubic meters Other SI units will be introduced in individual courses. RATIO AND PROPORTION Ratio - relative amounts of one item to a second item. Proportion - relates two ratios; proportion states that two ratios are equal. A ratio is determined by dividing one number into another: Ratio of 5 to 4 can be expressed as 5 4, 5:4, 5/4 It is important to keep in mind what is being compared to what. Example: Test tube containing 1 ml of serum + 9 ml of saline Serum/Saline 1:9 Saline/Serum 9:1 Serum/Tot vol 1:10 Saline/Total Vol 9:10 Tot Vol/Saline 10:9 Tot Vol/serum 10:1

All of the above are examples of ratios created by mixing 1 ml of serum with 9 ml of saline. Ratios and proportion in the lab are used to determine an unknown ratio from a given known ratio. A is to B as C is to D A:B = C:D It is important to set up ratios (known & unknown) in same order & same units.

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If you have 20 grams of a substance in 100ml then how many grams in 20 ml? Known ratio 20g/100ml Unknown ratio Xg/20ml (1) (2) (3) (4) (5) 20g/100ml = Xg/20ml (100ml) x (Xg) = 20gx20ml Xg = (20gx20ml)/100ml Xg = (400g/ml)/100ml X = 4g

Therefore 20g/100ml = 4g/20ml Notice that for all lines in the calculation the units on both sides of the equals sign are the same. In line 2 the units are ml-g on both sides; in lines 3 and 4, ml cancels 1/ml so that g = g. Always check the units on both sides of the equals sign; if they are not the same the equation has been written incorrectly. Factoring & Conversions Temperature 3 scales Kelvin, Celsius/Centigrade, Fahrenheit Kelvin 0K = absolute lowest temperature Celsius 0C = Freezing point pure H2O Fahrenheit 0F = Freezing point salt saturated H2O A change of 1K is equal to a change of 1C. However a change of 1F is not equivalent to a change of 1C or 1K. Range (in degrees) from freezing point to boiling point of H2O: K = 100 C = 100 F = 180 CONVERTING BETWEEN KELVIN AND CELSIUS: Since 1C = 1K and the number of degrees between the freezing and boiling point of water are the same, the two scales must differ by the addition or subtraction of a specific factor. First compare the freezing point of water for the two scales: 273K = Freezing pt. H2O on the Kelvin scale 0C = Freezing pt. H2O on Celsius scale THEREFORE XC + 273 = YK 100C (boiling pt of H2O) = 373K

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CONVERTING BETWEEN FAHRENHEIT AND CELSIUS: Since 1C does not equal 1F it is necessary to adjust for the difference in the size of the degrees. Divide the range of temperature between the freezing and boiling point of water for each scale: 180F/100C = 1F/XC Therefore: 1F = 5/9C or 1C = 9/5F Now adjust for the difference at the freezing point of water: 0C =32F Therefore: F = (Cx9/5)+32 C = (F-32)x5/9 (equate 2 scales and adjust freezing point.) (adjust freezing point and then equate scales)

CONVERTING BETWEEN KELVIN AND FAHRENHEIT: First convert F to C and then C to K UNIT CONVERSION: Ratios and proportion can be used to convert among other units of measure; i.e. English to Metric, Metric to English. With enough time and determination it is possible for the student to derive all the necessary conversion factors in a manner similar to that used to convert among the different temperature scales. The information listed below can be used to develop factors for converting among commonly used units of measure. LENGTH 1 meter (m) = 39.37 inches 2.54 centimeters (cm) = 1 inch 30.48 cm = 1 foot 1.61 kilometers (km) = 1 mile 1 kilogram (kg) = 2.205 pounds 453.6 grams (g) = 1 pound 28.35 g = 1 ounce 1 liter (L) = 1.057 quarts 29.57 milliliters (ml) = 1 fluid ounce 3.78 L = 1 gallon (U.S.)

