Errors and Significant figures

Systematic (determinate) -consistent error. Can be detected & corrected Errors Random (indeterminate)-due to limitations of physical measurement. Cannot be eliminated. Systematic error (i) operational & personal e.g. mechanical loss of material during analysis.or inability to detect colour changes at end-point of titration. (ii) instrumental or reagent errors- faulty calibration of balances, attack of reagents upon glassware. (iii) errors of method - incorrect sampling & from incompletenes of a rx. (iv) additive & proportional errors.

Systematic error can be reduced by (a) calibration of apparatus & application of correction factors (b) running a blank determination separate determination with a blank under identical experimental conditions (c) running a control - with a quantity of a standard substance with approx the same weight of constituent as contained in sample. (d) use of independent method of analysis (e) running parallel determinations - serves as a check on the precision of result-does not guarantee accuracy (f) use of standard addiion/internal standard or isotopic dilution. (g) amplification methods - useful when small amount of material is to be analysed.

CH4304 Laboratories

Significant figures and computations. 1. The number of significant figures in a value is the minimum number of digits required to write the value in scientific notation. e.g. 0.00035 = 3.5 x 10-4 i.e. two significant figures. 0.000 0045 = 4.5 x 10-6, two significant figures. 2. The first uncertain digit in a calculated result should be the last significant figure. e.g.

0. 002364 0. 000003 =0 . 0946 0 .0002 4 0. 02500 0. 00005 2. 364 x10-3 2. 500 x10 -2 righ

3. For addition and subtraction the last significant figure is determined by the decimal place of the least certain number 21.29965 + 12.2 = 33.5

4. For multiplication and division the number of figures is usually limited by the factor with the least number of digits. Retain in each factor one more significant figure than is contained in the factor having the largest uncertainty e.g. 1.26 x 1.236 x 0.6834 x 24.8652 should be carried out as 1.26 x 1.236 x 0.688 x 24.87 and the result expressed to three significant figures

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5. For values expressed as logs the number of figures in the mantissa should
equal the number of significant figures in the original value.

Calculating Uncertainty

1. For addition and subtraction use absolute uncertainty

2 2 e 4= e 2 1 e 2 e 3

e.g.

1.76 ( 0.03) + 1.89 ( 0.09) - 0.59 ( 0.02) = 3.06 e4 = 3.06 0.041

1. For multiplication and division use relative

e = e 2 1 .76 0 . 03 x1 .89 0 . 02 0 . 59 0. 02

uncertainties

relative uncertainty (0.03)/(1.76) x 100 = 1.70%

e = 1 . 7 0 2 1 . 0 6 2 3 . 3 9 2 = 3 . 9 4 5 .6 4 3. 9 4 = 5 .6 0. 2

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Mean & Standard Deviation

When a quantity is measured with the greatest exactness of which the instrument, method and observer are capable of it is found that the results of successive determinations differ among themselves to a greater or lesser degree the average value is accepted as the most probable this may not always be he true value Difference between the measured and the true value = absolute error and this is a measure of the accuracy of a result. The true or absolute value of a quantity cannot always be established experimentally so the observed value must be compared to the most probable value Sample mean

x= i

xi
n

The spread of values is measured most efficiently by the sample standard deviation s

s= i

x(-)i2
n 1

s2 = variance R.S.D. relative standard deviation

= s x

Sometimes referred to as the coefficient of variation C.V.

s x 1 0 0 C= . V . x
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Example 1 Analysis of a sample of iron ore gave the following % values for the iron content: 7.08 7.21 7.09 7.16 7.14 7.07 7.14 x = 7.13 s = 0.047 C.V. = 0.66% 7.18 7.11

The greater the number of measurements (n) the closer the sample mean (x) approach the population mean (). The standard error of the mean sx
s x s = n

For example 1 sx = 0.014. If 100 measurements were made denominator =100 1/2 so sx = 0.0047 so the precision of a measurement may be improved by increasing the number of measurements.

Distribution of standard errors If a large number of replicate observations are made (>50) of a continuous variable the results obtained will usually be distributed about the mean in a roughly symmetrical manner-Gaussian Error Curve.

