A labguide has been prduced for each experiment. It gives info on
preparations, chemicals, apparatus and sample requirements. It also
includes a helpful analytical notes feature.
To get the labguide for this experiment click here Enjoy!
Revised labscript :
1. Introduction:
Iron in pharmaceutical preparations may be provided in the form of ferrous sulphate, gluconate or ferrate salts.The active
ingredients are normally held in a matrix which must be destroyed, before the iron can be accurately determined. In this
experiment, the matrix is destroyed by oxidation with concentrated nitric acid. The iron, after conversion to the ferrous state, is
complexed with 1:10-phenanthroline and determined by visible spectrophotometry (1)
2. Experimental procedure:
(A) Sample Preparation:
A ground tablet sample will be provided for analysis.
(i) Weigh about 0.1g of the ground sample into each of three clean, dry boiling tubes.
(ii) Add to each of these tubes, plus one blank tube, 5 mL analytical grade nitric acid. Allow to stand in a fume hood overnight
(10-12h) to predigest.
(iii) Heat the tubes on a sand bath or heating block at 135-140°C,for 2-3h, to complete the extraction procedure.
(iv) Cool to room temperature, dilute the contents with 10mL distilled water and mix well. Filter through Whatman #1 filter paper
into 250mL volumetric flasks and make up to the mark with distilled water washings of the boiling tubes and filter
papers.
(v) Pipette out 2 mL of each sample solution into a 100mL volumetric flask. Add 5mL of 10% (w/v) aqueous hydroxylamine
hydrochloride solution and 5mL of 1M sodium acetate-acetic acid buffer (pH 5.0).
Add 4mL of 0.25% (w/v) aqueous 1,10-phenanthroline and make up to the mark with distilled water. Allow to stand for one hour
before making measurements on these solutions.
(vi) Similarly prepare a series of Fe standards (25mL each) containing 0, 1.0, 2.0, 4.0, 6.0, and 8.0ug Fe/mL respectively,
using the standard Fe solution provided.
(c) Visible Spectrophotometry:
(i) Switch on and allow spectrometer to warm up and stabilize.
(ii) Rinse and fill a clean cuvette with some of the standard blank solution and wipe the external surfaces clean with a soft
tissue.
Fill another cuvette with the 1.0ug/mL Fe standard solution.
(iii) Place the cuvettes in the reference and sample compartments respectively of the double-beam uv/vis spectrophotometer. If
a diode-array spectrometer is used, the blank sample is scanned first and the spectrum stored, for correction of the
absorbances of the sample solutions.
(iv) Scan the solutions over a wide wavelength range (see Demonstrator) and locate the the wavelength at which maximal
absorbance occurs, and the absorbance value at this wavelength. All subsequent measurements can be made on a single
beam instrument at this wavelength.
(v) Measure the absorbances of all the other standard and sample solutions at the chosen wavelength, corrected for their
respective blank values.
(c) Calculations:
Plot a calibration curve of Absorbance vs Fe concentration of the iron-phenanthroline complex and determine the Fe
concentrations of the sample solutions.
Alternatively, calculate the linear regression equation and correlation coefficient for the calibration curve and hence determine
the Fe concentrations of sample solutions.
Hence calculate the mean iron concentration of the sample provided ± standard deviation.
The Fe concentration of the sample will be provided (see Demonstrator). Do your results of analyses differ significantly (p<.05)
from the reference value provided?
(d) Exercises:
(i) Give the rationale for:
- preparation of blanks for samples and standards;
- scanning both sample and standard to establish λ max
(ii) How may the efficiency of this method be improved, in terms of:
- speed of analysis;
- economy of use of reagents?
Reference:
(1) Vogel's Textbook of Quantitative Inorganic Analysis 4th. edition (1988). Longman's Publ. 742 - 743.
anal-chem resources
Chem. Dept. UWI. St. Augustine Campus
Original labscript :
Determination of iron by UV-VIS spectrophotometry
Basic Theory
Iron in pharmaceutical preparations is normally in the form of ferrous
sulphate, gluconate or ferrate respectively.The active ingredients are
normally held in a matrix which must be destroyed before before the
iron can be effectively determined. In this experiment, the matrix is
destroyed by oxidation with concentrated nitric acid. The iron, after
reconversion to the ferrous state, is complexed with
1:10 - phenanthroline and determined by visible spectrophotometry. (1)
Sample Preparation
Place one tablet into each of five clean boiling tubes. Add to each
of these tubes, plus one blank,5 mls analytical grade conc. nitric
acid. Allow to stand in a fume hood overnight to predigest. After this
period, heat the tubes on a sand bath or heating block at 120-130°C
to complete the digestion procedure (2 hours). Cool and then dilute the
contents with 10 mls distilled water. Filter through a Watman #1 filter
paper into 250 mls volumetric flasks and make up to the mark with
distilled water washings of the boiling tube and filter paper.
Pipette out 2 ml each of your diluted solutions into 100 mls volumetric
flasks. To each flask add 5ml of 10% (w/v) aqueous hydroxylamine hydrochloride solution and 5 ml of 1M sodium acetate-acetic acid buffer
(pH 5.0), then add 4 ml of 0.25% (w/v) aquoeus o-phenanthroline and make up
to the mark with distilled water. Allow to stand for one hour before
making measurements on these solutions.
Similarly prepare a series of standards (25 mls) containing 0, 1.0,
2.0, 4.0, 6.0, and 8.0 ppm from standard stock solution provided.
Visible Spectrophotometry
Rinse and fill a clean cuvette with some of the standard blank solution
and wipe the external surfaces clean with tissue. Fill another cuvette
with standard. Place the cuvettes in the reference and sample
compartments respectively of the double-beam uv/vis
spectrophotometer.Scan between 480 and 520nm and note the wavelength
range over which maximum absorbance occurs. Repeat the procedure using
a sample / sampleblank pair to confirm the λ max
range.
All subsequent measurements should be made on a single beam instrument.
Set the mid point of the λ max on the single beam
spectrophotometer and measure the absorption values of each standard
and sample against its repective blank.
Plot a calibration curve and determine the linear regression equation
for the iron-phenanthroline complex and determine the concentration of
iron in each solution. Hence calculate the total iron content of each
tablet analysed, and then the mean iron content per tablet of the batch
± standard deiation by each method.
Assume that the tablets each contain x mg of iron.Do your results
differ significantly from the above amount at (a) the 95% (b) the 99%
confidence level?
Give the rationale for
(a) Preparation of blanks for samples and standards.
(b) Scanning both sample and standard to establish λ
max
List sources of error that effect the accuracy
and precision of analysis in this experiment.
Reference:
(1) Vogel's Textbook of Quantitative Inorganic Analysis
4th.
edition (1988). Longman's Publ. 742 - 743.
anal-chem resources
Chem. Dept. UWI. St. Augustine Campus
|
