SIC3006/SID3006 Practical Class (2021/2022, Semester 1)

QUIZ

Title: Analysis of Trace Metals in Milk by Flame Atomic Absorption

Spectrophotometry (FAAS) (https://youtu.be/Mwan7nJ30cY)

Name : Muhamad Nabil Najmuddin Bin Azman

No Matrik : 17188965/2

A. Standard Calibration

1) Please explain in details step A to E.

Step A : Pre-rinse a volumetric flask (of appropriate volume) 3 times using ultra
clean water. Keep stoppers on flasks except when adding water or reagents. Never
put stoppers ‘nose’ down on the table

Step B : Add ultra clean water (or matrix solution) into the volumetric flask
(approximate 2/3 full). This can help reduce analyte ‘sticking’ to glass surface.

Step C : Pipette appropriate volume of standard solution into volumetric flask being
careful not to contaminate pipette tip

Step D : Add ultra clean water (or matrix solution) until just below the meniscus line.
Use a dropping pipette to make up to volume: bottom of the meniscus of the solution
should be on meniscus line

Step E : Put cap on volumetric flask and then invert and rotate 10-12 times to
thoroughly mix solution. Ensure bubble in neck is completely displaced each time

2) What is the importance of step B, D and E?

Step B - Adding water before the analyte can reduce analyte ‘sticking’ to glass
surface on the bottom.

Step D – By using dropper, we can estimate the amount of water that needed so that
it does not exceed the meniscus level and also reduce the error.Basically it is more of
the same except to prevent the analyte from sticking to the top of the flask, and also
to make sure the volume is exactly as intended
Step E – We must invert and rotate the volumetric flask to make the solution become
homogenous which means that we want to make sure the solution mix is uniform and
the analyte is evenly distributed so that reaction can be happened.

3) What is the importance of standard calibration in analytical analysis?

Calibration standards are key to the analytical process. They are used to create the
calibration curve and the better the calibration curve, the better the data. Almost all
analytical methods require calibration with chemical standards.Mainly it helps to
obtain a valid relationship between the signal produced by the analyte and the
quantity of analyte in two or more samples of known analyte concentration
(standards), that can be described mathematically and used by the analyst to obtain
quantitative information on the analyte in samples of unknown analyte concentration.
Besides, systematic errors also can be eliminated by calibration.

And since the concentration of the standard calibration solution is known, we can
minimise measurement errors by making sure the equipment is accurate and precise.

B. Sample Preparation

1) Why do we need to prepare the samples in triplicates?

To make sure the data is accurate, as having near-similar results in threes allows us
to find the proper range of measurements, and thus find a good average for an
accurate result of the experiment. Later replications of the experiment can thus be
ensured to have the same results (unless two or all three of the samples have errors
in their analysis).

As we know collecting and analyzing triplicate samples allows for statistical

calculation of several important quantities including the standard deviation, RSD, and
95 percent (%) upper confidence level (UCL) of the mean. Which means, assaying
the samples in triplicate is another form of control. If we do not get the same result in
all three, could be there are problem with the experimental technique or there is error
in pipetting techniques. In a clinical laboratory, the experiment would have to be
repeated.

2) What is the purpose of sample filtering prior to HPLC analysis?

Among important reasons for using filtration during HPLC sample preparation are to
protect the pump from particulates in the solvent or released from rubber
components. Besides that is to prevent particulates clogging and damage the
delicate HPLC column.
C. FAAS

1) Explain why is FAAS a good method of quantifying Ca in milk?

AAS is a relatively inexpensive, simple, and safe method for determining the metal
content of solids and liquids that produces appreciably good results. For FAAS, it is
also suitable for the measurement and analysis of elements present in biological
fluids (in this case, Ca in milk).

2) How do you know if the milk samples are well digested / prepared with a
microwave digester? What should be done if they are not?

To know if the the milk sample is well digested is by look the techniques that is often
used for digestion which is ashing, which requires burning of the sample matrix until it
becomes ash. The ash is usually soluble in an acid solution. Other than that, the
other indicator is the colour and look of the sample should be clear and not muddy. If
they are not well digested, add an extra of nitric acid and hydrogen peroxide.

3) Why do I have to prepare a blank sample for Ca analysis using FAAS?

This blank allows for any metal concentrations present in the reagents (including
dilution water) to be accounted for and subtracted from the metal concentrations
found in the sample.Meaning to say that we are making sure there are no
contamination caused by errors during the experiment. It is a reference point to
compare with the actual reaction caused by the experimental procedure with other
samples. It does not contain matrix such as distilled water.

4) Why should we start the standard with the lowest concentration when
analyzing the Ca standard using flame AAS?

To determine the sample's detection limit. The lowest concentration will be the one
that can be distinguished clearly differentiated from zero for precision purposes.

5) In the analysis of Ca in milk samples, why is a recovery study essential?

Recovery studies are necessary for estimating the proportional systematic error of
the prior experiment as a whole. If there are any flaws, the analytical method used
was either incorrect owing to systematic error, or it was inaccurate due to random
instrument errors, or the experiment was contaminated, in which case the suspected
contaminant will be tested as well if known.