Calculation - Lab A. Standard Solution: m1v1 m2v2
Calculation - Lab A. Standard Solution: m1v1 m2v2
CALCULATION – lab
a. Standard solution
m1v1 = m2v2
m1 - concentration of stock solution
v1 - volume of stock solution
m2 - final concentration of new solution
v2 - final volume of new solution
m1v1 = m2v2
(1000) (V1) = (0.01) (0.1)
V1 = 1 µL
m1v1 = m2v2
(1000) (V1) = (200) (0.1)
V1 = 0.02 L
Standard solution
m1v1 = m2v2
(200) (V1) = (0.01) (0.1)
V1 = 5 µL
b. Dilution factor
𝑓𝑖𝑛𝑎𝑙 𝑣𝑜𝑙𝑢𝑚𝑒
Dilution factor =
𝑠𝑜𝑙𝑢𝑡𝑒 𝑣𝑜𝑙𝑢𝑚𝑒
𝑓𝑖𝑛𝑎𝑙 𝑣𝑜𝑙𝑢𝑚𝑒
Solute volume =
𝑑𝑖𝑙𝑢𝑡𝑖𝑜𝑛 𝑓𝑎𝑐𝑡𝑜𝑟
= 30 mL
c. Concentration in elements
i. River water
𝑓𝑖𝑛𝑎𝑙 𝑣𝑜𝑙𝑢𝑚𝑒
Dilution factor =
𝑠𝑜𝑙𝑢𝑡𝑒 𝑣𝑜𝑙𝑢𝑚𝑒
100𝑚𝐿
=
50𝑚𝐿
=2
ADVANTAGES DISADVANTAGES
1. Losses due to retention to the ashing
1. The ability to decompose large sample sizes.
container.
2. The need for little or no reagents.
2. Losses due to volatilization.
3. The technique is relatively safe.
3. Contamination from the ashing
4. The ability to prepare samples containing
container.
volatile combustion elements such as sulfur,
4. Contamination from the muffle furnace.
fluorine and chlorine (the Schöniger oxygen
5. Physical loss of 'low density' ashes when
flask combustion technique is very popular in
the muffle door is opened (air currents).
this case).
6. Difficulty in dissolving certain metal
5. The technique lends itself to mass production
oxides.
ADVANTAGES DISADVANTAGES
1. Almost no losses of analytes
2. Minimal contamination 1. Some materials are particularly resistant
3. Much safer than decomposition using HClO4 to acid digestion, e.g., certain rocks,
4. No difficulties associated with the use of H2SO4 mineral oxides, phosphates, and some
5. Low consumption of acids iron alloys
6. Fast decomposition (microwaves) – automation 2. Introduction of additional salts into the
7. Quicker method final solution
8. No difficult dissolution of ashed materials
10. WHEN DO WE USE HYDROFLUORIC ACID OR PERCHLORIC ACID FOR DIGESTION? - lab
Hydrofluoric acid is used in industry to digest to digest silicates in ore samples. In organic sample digestion,
nitric acid is rarely used alone. It is best used in the combination with sulphuric and/ or perchloric acid for
organic sample digestion. For samples that are not highly aromatic and/or contain a high -OH functionality,
nitric acid is preferably used followed by perchloric acid. The only element that may be lost from a
nitric/perchloric digestion is Hg
12. DISCUSS THE FUNCTION OF USING ACID HYDROCHLORIC ACID AND NITRIC ACID DURING THE
DRY ASHING PROCESS - lab
The function of acids in dry ashing is to dissolve the residue. Concentrated hydrochloric acid dissolved
many metals and forms oxidised metal chlorides and hydrogen gas. HCl is used in order to make some
organic compounds soluble in water. This compounds go through reactions to take in the chloride ion.
This makes the organic compound more polar by introducing a highly electronegative atom, which is
chlorine. Nitric acid can oxidise non-active metals
13. SOXHLET VS SHAKE FLASK EXTRACTION
14. HPLC VS GC
Non-volatile compounds can be easily analysed on HPLC whereas GC is useful for analysis of
volatile compounds
In GC, the mobile phase is a gas whereas in HPLC the mobile phase is liquid.
HPLC column are shorter and wide in comparison to GC column.
HPLC requires higher operating pressures than GC because liquids require higher pressures
than gases for transport through the system.
HPLC is useful for analysis of samples, which decompose at higher temperatures.
HPLC can be used for analysis of very high molecular weight compounds.
