PHAR 32 LAB NOTES

TTH 7:30-10:30AM
JACKLYN SAN GABRIEL
VOLUMETRIC ANALYSIS
 IS OFTEN REFERRED AS TITRATION
 TITRATION = is a laboratory technique in which one
substance of known concentration & volume is used to react
with another substance of unknown concentration
VOLUMETRIC ANALYSIS
 Volume of a solution of known concentration consumed
during an analysis is taken as a measured of the amount of
active constituent in a sample being analyzed
TITRE
 Volume of titrant required to just completely react with the
analyte
ESSENTIALS IN TITRATION
1. TITRANT = volumetric or standard solution of known
concentration
2. ANALYTE = the active constituent to be analyzed; the
TITRAND
3. INDICATORS = chemicals capable of changing at or near
end point
 END POINT EQUIVALENT POINT
The point at which an equivalent or
The point at which the reaction is
stoichiometric amount of titrant is added
observed to be completed
to the analyte
The end point signal frequently
occurs at some point other than the
The point at which the reaction is
equivalent point.
complete
- tells the analyst to stop adding
TITRANT and record the volume.
Theoretically at the equivalence point we
The selected indicator should
can calculate the amount of titrant that is
change color very near to the
required to react EXACTLY with the
equivalent point.
amount of analyte present.
 Process – The Setup
 The burette is attached to a clamp stand
above a erlenmeyer flask
 The burette is filled with one of the
solutions (in this case a yellow standard
solution)
 A pipette is used to measure an aliquot of
the other solution (in this case a purple
solution of unknown concentration) into
the flask
 Prepare a number of flasks for repeat tests
 Last, an indicator is added to the
erlenmeyer flask
 Process – The Titration
 Read the initial level of liquid in the burette
 Turn the tap to start pouring out liquid of the
burette into the flask. Swirl the flask
continuously. When the indicator begins to
change colour slow the flow.
 When the colour changes permanently, stop the
flow and read the final volume. The volume
change needs to be calculated (and written
down). This volume is called a titre
 Repeat the titration with a new flask now that
you know the ‘rough’ volume required. Repeat
until you get precise results
PIPETTE
 For measuring accurate and precise volumes of solutions
 Used to transfer accurately, known volumes of liquids from 1
container to another
 TO DELIVER
 Should never be drained by blowing into them unless they are
especially graduated for use in this way
 The liquid is drawn up by using a commercially available
pipet filler = PIPETER
OSTWALD-FOLIN PIPETTE
 A special pipette used in measuring viscous fluids such as
whole blood
 It is calibrated to deliver (TD) by blowing out the last drop
MOHR PIPETTE
 A kind of graduated pipette
 Backward or drain-out pipettes have a 0mL mark just above
the end of the pipette
 A volume is pulled into the pipette, sometimes to the
maximum volume, then the needed volume is aliquoted out
SEROLOGICAL PIPETTE
 Blow-out pipette
 Have no 0mL mark as that corresponds to an empty pipette
 The volume needed is pulled into the pipette & then
dispensed
MOHR

BACKWARD

FORWARD

SEROLOGICAL
BURETTE
 For pouring measured volumes of solutions
 Used to dispense known amounts of a liquid reagent in
experiments for which such precision is necessary, such as
titration
 Graduated glass tubes of uniform bore throughout the whole
length, used in the measurement of variable quantities of
liquid
 They are closed at the bottom by a glass or Teflon Stopcock
to control the outflow of liquid
 PREPARATION
AND
STANDARDIZATION OF
1 N H2SO4
WEIGH 1.5 G OF PREVIOUSLY HEATED ANHYDROUS NA2CO3