12 y= (-ex) /2 2

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68% of all values fall within one S.D. of the mean, 95% will fall within 2 S.D (1.96). and 99.7 will fall within 3 S.D of mean. From e.g. 1 S.D. = 0.047 so 68% of values will lie 0.047 of the mean. 95% will lie between of the mean value i.e. there is a 5% (1 in 20) os a resultdiffering from the mean by more than 0.094%.

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Reliability of results
Statistical figures obtained from a set of data are of limited use by themselves a) reliability of results Analysis of results b) comparison of results with true value or with other sets of data

Rejection of results should be considered only when a suitable statistical test has been applied or when there is an obvious chemical or instrumental reason E.g. 2. The following data were obtained for the determination of Zinc in a sample of dust: 4.3 g g-1, 4.1g g-1 , 4.0g g-1 , 3.2g g-1 . Should the value 3.2 be rejected. Apply Q test questionable value - nearest value largest value - smallest value 3.2 - 4.0 4.3 3.2 0.8 = 0.727 1.1

Q=

If Q (observed ).>Q (tabulated) discard the questionable point. Q for 4 points from table = 0.831 so result should be retained.

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Correlation and regression calibration: construct calibration curves by measuring analytical signal for each standard & plotting against concentration two statistical operations should be performed on calibration curve

Correlation coefficient {r}: to determine if graph is linear or

in the shape of a curve

Least square error associated with best fit line/curve

nxi y i - x i y i
2 2 2 [ nx 2 i xi ][ ny i y i ]

r=

n: number of data points

r must be close to 1 for good fit

Linear Regression- least squares 2 assumptions

errors in y values substantially greater than errors in x values uncertainties (standard deviations) in all y values are similar
eqn of line y = mx + b

point (xi, yi) vertical deviation (di) = yi - y di = yi-y = y-(mx + b) di2 = (yi - mx - b)2 : least squares

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 xi yi xi n yi
rli D b=
2

xi xi yi xi yi
rli D D=
2

xi x i n xi
rli m= Standardised Acid and Standardised Base CH4304 Laboratories 9

Standard HCl Procedure 1. of concentrat (37 wt %) HCl that should be added to 1 L of distilled water to produce 0.1 M HCl, and prepare this solution. Use the information provided in Table 1 to calculate the volume

2.

Dry primary grade sodium carbonate for 1 hr at 110oC and cool it in a desiccator. Weigh four samples containing enough Na2CO3 to react with 25 mL of 0.1M HCl and place each in a 125 mL flask. As you are ready to titrate each one, dissolve it in about 25 mL of distilled water. 2HCl + Na2CO3 CO2 + 2NaCl

3.

FW = 105.989 Add 3 drops of bromocresol green indicator to each and titrate one rapidly (to a green colour) to find the approximate end-point. 4. Carefully titrate each sample until it just turns from blue to green. Then boil the solution to expel CO2. The solution should return to a blue colour.

5. Carefully add HCl from the buret until the solution turns green again.
A blank titration can be performed by using 3 drops of indicator in 50 mL of 0.05 M NaCl. Subtract the volume of HCl needed for the blank solution from that required to titrate Na2CO3.

6. Calculate the mean HCl molarity, the standard deviation and the
relative standard deviation.

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Standard NaOH

1.

A 50 wt % aqueous NaOH is provided. Normally this solution is prepared in advance and allowed to settle. Na2CO3 is insoluble in this solution and precipitates. The solution should be stored in a tightly sealed polyethylene bottle and handled gently to avoid stirring the precipitate when the supernate is taken. The density is close to 1.50g of solution per mL. Primary standard-grade potassium hydrogen phthalate should be dried for 1Hr at 110oC and stored in a dessicator. COOK
+

2.

COOK NaOH COONa

+

H2O

COOH F.W. = 204.22

3. Boil 1.0L of water for 1Hr to expel CO2. Pour the water into a
polyethylene bottle which should be tightly capped. Calculate the volume of 50% aqueousNaOH needed to produce 1.0L of approximately 0.1 M NaOH. transfer this to the water bottle, mix well and allow to cool.

4.

Weigh four samples of the solid KH(C8H4O4) (MW 204.23)and dissolve each in approx. 25 ml of distilled water in a 125ml flask. Each sample should contain enough solid to react with about 25ml of 0.1M NaOH. Add 3 drops of phenolphthalein ndicator to each and titrate one of them rapidly to find the approximate end-point. The burette should be fitted with a loose fitting cap to minimise entry of CO2. Calculate the volume of NaOH required for each of the other three samples and titrate them carefully swirling the flask occasionally. The end point is the firstappearance of a faint pink colour. This colour will disappear as CO2 from the air dissolves in the solution. Calculate the average molarity, the standard deviation, and relative standard deviation.