MASS

Volume

In addition to being used to convert among units of measure, factors can be developed to relate or convert among other types of units or to simplify calculations. Example: Convert amount of nitrogen assayed in a blood test to amount of urea present. This needs to be done since some tests assay for nitrogen not urea. The test assumes that all the nitrogen detected is in the form of urea. Molecular Weight (MW) of nitrogen = 14 Urea consists of two atoms of nitrogen, one carbon four hydrogen. Thus, the molecular weight of urea is 60. Factor: 60 parts urea/28 pts N = X pts urea/1 pt N Parts urea nitrogen x 2.14 = parts urea Example: Determine how much NaCl is needed to yield a given concentration of Cl-?

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MW Na = 23 MW Cl = 35.5 58.5/35.5 = 1.65 = X parts NaCl/1 pt Cl To yield 100 mg Cl, 165 mg NaCl is needed. CONVERTING AMONG METRIC UNITS The use of the base 10 system makes converting among different metric units easy. In general, subtract the exponent of the desired unit from the exponent of the given unit to determine the exponent for the conversion factor. Example: Convert 7.5 milligrams (mg) to nanograms (ng). First convert each unit to the base unit. In this example convert mg and ng to grams. 1 mg = 10-3g 1 ng = 10-9g Subtract exponents: -3 - (-9) = 6 7.5mg = (7.5 x 106) = 7.5x106ng Alternate Method: (10-3g/mg) (10-9g/ng) = (10-3/10-9)ng/mg = 106ng/mg 7.5mg x 106ng/mg = 7.5x106ng

Correction Factor At times it is necessary to perform a test or assay with other than the specified amount of material. In general correction for variation in procedure quantities follows the general rule: Correction Factor = amount specified/amount used Example: Need 5 ml of urine for sodium determination; received only 3.5 ml Take 3 ml and dilute to 5 ml with H2O 5/3 = 1.67 1.67 x Test result = corrected answer Test result 40 mEq Na 40 x 1.67 = 66.8 mEq Na in original specimen

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SOLUTIONS
A solution consists of two components; the dispersed phase (solute) and the dispersing phase (solvent). True solutions contain particles <1nm in size; essentially individual atoms or molecules. Colloidal solutions contain particles 1-200nm in size; exceptions are solutions of large macromolecules such as DNA, RNA protein, lipids. Suspensions contain particles > 200 nm in diameter. Miscible - substances that form solutions, immiscible - substances that don't form solutions. The state varies with conditions of concentration and temperature, etc. Expressions of Concentration There are three different ways to express concentration as a ratio of solute to solvent: weight to weight (W/W), weight to volume (W/V), and volume to volume (V/V). W/W - most accurate but least practical, not affected by changes in T which can effect volume. W/V and V/V - generally accurate under conditions commonly found in the lab setting. Percent Solutions are expressed as parts of solute/100 parts of solution can be W/W, W/V or V/V. MOLARITY Moles of solute/liter solution Mole = is the number of grams equal to the atomic or molecular weight of substance, that is gram molecular weight. MOLALITY Moles of solute/Kg of solvent, this is a W/W expression. Molality is not a practical expression of concentration for routine laboratory use. MOLARITY Moles of solute/liter of solution; a common unit of expression in the lab. NORMALITY Normality is defined as the number of equivalent weights per liter of solution. The Gram Equivalent Weight is defined as the amount of substance that will replace or combine with one mole of hydrogen ions. The gram equivalent weight is calculated by dividing the gram molecular weight by the valence (number of ion states) of the substance. Example: NaCl has a valence of +1 in solution; for every molecule of NaCl, there is one molecule of Na+ that can replace a molecule of hydrogen ions. For H2SO4, there is the equivalent of two molecules of H+ for every molecule of H2SO4. Therefore the gram equivalent weight of H2SO4 is equal to the gram molecular weight (98) divided by 2; 98/2 = 49.