DISSOLVE IN 100ML OF H20

ADD 2 DROPS OF METHYL RED = YELLOW

ADD H2SO4 SLOWLY FROM BURET WITH CONSTANT

STIRRING UNTIL SOLUTION BECOMES FAINTLY PINK

HEAT SOLUTION TO BOILING, CONTINUE TITRATION

PERSISTENT FAINTLY PINK COLOR = END POINT

STANDARDS
 A reagent solution of accurately known concentration is
called a STANDARD SOLUTION
PRIMARY STANDARD
 Is a highly purified compound that serves as a reference
material in all volumetric/titrimetric method
PRIMARY STANDARD
1. It should be 100% pure
2. Should be stable to drying temperature
3. Should be readily available
4. Should have high formula weight(to minimize weighing
error)
5. Should possess the properties required for a titration
(soluble and reacts readily)
ACIDIMETRY
 The direct or residual titrimetric analysis of bases using an
accurately measured volume of acid, is supplied in the
analytical control of considerable number of official
substances, both inorganic and organic
ACIDIMETRY
 DIRECT TITRATION = is conducted by introducing a
standard acid solution gradually from a buret into a solution
of the base being assayed until chemically equivalent amounts
of each have reacted as shown by some change in properties
of the mixture
ACIDIMETRY
 The END POINT, which must be close to the stoichiometric
point, is made evident by a change in the color of some
indicator or by potentiometric means
ACIDIMETRY
 Analysis of bases using an accurately measured volume of acid
ACIDIMETRY
 STANDARD ACID SOLUTION USED ARE USUALLY:
1. HCl = preferable to H2SO4 in the titration of compounds
that yield a precipitate with H2SO4 like Ba(OH)2
2. H2SO4 – preferable in hot titrations since there would be
danger of loss of HCl due to volatilization
ACIDIMETRY
 STANDARD AKALI SOLUTION COMMONLY USED:
1. NaOH May become contaminated w/ carbonates,that impair their usefulness
because after liberation of CO2 during acid-alkali titration
2. KOH
3. Ba(OH)2 – remain free from carbonates, since absorbed
CO2 is precipitated as insoluble barium carbonate, this
however decreased the conc. Of Ba(OH)2 in the solution
= alkali solutions should b prepared carbonate-free & should be
protected from CO2
= should be restandardized frequently
 What precautions should be taken when carrying
and pouring from a bottle of concentrated sulfuric
acid????
= always add sulfuric acid to the diluent and exercise great
caution.
= it is corrosive to all body tissues, so it must be handled
properly
= always add acid to water, not the other way around
 What emergency treatment is appropriate it
sulfuric acid contacts with the skin?
= irrigation with large quantitites of water for 10-15 mins to
cool tissue surrounding the acid burn and to prevent
secondary damage
= contaminated clothing must be removed immediately and the
underlying skin washed thoroughly
 The eyes?
= immediately flush with large amounts of water
 What are the pharmaceutical uses of sodium
bicarbonate?
= used in the manufacture of effervescent salts, baking
powders, fire extinguishers, carbonated drinks, and cleaning
mixtures
THERAPEUTIC USES:
1. Antacid
2. Combats systemic acidosis
3. Used in the treatment of methyl alcohol poisoning
4. Used in the management of sickle cell anemia
5. Laxative
 Write the equations of the reactions involved in the
standardization of sulfuric acid and assay of sodium
bicarbonate

STD:
Na2CO3 + H2SO4  Na2SO4 + H20 + CO2

ASSAY:
NaHCO3 + H2SO4  Na2SO4 + 2H20 + 2CO2
ASSAY OF SODIUM BICARBONATE
 Weigh 3g NaHCO3

Mix with 25ml distilled H2O

Add methyl orange TS = yellow

Titrate with 1 N H2SO4

End point = FAINT PINK COLOR

 How should standard solutions of sulfuric acid be
stored so that they will retain their strength?
= standard solutions of sulfuric acid preserved in tightly
stoppered, alkali-free bottles do not deteriorate
 Primary standard Na2CO3 was heated at 270 C for 1 hour
prior standardization to remove moisture.
What is effect to the results of analysis if sodium carbonate
contains varying amounts of moisture?
= sodium carbonate usually contains varying amounts of moisture and
sodium bicarbonate.
When a sample is heated to 270 C, the moisture is driven off and the
NaHCO3 is converted to Na2CO3

NaHCO3  Na2CO3 + H2O + CO2

 During standardization, why is the solution heated after the
first appearance of the pink color?
= the carbon dioxide generated as a result of the neutralization reaction
renders the solution acidic and introduces a source of error if not
removed.
Hence after the 1st end point is reached the solution is boiled to rid it of
C02
CO2 comes in contact with H2O, carbonic acid (H2CO3) is formed, affect
the pH & the pH or renders the solution acidic even if not yet the end
point
 COULD 1 N ACETIC ACID BE USED TO TITRATE THE
BICARBONATE IN PLACE OF SULFURIC ACID?

= 1N acetic acid cannot replace sulfuric acid because acetic acid is a

weak acid and will not dissociate completely. No sharp END POINT.
Reaction will not proceed to completion, as described by the equation:
CH2COOH + NaHCO3 CH3COONa
 WHAT ARE THE GUIDELNES FOR CONTENT
UNIFORMITY OF TABLETS? WHAT ARE THE ACCEPTABLE
LIMITS SET BY THE USP FOR NaHCO3 TABLETS?
= for content uniformity by the USP METHOD, 10 units are
individually assayed for their content according to the method
described in the individual monograph.
The requirements for content uniformity are met if the amount of
active ingredients in each dosage unit lies within the range of 85% -
115% of the label claim & relative std. Deviation is less than 6 %
 ACIDIMETRIC ANALYSIS
 RESIDUAL TITRATION = is used whenever the end point of a
direct titration deviates appreciably from the stoichiometric points
for some reason
ie. When the basic sample is insoluble in water and the rate of its
reaction with the standard acid is relatively slow
In such a procedure, the basic sample is treated with an amount of
standard acid known to be in excess of a chemically equivalent
amount, and the excess acid is subsequently titrated with a
standard base
ACIDIMETRIC ANALYSIS
 The amount of the standard acid which is chemically
equivalent to the basic sample can be calculated simply by
subtracting, from the total amount of standard acid
employed, the amount of acid neutralized by the standard
base in the back titration
 PREPARATION
AND
STANDARDIZATION OF
1N NaOH
 WEIGHT 45G OF NaOH