5.

6.

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Name

Approximat e weight percent

Approximat e molarity

mL of Reagent needed to prepare 1 L of 1.0 M solution 57.5 82.6 62.9 85.5 67.6 55.6 69.0 51.5 85.5

Acetic acid Hydrochloric acid Nitric acid Perchloric acid Phosphoric acid Sulphuric acid Ammonia* Sodium Hydroxide Potassium hydroxide

99.8 37.2 70.4 70.5 85.5 96.0 28.0 50.5 45.0

17.4 12.1 15.9 11.7 14.8 18.0 14.5 19.4 11.7

*28wt% ammonia is the same as 56.6 wt% ammonium hydroxide

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Preparation of buffers
Buffer solution can be defined as an equilibrium mixture of a conjugate acidbase pair which is capable of resisting substantial; changes in pH upon the addition of small amounts of acidic or basic substances. With some exceptions, either the acid or the base of the conjugate pair is a weak electrolyte. The Henderson-Hasselbalch equation is a mathematical statement which defines the pH of a solution of a conjugate acid-base pair in terms of the dissociation constant of the weak acid-base pairs and the equilibrium concentrations of the acid and its conjugate base. This equation is extremely useful in calculating the pH of the buffer. Figure 1: Preparation of a 0.02M Tris buffer at pH 8.0.

Buffer A and B are prepared in the same way except 0.5 mole of NaCl is added to flask B before the volume is brought to 1 L for buffer B.

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Ion-exchange chromatography - spectrophotometric determination of the composition of a sample of VOSO4

VOSO4 supplied commercially contains impurities H2SO4 and H2O. A solution will be prepared from a known mass of reagent. The VO2+ can be assayed spectrophotometrically and the total cation (VO2+ and H+) can be assayed by ion-exchange. Together these measurements enable the determination of the quantities of impurity in the sample. Ion exchange resins are polymer matrices containing charged site which are used to separate charged species . CH=CH2 CH=CH2

Styrene

CH=CH2 divinylbenzene

CH-CH2

CH-CH2

CH-CH2

CH-CH2

SO3-H+ CH-CH2

CH-CH2

SO 3-H+ CH-CH2 CH-CH2

SO 3-H+

SO3-H+

Anion exchangers contain bound positive groups e.g. NR3+ Cation exchangers contain bound negative groups e.g. SO3-

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Procedure It is initially necessary to determine the ion-exchange capacity of the resin (moles of Na+/gram of resin). Weigh accurately about 2.000g of Amberlite 120A resin in the Na form. Make a slurry of the resin in distilled water and pour into the glass chromatography column. The level of the water should always cover the resin. Never let the resin run dry
NaCl

HCl

Na
+

+ +

H H

+ +

Na Na

Na H H
+ +

+ +

Na Na

Na

Na

titrate with NaOH NaCl HCl

VOSO4.H2SO4

VO2+ H
+

VO 2+

H 2SO 4

titrate with NaOH

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2. Conversion of the resin to the H+ form. Pipette 5.0mL of 1.0M acid (H2SO4 or HCl) onto the column and allow to equilibrate. Wash off excess acid with the distilled water until a neutral eluate is obtained (use litmus paper). 3. Determine ion exchange capacity of resin in moles Na+/g resin. Pipetted 2.0mL of standardised 0.500M NaCl solution onto the column and allow to equilibrate for 5 minutes. Wash with about 25 mL of distilled water and collect the eluate in a 150mL flask. Add three drops of phenolphthalein indicator and titrate with standard 0.100N NaOH.

4. VOSO4 solution: weigh accurately, approximately 0.50g of VOSO4 and

make up to 100mL with distilled water. Using the molar absorptivity ( ) of the vanadyl ion at 750nm, VO2+ = 18M-1cm-1 it is possible to calculate the amount of pure VOSO4 in the sample.

5. Following regeneration of the resin as previously described, (2 above) the

procedure is repeated for the vanadyl sulphate solution to determine the total cation content, i.e. VOSO4 + H2SO4 .Care should be taken not to exceed the exchange capacity of the resin.