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Calculations involving solutions Making a specific volume of a specified solution. Example: Preparing working solution of the disinfectant Osyl from the stock solution requires diluting the stock one part Osyl to five parts water. This a V/V solution. In this situation you have 250 ml of water so the question is: how much Osyl is needed? 1 pt Osyl/5 pts H20 = X ml Osyl/250 ml 1/5 = x/250 5X = 250 yields X = 250/5 = 50ml of Osyl stock Changing Concentrations To change from one concentration of a solution to another the general formula given below can be used: V1 X C1 = V2 X C2 The units for concentration must be in same. This formula is commonly used when diluting stock solutions. Example: How many milliliters of a 2% solution can be made from 20 ml of 4% sugar solution? 20 ml x 4% = X ml x 2% X = (20 x 4)/2 X = 40 ml Example: What is required to made 100 ml of a 3% solution starting with a 30% stock? 100 ml x 3% = X ml x 30% 300 = 30X 10 = X ml To make the 3% solution dilute 10 ml of the 30% stock to 100 ml using water. Mixing Solutions The final concentration of a solution created by mixing different volumes and different concentrations of the SAME SOLUTE can be determined by using the following formula: (V1 X C1) + (V2 X C2) + (V3 X C3) + ...(Vn X Cn) = Vf X Cf As with the simple formula used above, the units of volume and concentration must be the same. This formula IS NOT USED to determine the concentration of solutions created by mixing two solutions with different solutes together.

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Example: What is the final concentration of a solution created by mixing 20 ml of 5% NaCl with 30 ml of 10% NaCl? (20ml x 5%) + (30ml x 10%) = (20 + 30)ml x Cf 400ml-% = 50ml x Cf Cf = (400/50)% Cf = 8% Working with Percent Solutions W/V solutions are made by mixing solid solute liquid solvent: % = g/100 ml When making a W/V percent solution, the solvent is added to solute until specified volume is reached. This is referred to as "diluting up" or "bringing up" to the specified volume. This procedure is often abbreviated qs for quantity (of solvent) sufficient to yield the specified final volume. This procedure is performed in order to compensate for the volume that the solute occupies in the final solution. Example: Make 100 ml of 10% KOH (potassium hydroxide). It is assumed that the KOH is 100% pure. (Correcting for impure solutes is discussed below) Therefore, the solution is created by adding 10g KOH to a volume of water that is less than 100ml. After the KOH has completely dissolved, water is added bring the final volume to 100ml. Example: How much of a 0.9% NaCl (saline) solution can be made from 2.5 grams of pure NaCl? 0.9% = 0.9g/100ml 0.9g/100ml = 2.5g/Xml Xml = (2.5g x 100ml)/0.9g Xml = 278ml The specified solution is made by adding 2.5 g of NaCl to water and adjusting the final volume to 278ml. For percent solutions using liquid solutes, set up the problem as a ratio and proportion; V1C1 = V2C2. It is important to keep in mind the type of percent solution specified; V/V or W/V. If W/V is specified, the required WEIGHT of solute is used and solvent is added to bring the total volume up to the specified amount. Fortunately most percent solutions made in the lab using liquid solutes are V/V type. Example: Make 50ml of a 5%v/v solution of ethanol (ETOH). The stock solution is absolute (100%) ETOH. 5%v/v = 5ml/100ml Xml/50ml = 5ml/100ml Xml = 2.5ml of absolute ETOH Mix 2.5ml of absolute ETOH with enough water to yield a final volume of 50 ml.