DISSOLVE IN 950mL COOLED BOILED H2O

STAND THE SOLUTION OVERNIGHT

WEIGH 5 G OF POTASSIUM BIPHTHALATE

DISSOLVE IN 75 ML OF CO2 – FREE WATER

ADD 2 DROPS OF PHENOLPHTHALEIN TS = observe color

TITRATE WITH NaOH SOLUTION TO THE PRODUCTION OF

PERMANENT PINK COLOR = end point
 WRITE THE BALANCED EQUATIONS FOR THE
REACTIONS INVOLVED IN THE
STANDARDIZATION OF NaOH AND ASSAY OF ZnO.

KHC8H4O4 + NaOH  KNaC8H4O4 + H2O

ZnO + H2SO4  ZnSO4 + H2O

H2SO4 + NaOH  NaSO4 + H2O

 WHAT PRECAUTIONS SHOULD BE OBSERVED IN
THE PREPARATION OF NaOH SOLUTION?
= it should be handled with care, as it is corrosive to tissues
when it comes to contact with skin
When adding H2O to NaOH, the process should be in a slow
manner to avoid tremendous release of heat since NaOH
from exothermic solutions with H2O
 WHAT IS THE USE OF ZnO IN THERAPY?

= ZnO has a mild astringent, protective and antiseptic action

Used to treat dry skin and skin disorders and infections (acne
vulgaris, prickly heat, seborrhea, psoriasis)
Also included in vulvovaginal deodorant prepartions, dental
cements & fillings
 WHY DO WE DISSOLVE OUR ANALYTE PARTICULARY WITH
C02-FREE WATER?
=NaOH solutions should be prepared using carbonate/CO2- free water
because the purity of NaOH will be significantly affected by the presence
of carbonates when these carbonates are in the solution
They react with the analyte during standardization, hence inaccurate end
point is reached with leads to inaccurate determination of concentration,
and incorrect analysis
CO2/carbonate decreases the concentration of NaOH
ASSAY OF ZINC OXIDE
WEIGHT 1.5G OF UNKNOWN/ZnO + 2.5G OF NH4Cl

DISSOLVE IN 50mL OF 1N H2SO4 = PINK COLOR

GENTLE HEAT
ADD METHYL ORANGE TS = PINK

TITRATE WITH 1N NaOH

END POINT = YELLOW

CALCULATIONS
GEW
 Gram-equivalent weight
Defined as that weight in grams w/c is chemically equivalent to
1 gm-atom of Hydrogen

Weight of sample
MW/# rep. ions
GmEW
 Defined as GEW/1000
 Used more frequently in titrimetric calculations

Weight of sample 1000  meq = MW/2/1000

MW/# rep. Ions
Normality
 Should be expressed to the 4th decimal place
 Is a concentration expression & is defined as the # of
equivalents of solute per L of solution
 equiv/L or meq/mL

N = gram- equivalent solute

Volume of Solution ( Li)

gram-equiv: weight solute (g)

equivalent weight
 Equivalent weight:
eq. wt ( acid) = molecular weight of acid (gram/mole)
no. of H+ replaced

eq.wt. ( base) = molecular weight

no. of OH- reacted

eq. wt (salt) = molecular weight

total no. of + /-
Calculation of the % purity of a sample
determined by direct titration

Volume(N) (equivalent weight) x100= %

Sample weight or volume
Molarity
 # of moles of solute/ L of solution
 An expression of the concentration of a solution in terms of
moles/L solution

M = mole solute
Volume Sol’n ( Li)
TITER
 Defines as the weight of a substance/analyte chemically
equivalent to 1mL of a std. Solution
 Usually expressed in mg

Ex. 1mL of HCl will exactly neutralize 4mg of NaOH, the titer
is 4mg/mL
T=mg/mL
T = N x eq wt.
EXERCISE
1. What is the normality of a NaOH solution made by dissolving 20
grams of NaOH in qs water to make two liters of NaOH
solution?

2. Calculate the normality of a magnesium sulfate

solution made by adding 4.50 grams of the solute in
enough water to make 1500 ml magnesium sulfate
solution.

3. How many grams of NaOH would be needed to

prepare 1 gallon of a 1.2000 N NaOH solution?
4. How many gram equivalents of the sulfuric acid is present in
five liters of a 5N sulfuric acid solution?

5. How many liter/s of HCl solution can be prepared from a 5N HCl

solution which should contain 2.675 gram equiv HCl

6. How many grams of H3PO4 are in 175 mL of a 3.5 N solution of

H3PO4?

7. How many grams of NaOH (MW = 40.0) are there in 500.0 mL of

a 0.175 N NaOH solution?