4. The water in the sample can be calculated from a accurate measurement of the initial weight. i.e. original weight - (VOSO4 + H2SO4) 5. Express your answer as (VOSO4)x(H2SO4)y(H2O)z Example

(a) In an experiment to determine the purity of Vanadyl Sulphate

2.000 g of ion-exchange resin in the H+ form were used. 20.00 ml of 0.100M NaCl was put on the column and allowed to equilibrate. The eluate was collected and titrated against 0.010M NaOH, the titre was 32.0 ml . What is the ion-exchange capacity in mols/g of resin.

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Ans 32.0 x 0.010 M = 3.2 x 10-4 mols NaOH = 1.6x10-4 mols /g resin

(b)

b)

Assuming the VOSO4 was absolutely pure what is the

maximum volume of solution that can be put on the column such that the ion-exchange capacity is not exceeded given that a solution of Vanadyl sulphate was prepared by dissolving 0.1420g in 100mls of H2O. . Ans: VOSO4 mol wt = 163 g/mol so if 100% pure the 0.1420g of VOSO4 = 8.7 x 10-4 mol. Resin capacity for VO2+ ion is 1.6x10-4 mols, so the maximum capacity of the resin for VOSO4 (if 100% pure ) is 18.37 ml of solution

(c) c) A solution of Vanadyl sulphate was prepared by dissolving

0.1420g in 100mls of H2O. 10.0 ml of the solution was put on the regenerated resin (H+ form) and allowed to equilibrate. The eluate was collected and titrated against 0.010M NaOH. The titre was 18.0 ml. The absorbance of the solution in a 1cm cuvette was 0.126 at 750nm when =18cm-1M-1. (VOSO4)1.00 (H2SO4)y(H2O)z Ans: A = c so c= A/ = 0.126/18 = 7.0 x10-3M in 10 mL 7.0 x10-5 mol VOSO4 = 0.01141 g Titre 18.0 ml 18 .0 0 .010 M mol of (VOSO4 + H2SO4) 1000 2
.

Calculate the molar ratio as

= 9 x 10-5 mol 2.0 x 10-5 mol H2SO4 = 1.96 x10-3 g H2SO4 wt of H2O = 0.01420 (in 10 mls) (0.01141 + 1.96 x10-3)g = 8.3 x 10-4 g H2O = 4.6 x10-5 mol (VOSO4) 7.0 x 10-5 (H2SO4) 2.0 x 10-5 (H2O) 4.6 x10-5

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(d)

= (VOSO4)1.00 (H2SO4)0.29(H2O)0.66

Gas Chromatography
Gas chromatography is an instrumental technique for the separation of volatile molecules. The mobile phase is an inert gas generally nitrogen or helium and the stationary phase is a nonvolatile liquid chemically bonded onto a quartz capillary column of very narrow diameter.. The column is placed in a temperature programmable oven and the Nitrogen passes through it. Gas chromatography involves a sample being vapourised and injected onto the head of the chromatographic column. Samples are injected onto the column via a heated injection port and are transported through the column by the flow of inert, gaseous mobile phase; and are separated on the basis of having different affinities for the stationary phase. As each component exits (or elutes) from the column it enters a detector and produces a signal peak. The peak area is proportional to the concentration or mass of that component in the sample. Qualitatively, the identity of unknown components can be determined by comparing their retention times with known standards. Quantitatively, the concentration of a given component can be determined by measuring the peak area. The samples are detected as they exit the column. The integrator gives an output of peak height versus time, called a chromatogram from which samples may be identified and concentrations evaluated.

injector detector
N2

integrator

column

oven

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Background Chromatographic techniques are used to perform qualitative and quantitative analysis on complex mixtures. As the mixture of components to be separated is carried through thecolumn, separation is affected because of the varying attraction each component of the mixture has for the stationary phase. Substancesthat are more strongly attracted to the stationary phase will becarried down the column at a slower rate than those with a lesser attraction.. The temperature of thecolumn and, to a lesser degree, the flow rate of the mobile phase alsoaffect the degree of separation. The time required for a particularcomponent of a mixture to pass from the injection port, through the column, to the detector is its retention time. Under identicalconditions, the same substance will have the same retention time inrepeated analyses. For this reason, retention time can be usedqualitatively to identify a substance in a mixture. If a quantitative determination is desired, the method of internal standards is normally employed. An internal standard is needed when conditions such as sample size of gas flow cannot be replicated exactly between trials. A compound is chosen to serve as an internal standard that has physical properties similar to those of the substance you wish to measure. A calibration curve is constructed in which the ratio of the peak area of the compound of interest (the analyte) to that of the internal standard is plotted on the y-axis versus the concentration of the analyte on the x-axis. The peak area ratios will be proportional to the analyte concentration, whereas the peak area of the analyte alone will not be. Peak areas are calculated by the electronic integrator.