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Molarity A 1 Molar solution is defined as containing 1 mole of a substance per liter of solution. 1M = 1GMW/L (g/mole) x (moles/L) = g/L (g/mole) x (moles/L) x L = g Example: How many grams of H3BO3 (Boric acid, MW=61.83g/M) are required to make 2 liters of a 2M solution? 61.83g/M x 2M/L x 2L = Zg Z = 247.32g Example: What is the molarity of a solution that has 60g of NaOH (sodium hydroxide, MW=40g/M) in a total volume of 2 liters? 40g/M x XM/L x 2L = 60g XM/L = 60g (40g/M x 2L) XM/L =(60/80)M/L X = 0.75M/L = 0.75M Example: How many liters of 1M NaOH can be made with 80g of substance? 40g/M x 1M/L x XL = 80g XL = 80g (40g/M x 1M/L) XL = (80/40)L X = 2L When working with a compound that is less than 100% pure, it is necessary to develop a correction or "purity" factor to correct for the difference between the desired pure substance and the material on hand. This type of problem can be worked either by first assuming 100% purity and then correcting back for the material on hand or by first developing a correction factor. To develop the correction factor convert the per cent assay to the decimal equivalent (purity). grams pure/grams impure = purity grams impure x purity = grams pure grams impure = grams pure/purity Correction Factor = 1/purity g/M x M/L x L x CF = g Example: How much of a solute (MW = 75, % assay = 85%) is needed to make 500ml of a 2M solution? CF = 1/purity = 1/.85 = 1.18 g/M x M/L x L x CF = g 75g/M x 2M/L x 0.5/L x 1.18 =Xg X = 88.5g

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Therefore, to make this solution add 88.5g to enough water to yield 500 ml. Note that if a pure solute was used only 75g would be needed. 75g times the CF (1.18) equals 88.5g. When working with liquids it is necessary to convert the volume to the equivalent number of grams present. This can be done by using information provided in the density of the solution in question. Density is mass per unit volume and is expressed as g/ml. Often the manufacturer lists the specific gravity (SG) instead of the density. Specific gravity is the weight of solid in one ml of liquid divided by the weight of one ml of H20 of 4C. This is a unitless number. However, since 1ml of water weighs 1g at 4C, specific gravity is the equivalent to density and can be interpreted to have the units of g/ml. In general, the best way to solve this type of problem is to first solve for the number of grams of pure substance that is needed and the convert the grams to ml of solute required. Example: How many milliliters of Glacial acetic acid (HC2H3O2, MW=60.05, SG=1.05, % assay = 99) are needed to make 1L of a 2M solution? For this problem, the % assay can be assumed to be 100. 60.05g/M x 2M/L x 1L = Xg X = 120.1g g/density = ml 120.1/1.05 = 114.4ml This answer "makes sense" since each ml of glacial acetic acid weighs slightly more than one gram. Consequently, fewer ml would be needed to yield the required number of grams. Example: Mow many ml of nitric acid (HNO3, MW=63.01, SG=1.42, % assay=70) are needed to make 2L of a 1.5M solution? In this problem it is necessary to correct for the fact that an impure liquid solute is being used. 63.01g/M x 1.5M/L x 2L = Xg X = 189.03g 189g of pure nitric acid are required. Now correct for the concentrated nitric acid solution on hand. Each ml of nitric acid weighs 1.42g Each ml of nitric acid is only 70% pure Each ml of nitric acid solution contains 1.42g/ml x 0.7 = 0.99g of pure nitric acid. 189.03g 0.99g/ml = 190.94ml of concentrated nitric acid required Normality For chemical reactions normality is more useful since it gives the amount of substance per liter that can replace/react with 1M of H+; that is the grams of substance that is equivalent to 1 mole H+. Normality = Gram Equivalent Weight (EqW). The gram equivalent weight is calculated by dividing the gram molecular weight by the number of valence electrons the molecule has. NaCl ionizes to Na+ Cl-, therefore the valence = 1 H2SO4 ionizes to 2H+ SO42-, therefore the valence = 2