CRITERIA HPLC GC
Use liquid as carrier
Use gas as carrier
MOBILE PHASE Mixture of solvent of compatible
Single high purities gas
polarities
Separations carried out at
Separation carried out at ambient
elevated temperature
OPERATING temperature
Can be held at constant value
TEMPERATURE Packing shave extended temperature
or variables as decided to temp.
limit
program
Compound ranging in molecular
Compound separated have higher
weight up to few hundred
NATURE OF molecular weight ranging from a few
Such compound separate on
COMPOUNDS hundred to several millions for large
differences in their volatilities &
polymer & biomolecules
remain stable at high temp.
COLUMN Much longer & narrower
Much shorter & wider dimension
DIMENSION dimension
Higher permeability of solid
Offer greater resistance to flow of
supports than liquid
COLUMN liquids
The retention mode depends
PACKING The retention mode depends on
on difference in volatilities of
polarity differences/molecular size
compounds
Based on non-destructive detection
DETECTION such as UV, EI, photodiode array Based on destructive principle
PRINCIPLE detectors, conductivity, laser such as FID, NPD, FPD
detection
Solvent are costly
Maintenance cost of HPLC system Solvent less costly
COST OPERATION
higher due to pressure develop in Less maintenance cost
column & pump
Mobile phase
o As the name suggests High Performance Liquid Chromatography uses a liquid mobile phase and
gas chromatography uses a gas as the carrier. Liquids are generally mixtures of solvents of
compatible polarities whereas in gas chromatography the mobile phase is a single high purity
gas.
Operating temperature
o HPLC separations are mostly carried out at ambient temperatures whereas Gas
Chromatography separations are carried out at elevated temperatures which can be held at a
constant value (isothermal) or variable as decided by the temperature program. Newer packings
have extended temperature limits of High Performance Liquid Chromatography operation as
well.
Nature of compounds
o Gas chromatography separations are mainly carried out on compounds ranging in molecular
weights up to a few hundreds. Such compounds separate on differences in their volatilities and
remain stable at high temperatures. On the other hand compounds separated on HPLC have
higher molecular weights ranging from a few hundreds to several millions for large polymers and
biomolecules. Such compounds can be analysed at room temperature only because at elevated
temperatures they tend to degrade.
Column Dimensions
o Liquids used as carrier in HPLC generally have higher viscosity in comparison to gases used in Gas
Chromatography. This results in increased column back pressures in HPLC. It is for this reason
that High Performance Liquid Chromatography columns are much shorter and have wider
diameters in comparison to GC columns which can be much longer and narrower. Increased
column length improves resolution between closely spaced peaks. As the trend is towards faster
analysis columns used for HPLC are as short as 1 cm in length.
Column Packings
o Column packings offer greater resistance to flow of liquids in comparison to gases. Gases also
have a higher permeability of solid supports than liquids. The retention mode of HPLC columns
depends on polarity differences or molecular sizes whereas GC separations are based on
differences in volatilities of compounds
Detection principles
o HPLC detection is commonly based on non-destructive detection such as UV, RI, photodiode
array detectors, conductivity and laser detection. On the other hand, Gas
Chromatography detection is based largely on destructive principles such as a FID, NPD and FPD.
Mass spectrometry detectors common to both LC and GC are destructive in nature.
Cost of operation
o HPLC solvents are costly in comparison to gases used for GC analysis. In addition to cost of
solvents the maintenance cost of High Performance Liquid Chromatography systems is also
higher due to high pressures developed in pumps and columns. GC analysis in comparison is less
costly and there is lesser maintenance cost.
15. MICROWAVE DIGESTION SYSTEMS OFFER SEVERAL DISTINCT ADVANTAGES OVER OPEN
DIGESTIONS
Biggest benefit is time saving. Simultaneous heating of 8 – 12 samples is possible and reaction times
are typically less than an hour in comparison to 5 – 12 hours or even more for open digestions.
Time taken for a digestion is dependent on temperature. Open digestions are limited to boiling points
of acids used whereas closed digestion systems can be operated at higher temperatures of 250
– 300OC
due to ability to withstand higher pressure.
Acid consumption is lower in microwave digestion
Microwave digestion systems offer greater extraction efficiency whereas in open system digestion
extraction can be incomplete depending on discretion of the analyst.
No exposure of analyst to corrosive acid fumes as compared open digestions
Loss of volatile elements such as Hg or Pb can take place in open digestion whereas in closed system
digestions loss of volatile elements is prevented.
A greater risk of contamination from external sources exists in open digestion whereas in closed
systems the risk is non-existent
Closed microwave digestion systems have only one disadvantage, namely, explosion and cracking of digestion
tubes due to simultaneous build – up of pressure along with increase of temperature. However, microwave
systems have inbuilt sensors for both temperature and pressure. The microwave power is automatically
controlled or shut-off when pressure reaches the maximum limit