In this experiment you are provided with an unknown alcohol in water and you are required to 1. identify the unknown on the basis of its retention time using the standards provided

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2. quantify the concentration of butanol in water by internal standardisation using one of the standards provided. 3. calculate N (the number of theoretical plates required to elute the unknown) and the resolution wrt the internal standard
EXPERIMENTAL PROCEDURE: A: Identification of unknown in alcohol mixture

B: quantitative analysis of butanol in a butanol:water mixture. A: identification of unknown. In a 10mL volumetric flask make up 3 standards containing 0.20mL each of butanol, ethanol, and propanol in distilled water. Inject each standard onto the GC and record the retention times. Inject the unknown provided. Comparison of the Rt of each of standards against the components of the unknown should allow direct identification of the unknown(s) The Quantitation of Butanol Prepare a series of standards ranging from 2.0% to 8.0% of butanol as indicated below using pipettes to deliver each liquid. Use the 50ml flasks and make up to the mark with water After the four standards have been prepared, add 2.0mL of propanol to each mixture via pipette. Propanol will serve as the internal standard in this analysis. standard volume of butanol CH3CH2CH2CH2OH 1.0mL 2.0mL 3.0 mL 4.0 mL volume of propanol CH3CH2CH2OH 2.0 2.0 2.0 2.0 mL mL mL mL Total volume

50.0 50.0 50.0 50.0

Swirl each mixture thoroughly to ensure complete mixing, then cover each solution to avoid evaporation.

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Obtain 50.0 mL of an unknown butanol:Water mixture from your instructor. Using a 1.0 mL pipet, add exactly 2.0 mL propanol to the unknown. Swirl the mixture thoroughly and cover to retard evaporation. . SETTING UP THE Gas Chromatograph You will use the HP5890gas chromatograph in conjunction with the integrator The integrator output will list the peak number Peak # 1 2 3 Retention time (mins) x.xx Area counts xxxxxx % Area xx.x

Experiment should be run isothermally. Your instructor will select the temperature

CALCULATIONS: 1. Calculate the ratio of peak area values of butanol to propanol. 2. Using EXCE L prepare a calibration curve by plotting the ratios of concentration of butanol/propanol on the x-axis versus the average of the ratios of the peak area of butanol to peak area of propanol. 3. Use EXCELto perform a regression analysis on the calibration curve . 4. Calculate the concentration of butanol in the unknown sample from the regression analysis basing your calculation on the straight line equation form, y = m x + b. 5. Turn in the graph of the calibration curve, the regression analysis, the calculation of the unknown and the Report Sheet.

A major source of precision error in GC is from poor injection technique. Both manual and automatic injection methods are available. If automatic equipment is available use it.

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If not several approaches can be used to try and reduce error. The first step is to ensure that the syringe is properly filled containing no air bubbles. The instructors will demonstrate how air bubbles can be eliminated .The sample should be injected rapidly as a plug to avoid band broadening and poor resolution. a) draw the sample into the syringe barrel b) draw 2-3l of air into the barrel c) insert needle into injection port and allow to heat for a few seconds d) Inject quickly but without excessive force as the syringes are fragile e) When the syringe has been removed check for any un-injected sample and subtract from nominal injection volume f) Clean the syringe using diluent (water in this case) six times before the next injection. g)

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High Performance Liquid Chromatography

HPLC is a technique suitable for separation of most organic compounds and is the most often encountered instrumental quality control method. Separation occurs between a liquid mobile phase consisting of degassed very high purity organic/aqueous solvents which may or may not be buffered and a stationary phase which is chemically bonded to a support material. The most frequently used stationary phase is Octadecylsilane ODS which consists of a long hydrocarbon chain, it is a non-polar material.

pump

injection loop column

uv-vis detector

mobile phase

integrator

The purpose of this experiment is to determine the concentration of caffeine in common beverages using a reversed-phase column, Equipment and Materials 1. HPLC System with variable wavelength detector. 2. Reversed-phase column (C-18); Ultrasphere ODS, 4.6 x 150 mm, 5 micron particles coated with octadecylsilane groups. 3. 25-L flat nose syringe. B. Solutions the mobile phase is is 40:60 CH3OH:H2O

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Part A:
.