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The grams needed to make a specified normal (N) solution is determined using a modification of the formulas used for molar solutions. g/EqW x EqW/L x L = g (N = EqW/L) Correspondingly, the preparation of normal solutions from liquid and/or impure solutes is done using modified equations for molarity calculations. Example: Make 500ml of a 2N NaCl (MW = 58.3). EqW = 58.3/1 = 58.3 58.3g/EqW x 2EqW/L x 0.5L = Xg 58.3 x 2 x 0.55 = 58.3g X = 58.3g This solution is made by adding 58.3g to a volume of water (<500ml) and then adjusting the final volume to 500ml (qs to 500ml). Example: Prepare 1.5L of 2N HCl (Hydrochloric acid, MW = 36.5, %assay = 38, SG = 1.19). First determine the number of grams of pure HCl required: 36.5/Eq x 2 Eq/L x 1.5 L = Xg X = 109.5g Next correct for the use of an impure liquid solute 1.19g/ml of solute X 0.38 = 0.45g HCl/ml of solute 109.5g 0.45g/ml = 243.3ml Again, this answer makes sense because less than half the volume of concentrated HCl is actually HCl. Therefore, more than twice as many ml of solute than grams of pure solute would be required. Example: Prepare 750ml of a 2N solution of H2SO4 (MW = 98.08,% assay = 96, SG = 1.84). First determine the number of grams of pure sulfuric acid required: EqW = 98.08/2 = 49.04 49.04g/EqW X 2 EqW/L X 0.75L = Xg X = 73.56g Next correct for the use of an impure liquid solute: 1.84g/ml X 0.96 = 1.77g H2SO4/ml of solute 73.56g 1.77 g/ml = 41.56 ml Percent solutions using % assay and specific gravity When preparing percent solutions from impure liquid solutes it is important to determine the type solution desired; V/V, W/V or W/W. It is usually easier to determine the number of grams or milliliters of pure solute required and then to adjust for the % assay.

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Example: Prepare a 20%w/v solution of HCl (MW = 36.5, %assay = 38, SG = 1.19). 20%w/v = 20g/100ml For HCl, 1ml = 1.19g/ml x 0.38 = 0.45g/ml 20g 0.45g/ml = 44.4ml Mix 44.4ml of HCl with enough water to yield a total volume of 100ml. Note that it was not necessary to use the information regarding molecular weight in the solution of this problem. Working with hydrated solutes: Solutes may be obtained both in anhydrous form and in hydrated form. The hydrated form of solute is a compound in which the solute is complexed with the specified number of water molecules. The hydrate is an "impure" form of solute in that the compound consists of a specified amount of solute "diluted" with a specified amount of water. The percent assay or purity can be calculated by dividing the molecular weight of the anhydrous form by the formula weight of the hydrated form. Usually the formula weight is stated on the container. The anhydrous molecular weight can either be looked up in a text or calculated by subtracting the weight of the water from the formula weight of the hydrated form. Example: The formula weight of CuSO45H2O is 249.7. What is the molecular weight of the anhydrous form of copper sulfate and what is the %assay of the hydrate? MW of water = 18.02 5H2O = 5 x 18.02 = 90.1 249.7 - 90.1 = 159.6 159.6/249.7 X 100 = 63.9% assay This information can then be used to make solutions using hydrated forms of the solute. Example: Make 2.75 liters of 3.2M copper sulfate using CuSO45H2O. First determine how much anhydrous copper sulfate is necessary. 159.6g/M x 3.2M/L x 2.75L = Xg X =1404.5g Determine the amount of hydrate needed: 1404.5g/0.639 =2198g of hydrate is required. This same overall approach can be used to determine normality, percent solutions, etc. Conversion of Concentration Expressions CONVERTING BETWEEN MOLARITY AND NORMALITY Remember that normality is the number of gram equivalent weights per liter and gram equivalent weight is equal to the gram molecular weight divided by the valence of the solute. N = M x Valance M = N/Valance