Identify caffeine in beverages on the basis of its retention time (tr)and determine its concentration by standard curve. Stabilize the system with the methanol/water solvent at a flow rate of about 1.0 mL/min. Flush the injection port with at least 3-4 syringe volumes of
mobile phase.

(i) Using the pure caffeine provided make up a stock solution of approx 0.1% (ii) Prepare a series of standards by serial dilution of 0.01%, 0.005%, 0.001%, 0.0005% caffeine in water. (iii) Load 50 L of each onto the column, When the injection loop (20 L) is properly flushed and filled, injected onto column.. Begin recording the chromatograms at an absorbance range (AUFS) such that the entire peak appears on the chromatogram. (iv) note the retention time and the area associated with each peak. Construct a calibration graph, and find the best fit regression line (v) Take a sample of beverage. Filter to remove any undissolved material and inject onto the chromatograph. The caffeine can be identified by its retention time and the concentration calculated from its peak area.
..

Part B You are required to calculate certain performance characteristics of your separation. Prepare solutions of 0.1% NaNO3 , 0.1% benzoic acid and 0.1% caffeine as well a mixture of 0.1% caffeine and 0.1% phenol. NaNO3 will be used to estimate tm as it will not be retained by the stationary phase as it is ionic and will elute with the mobile phase. (vii) Inject 50 L of each onto the column, note the retention time. The chromatograms for caffeine, phenol and the caffeine/phenol mixture should be measured at 254 nm and the retention times correspond to tr. for each compound. The chromatogram for NaNO3 should be recorded at 210 nm. The retention time measured is tm. (vi)

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(viii) calculate tr(the adjusted retention time), k (the capacity factor), (the relative retention), N (the number of theoretical plate), H (the height equivalent to a theoretical plate) and R the resolution using the information supplied in the table.

Quantity Partition coefficient CH4304 Laboratories

equation
K= C
s

Cm

Parameters Cs = conc of solute in stationary phase 25

Adjusted retention time Capacity factor

t 'r =t r t m

Cm= conc of solute in mobile phase tr= retention time of solute of interest tm = retention time of unretained solute Vs = volume of stationary phase Vm = volume of mobile phase ts = time solute spends in stationary phase tm = time solute spends in mobile phase subscripts 1 and 2 refer to two solutes w = width at base w1/2 = width at half height = std deviation of band x = distance travelled by centre of band L = length of column N = number of plates on column tr = difference in retention times Vr = difference in retention volumes wav = average width at baseline N = number of plates

k' =t r ' / t m= KV s / V m k' = ts tm

Relative retention

t' k' K = r2 = 2 = 2 t' r1 k'1 K 1 16t 2 N = 2r w 5 . 55t2 = 2 r w 1/ 2

2

Number of Plates

Plate height Resolution

L = x N t V r R= r = wav w av H= N R= 4

k '2 1 1k 'av

= relative retention k 2' = capacity factor for second peak k 'av = average capacity factor

DETERMINATION OF NITROGEN BY KJELDAHL METHOD

The method
The Kjeldahl digestion converts nitrogen compounds (proteins, amines, organic compounds) into ammonia compounds. Free ammonia is released by the addition of caustics, which is then expulsed by distillation and subsequently titrated. Kjeldahl tablets or copper sulphate/selenium salts accelerate the reaction.

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The tablets consist of sulfate and metal salts. The sulfate salt raises the boiling point of sulfuric acid, while the selenium, coppers, titanium, or mercury salts shorten the digestion time. Digestion catalyst CHNO + H2SO4 sample CO2 + SO2 +H2O +(NH4)2SO4

Weigh out accurately part of the organic sample (1.5 - 2.0g). Add 10g of potassium sulphate, 0.2g selenium or 0.5g anhydrous copper sulphate and pour 25ml of concentrated sulphuric acid into the flask in such a way as to wash down any solid adhering to the neck. Prepare three sample flasks and one blank flask. Allow to digest under vacuum for 2 to 4 hours. The addition of some glass beads to the flasks before boiling speeds up the digestion process. When cool carefully add 100ml of water to each flask and transfer to the distillation apparatus.