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CONVERTING MOLARITY TO PERCENT SOLUTION MW x M = g/L Therefore M = (g/L)/MW M = (g/L)/MW = (g/100ml x 10)/MW M= (%w/v x 10)/MW Example: Convert a 30%w/v solution of NaCl (MW = 58.5) to a molar solution. M = (% x 10)/MW M = (30 x 10)/58.5 M = 5.13 Therefore 30% solution of NaCl is a 5.13M solution. Example: Convert a 6M solution of NaOH (MW = 40g) to a %w/v solution. Rearrange the formula from above: M x MW = (% x 10) (M x MW)/10 = % 6x40/10 = 24% A 6M solution of NaOH is equivalent to a 24%w/v solution. CONVERTING A NORMAL SOLUTION TO PERCENT SOLUTION g/EqW x N = g/L N = (g/L) EqW N = (% x 10) EqW Example: Convert a 3N H2S04 (MW = 98, % assay = 96, SG = 1.84) solution to a %w/v solution. N = (% x 10)/EqW % = N x EqW/10 % = 3x49/10 % = 14.7 %w/v Note that for this problem the information on % assay and SG was not needed since the original normality concentration accounted for this. CONVERSION OF mg/dl TO mEq/L The old unit for reporting out the amount of sodium, potassium or chloride ions in body fluids was mg/dl. The new standard for reporting is mEq/L (milliequivalents/L or "millinormals"/L). Depending on the procedure used to measure these ions in solution, it may be necessary to convert from the old to the new units. It may also be helpful to some physicians to have the values converted to the older, more familiar units of concentration. mEq/L = mgEqW/L (milligram equivalent weight/L) mEq/L = (mg/L) EqW = (mg/dl x 10)/EqW.

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Example: Convert 300mg/dl of Cl- to mEq/L of Cl-. mEq/L = (300 x 10)/35.5 = 3000/35.5 mEq/L = 84.5 Therefore 300mg/dl of Cl- is equivalent to 84.5mEq/L. Example: Convert 150 mEq/L NaCl to mg/dl of NaCl. mEq/L = (mg/dl x 10)/EqW mg/dl = (mEq/L x Eq)/10 mg/dl = (150 x 58)/10 870 mg/dl NaCl CONVERSION OF COMPLEX METRIC UNITS Conversion among complex metric units of concentration is an extension of conversion among metric units of measure. The complex unit of concentration, for example mg/dl is broken into its component parts, mg and dl. Each of these is then converted to the corresponding desired unit of measure, for example and ml. g Convert mg/dl to g/ml = 10-3mg g mg x 103 = g ml = 10-2dl dl x 102 = ml mg/dl x 103/102 = mg/dl x 101 = g/ml Example: Convert 21g/L to mg/ l. g x 103 = mg L x 106 = l 21 x 103/106 = 21 x 10-3 21g/L = 2.1 x 10-2 = 0.021mg/ l Example: Convert 316 g/ml to mg/dl.

x 10-3 = mg g ml x 10-2 = dl 316 g/ml x 10-3/10-2 = 316 x 10 -1 = 31.6mg/dl

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DILUTIONS
General Concepts Many laboratory procedures, especially those in Immunology/Serology, require that a series of dilutions be performed on serum or other body fluids. These dilutions are used to establish a relative measure of the quantity of antibody or antigen present in the fluid. This section will review the steps necessary to make simple and serial dilutions. A dilution is expressed as the parts of solute relative to parts total volume. This is written using a slash (/). The colon (:) is used to express a ratio; the parts of one substance relative to parts of a second substance. Therefore, dilution is an expression of relative concentration. The dilution of a substance can be determined by dividing the volume of the solute by the volume of the solute + solvent or the total volume. Example: What is the dilution when 1ml of serum is mixed with 9ml of saline? dilution = 1ml serum (1ml serum+ 9ml saline) dilution =1/10 Therefore the serum has been diluted 1 to 10 with saline. Example: What is the dilution achieved when 3ml of serum are mixed with 22ml of saline? dilution = 3ml serum (3ml of serum + 22ml saline) dilution = 3/25 Express this dilution as 1 to X dilution. 3/25 = 1/X X = 25/3 = 8.33 Therefore 3/25 is the same as 1/8.33. The volume of solute and solvent required to make a given volume of a specific dilution can be determined using ratio and proportion. The specified dilution represents one ratio and the amount of solute required becomes the unknown in the second ratio. Example: How many milliliters of serum are required to make 300ml of a 1/10 dilution? 1 part serum/10 parts total volume = Xml serum/300ml total volume 1/10 = X/300 X = 300/10ml X = 30ml This solution is made by diluting 30ml of serum up to 300ml with saline. The amount of undiluted solute present in a specified volume of a given dilution can also be calculated using ratio and proportion.

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Example: How many milliliters of serum are present in 25ml of a 1/8 dilution. 1 part serum/8 parts tot. vol. = Xml serum/25ml tot. vol. 1/8 = X/25 X = 25/8 = 3.125ml Therefore there are approximately 3.1ml of undiluted serum in 25ml of a 1/8 dilution. In the preceding examples, the concentration of serum is implied to be 100%, that is a 1/1 dilution. When diluting solutions with a specified concentration the final concentration can be calculated by multiplying the original concentration by the dilution. Example: What is the final concentration of a solution created by diluting 5ml of 5N NaCl to a final volume of 500ml? First calculate the dilution: 5/500 = 1/100 Multiply the original concentration by the dilution: 5N X 1/100 = 0.05N Therefore the final concentration of the solution is 0.05N (5x10-2N) Dilution Series In some laboratory situations it becomes necessary to perform a series of dilutions on a given solution. This usually done for two reasons. First, the desired dilution may be technically impractical. For example it is difficult to make a 1/10,000 dilution in a single step. This would require accurately diluting 1ml of starting solution to a final volume of 10,000ml. In this situation, the volume of the solute used is smaller that the inherent inaccuracy of the measuring device for the solvent (the error is usually + 1%; in this case + 10ml). Therefore it becomes necessary to perform a series of small dilution steps to achieve the final desired concentration. Second, when determining the relative concentration of antibody or antigen in a patient's serum, the usual procedure is to dilute the serum sample repeatedly in a series of steps with the same dilution factor. That is, repeatedly dilute the serum 1/10 or some other dilution. When performing this type of dilution procedure, it is necessary to keep track of two dilutions: the tube dilution-the dilution factor of that particular tube or container; the solution dilution-the actual dilution of the starting substance in that particular tube. In both situations the tube dilution is calculated by dividing the #parts solute by #parts total volume. The solution dilution of a given tube is determined by multiplying the tube dilution of the specified tube by the solution dilution of the preceding tube. Notice that the tube dilution is independent of the dilution of the preceding tube while the solution dilution depends on the dilutions performed in the preceding tube(s). NOTE! When serum is diluted, undiluted serum is assumed to be 1/1.

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Example: One ml of serum is added to 4ml of saline (tube#1). One ml of this dilution is added to 9ml of saline (tube#2). One ml of this dilution is added to 99ml of saline (tube#3). Determine the tube and solution dilution in each tube. Tube # Tube dilution Solution dilution 1 1/(1=4) = 1/5 1/1 X 1/5 = 1/5 2 1/(1+9) = 1/10 1/5 X 1/10 = 1/50 3 1/(1+99) = 1/100 1/50 X 1/100 = 1/5000