Neutralisation H2SO4 + 2 NaOH (NH4)2SO4 + 2 NaOH Na2SO4 + 2 H2O Na2SO4 + 2 NH3 + 2 H2O

Add excess of 50% sodium hydroxide solution. Distill off the ammonia into excess standard acid or a saturated solution of boric acid.

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Collection of NH3 {H[B(OH)4] + NH3

NH3 + H3BO3 NH4[B(OH)4]}

NH4

+ H2BO-3

The borate formed is determined by titration with standard 0.1N HCl; one mole of HCl being required for each mole of NH3 Titration H2BO3 + H

H3BO3 (NH4)2SO4 + 2 H[B(OH)4]}

{2 NH4[B(OH)4] + H2SO4

The solution at the equivalence point contains H3BO3 and NH4CL, so an indicator in the pH range 5-6 is necessary. Bromocresol green or bromophenol blue.

Calculate the % Nitrogen in the sample. What methods would you use to reduce or eliminate determinate error from this experiment.

Karl Fischer Titration of H2O

Water can affect the chemical and physical properties of substances and influence the quality, price and the shelf-life of a product.

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Using the Karl Fischer titration, the water content of gases, liquids and solids can be determined easily and with a high degree of accuracy. Determination of water content according to Karl Fischer is rapid, accurate and reliable; it is often the method of choice in quality and in-process control, production, research and development. It is a very widely used procedure for measuring water in food and pharmaceutical samples. The Karl Fischer Reagent is comprised of iodine sulphur dioxide, a base and an alcohol. What is a Karl Fischer chemical reaction? I2 + 2H20 + SO2 2Hl + H2SO4

This reaction occurs in the presence of a base and a solvent (a typical solvent could be methanol or diethylene glycol, and a base imidazole) A chemical reaction takes place between iodine (I) and water (W) with the reactants being in a 1:1 ratio between (I) and (W). 1x (I) + 1x(W) --> Reaction The end-point is usually detected coulometrically using an auto-titrator. The anode solution contains an alcohol, a base, SO2, I-, and possibly another organic. Pyridine was the primary ingredient in Karl Fischer solutions but has mostly been replaced. The alcohol is diethylene glycol and the base imidazole. The anode generates I2 by oxidation of I- in the presence of H2O, A stoichiometric reaction occurs between the alcohol, (ROH) base (B), SO2, and I2.

B.I2 + BSO2 + B + H2O

+

BH+I- + +B-SO3-

B-SO3- + ROH BH+ROSO3The net rx is oxidation of SO2 by I2 with the formation of alkyl sulphate. One mole of I2 is consumed for each mole of H2O. Once the iodine in the Karl Fischer reagent is determined, the unknown concentration of water in the sample can be determined. ie, as 1 molI2 reacts

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with 1 mol H2O, the amount of (I2) used during the titration must be equal to the unknown amount of water present in the sample. Coulometric Karl Fischer: - With a coulometric Karl Fischer titration, the amount of water present is determined by measuring the amount of current generated during the titration (coulombs) Coulombs are a measurement of current (amps) multiplied by the titration time in seconds. There is a relationship between the iodine (I2) used in the titration, the samples water content (W) and the current. ie, Current (A) x time (secs) = Coulombs (C) ---> I + W ---> KF reaction According to Faradays Law :- 2 x 96,485 Coulombs are needed to generate 1 mole of iodine and this iodine subsequently reacts 1 : 1 with the water in the KF reaction. How does a coulometric titration work? Coulometric KF titrations are preferably carried out within the pH range 47 Stage 1 Iodine generation Instead of dispensing KF reagent as in Volumetric KF titration, the Metrohm KF instrumentation actually generates the reagent inside the reaction cell. A current flows through the reagent generating iodine at the anode electrode. Stage 2 The instrument detects the end of the titration (end-point) Stage 3 T he instrument subsequently calculates the moisture content. Determine the moisture content of the sample provide quotingthe average and standard deviation.

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