Therefore the final dilution of serum in Tube#3 is 1/5000. Example: What would be the concentration of HCl present in Tubes#1-3 if 10N HCl was diluted in the manner described above? For each tube, the concentration of HCl is determined by multiplying the initial concentration of HCl by the solution dilution for that tube. Tube#1 = 1/5X10N = 2N HCl Tube#2 = 1/50X10N = 0.2N HCl Tube#3 = 1/5000X10N = 10 5X103 = 2X10-3N HCl Note that the final expression of concentration is in the same units as the initial concentration; in this example normality. If the initial concentration is given in percent then the final concentration is given in percent. For serum the concentration is reported as the dilution since the concentration of undiluted serum is 1/1. Preparing dilutions of specified volume It is often necessary to produce a specified volume of a dilution or to produce a series of dilutions of specified volume from a limited amount of starting material. It then becomes necessary to determine how the make each dilution. Example: Prepare a series of dilutions of serum (1/10, 1/100, 1/500) so that the total volume of the final container is 100ml and starting with only 1.5ml of serum. The easiest way to solve this problem is to establish a series of dilutions where and only 1ml of serum is used for the first dilution and the final tube has a volume of 100ml. Tube#1 = 1/10=>1ml of serum + 9ml of saline Tube#2 = 1/100 = 1/10 x X => X = 1/10 Tube#3 = 1/500 = 1/100 x Y => Y = 1/5 1/5 = Z/100 Z = 20ml To prepare this dilution, first dilute 1ml of serum 1/10. Take 2ml of Tube#1 and dilute it 1/10. To prepare 100ml of the final dilution take 20ml of tube#2 and dilute 1/5 to yield a final volume of 100ml. Remember the following general rule: final conc. = init. conc. x dil tube#1 x dil tube#2 x ...

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Serial Dilutions Serial dilutions represent a specialized application of dilution procedures. Serial dilutions are performed when it is necessary to dilute a substance in a series of equal dilutions. This is most commonly done when trying to determine the relative concentration of antibody or antigen in a serum specimen. serum samples are generally diluted twofold; a series of dilutions. The same concepts used above are employed when making serial dilutions. Again, the dilution of serum is set as 1/1. Consequently the dilution factor increases by powers of two. Example: A serum specimen is diluted in the following manner: 1ml of undiluted serum is mixed with 1ml of saline (tube#1); 1ml of this tube is added to a second tube containing 1ml of saline (tube#2); this same procedure is repeated twice more (tubes#3-4). What is the tube dilution and solution dilution for each tube? Tube # Tube dilution 1 1/(1+1) = 1/2 2 1/(1+1) = 1/2 3 1/(1+1) = 1/2 4 1/(1+1) = 1/2

Solution dilution 1/1 X 1/2 = 1/2 1/2 x 1/2 = 1/4 1/4 X 1/2 = 1/8 1/8 X 1/2 = 1/16 Note the pattern developing. The solution dilution is equal to raised to the power corresponding to the tube number: tube#1=(1/2)1=1/2 tube#2=(1/2)2=1/4 tube#3=(1/2)3=1/8 tube#4=(1/2)4=1/16 This holds for the original serum if it is labelled tube#0: (1/2)0=1=1/1 Therefore in any serial twofold (2X) dilution series, the tube dilution is always 1/2 and the solution dilution is (1/2)tube#. In many commercial serology kits, the technologist is required to prepare a series of twofold dilutions of a specific volume. It only necessary to determine the volumes required for one twofold dilution and then to repeat that the required number of times. Example: Prepare a series of twofold dilutions of patient serum such that the final volume in each tube is 0.5ml and the final solution dilution is 1/1024. First determine the volumes required for the twofold dilution: 1/2 = x/0.5ml => X =0.25ml However, if this result is used to produce the series, the FINAL VOLUME in each tube will be 0.25ml since 0.25ml must be take from the preceding tube to make the succeeding dilution. Therefore, to insure that the final volume is 0.5ml, double the volumes required for a single twofold dilution. Thus 0.5ml of serum would be mixed with an equal volume of saline. One half of this tube (0.5ml) would be used to make the dilution in the next tube. To determine the number of tubes required to make the series calculate the power that must be raised to in order to yield 1/1024. It helps to have a calculator that can perform exponential calculations. Two raised to the tenth power (210) is 1024; ()10=1/1024. IN SUMMARY: The dilution series requested is prepared by adding 0.5ml of saline to each of 10 tubes. 0.5ml of undiluted serum is added to tube#1 and mixed well. 0.5ml of tube#1 is then added to tube#2. This is repeated out to tube#10. 0.5ml of sample is removed from tube#10 and discarded to yield a final volume in tube#10 of 0.5ml.

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