UNIT I. INTRODUCTION TO PHARMACEUTICAL 4.

Special methods – those which require a distinct type of

ANALYSIS technique, such as alkaloidal assaying.
OBJECTIVES Official assay methods serve as an exact measure of the
purity of a substance only when the results are considered in
• Describing the different terms used in conjunction with the qualitative tests.
pharmaceutical analysis
• Identifying the parts of a product monograph A pharmacopoeia is a legally binding collection of standards
• Identifying the different analytical methods of and quality specifications for medicines used in a country or
analysis region. ... The role of a modern pharmacopoeia is to furnish
• Discussing the different parameters used in quality specifications for active pharmaceutical ingredients
analytical method validation (APIs), FPPs and general requirements.

QUANTITATIVE PHARMACEUTICAL CHEMISTRY SUCCESS AS AN ANALYST

It is the application of procedures of quantitative analytical • Analytical chem is not simple routine procedure.
chemistry: • Manipulative skill is required in analytical
procedures.
1. To the analysis and determination of the purity and • Acquires fundamental knowledge of the theory and
quality of drugs and chemicals used in the ability in the application of analytical methods
pharmacy, esp in the USP-NF • Must know what reactions are taking place during
2. To the analysis of the chemical constituents found
an analysis
in the human body whose altered concentrations
• Able to understand and apply the theory upon
during disease states serve as diagnostic aids in
which the method is dependent
the practice of medicine
• Must acquire skill of technique, patience, neatness,
3. To the analysis of medicinal agents and their
and accuracy
metabolites found in biological system
• Bear in mind that analysis is used as a means of
QUALITY AND QUALITY CONTROL controlling the purity and strength of substances
used as medicaments.
• As applied to the drug product, it is the sum of all
factors which contribute directly or indirectly to the ACCURACY AND HONESTY
safety, effectiveness, and reliability of the product.
• All quantitative work be performed without a slight
• QC: guarantees within reasonable limits that a drug
loss of material or gain of extraneous matter, take
product
every precaution
1) Is free of impurities
• All precipitates and solutions should be covered to
2) Is physically and chemically stable
protect from dust
3) Contains the amount of active ingredient(s)
as stated on the label • As analyst, the only person familiar with the entire
4) Provides optimal release of active history of analysis and knows whether it is worthy of
ingredients when the product is full confidence
administered. Of the four properties listed, NOTEBOOKS
item 3 is usually emphasized in quantitative
pharmaceutical chem • The following information relative to each determination
should be included;
METHODS OF ANALYSIS
o Date – very impt
1. Volumetric analysis – is the determination of the volume of o Object or title- should identify the sample and
a solution of known concentration required to react with a analytical value measured
given amount of the substance to be analyzed. o Experimental data-complete record of all weighings
2. Physicochemical methods – are based on some specific & measurements
physical or chemical properties of a substance being o Reactions- should be given ,using structural
analyzed, it employs instrumental techniques (chromat, formulas where possible
spectrophotometry). o Calculations - calculator is highly recommended
o Results – should always be reported as percentage
3. Gravimetric analysis – it is separation by extraction,
o Remarks – include any unexpected development
precipitation, or other means, of the constituent to be
during analysis, explain errors, and comments
determined, and weighing the resulting product.
This information should be arranged in a neat, systematic c. Apparatus errors due to poor calibration
manner so that anyone familiar with quantitative analysis (ex.incalibrated pipets)
may understand it readily.
ACCURACY AND PRECISION
ECONOMY OF TIME
• Accuracy – used to denote the agreement of an
• There is no such thing as “speed” in quantitative work. experimental result or the agreement of the mean value of a
Economy of time is achieved by planning the work to carry series of experimental results with the true value , and is
two or more operations at one time, otherwise, repeat a usually expressed in terms of error.
determination.
o The difference between the mean and the true
• To save time and increase accuracy: value is known as absolute error.
o The relative error is found by dividing the absolute
1. Keep the desk scrupulously clean at all times
error by the true value and expressed as %.
2. Clean all apparatus immediately after using it
3. Label all solutions, filtrates, and precipitates • Precision – is a measure of reproducibility of data within a
systematically series of results. Results within a series which agree closely
4. Keep 2 or more operations going at one time with one another are said to be precise.
(ex.weighing while incinerating)
5. Utilize all time between operations in making o Precision is usually reported as the average
calculations and writing up experiments. deviation, standard deviation, or range.

• Reagents should conform to the requirements of USP-NF. • Average deviation (d) is calculated by finding the difference
A “reagent-grade” will usually be found to meet all official between individual results and the mean, regardless of sign,
requirement. adding these differences, and dividing by the number of
determinations.
• Purity and strength requirements are usually expressed in
terms of percent. The official requirements of purity and o Relative average deviation is found by dividing the
strength pertain only when the official methods of assay are average deviation by the mean, usually expressed
employed. in % or as parts per thousand.

SAMPLING • Standard deviation (s) is calculated using the equation and

is the preferred measure of precision.
• The first important considerations in an analysis is
the securing of a representative sample. • Relative standard deviation sometimes called coefficient of
• A sample from each package should be examined variation, is found by dividing the standard deviation by the
separately, taken from the different parts of the mean and multiplying by 100.
container and mixed, then portion of this mixture • Range is the difference between the largest and smallest
should be taken for analysis. results in a series of measurements.
• Bulk quantities of chemicals and drugs may be
sampled by the methods given in the USP for EXAMPLES
sampling vegetable drugs.
Compute
CALCULATION OF RESULTS AND ERRORS
1. Mean : x̃= 0.2140 + 0.2142 + 0.2152 + 0.2140
Source and nature of errors 2. Range R
3. Ave deviation (d)
2 types: 4. Relative average deviation
1. Indeterminate errors – manifest by slight variations in a 5. Standard deviation (s)
series of observations made by the same observer under 6. Relative standard deviation
identical conditions SIGNIFICANT FIGURES
2. Determinate errors – it recur in a constant manner in each • Significant figures in a number are defined as all
of a series of determinations. certain digits of a measurement plus one doubtful
causes: digit.
• Zeros within a number such as 20.04 are always
a. personal errors by analyst (ex. Inability to judge significant
color changes) • Zeros used as shown in the number 0.043 are
b. Errors of method due to faulty procedure never significant, but show only the order of
(ex.contamination) magnitude of the other digits
 • When terminal zeros are significant, they are written
in the usual manner with the other digits of the
number.
• When terminal zeros are used to locate the decimal
point of a measurement, they are not significant and
it is recommended that the number be expressed
as a power of 10.
• •In general, for analytical work, the operations of
addition and subtraction should give no more
decimal places than the least number of decimal
places entering into the calculation.
• Ex. 12.4 + 121.502 + 3.6653 = 137.5673, the result
should be written 137.6, however, it is better to
retain one more figure, i.e. 137.57
SIGNIFICANT FIGURES IN PH CALCULATIONS • Proposing an optimized validation scheme The most critical
• For pH calculations, retain the same number of step in validating a method is to establish a protocol
significant figures to the right of the decimal as the containing well-defined procedures and criteria. Testing
total number of non-exponential significant figures validation parameters consumes time and resources;
of the hydronium-ion concentration. however, a well-planned and organized protocol results in a
• The pH of a 3.4 x 10 -12 M concentration of
hydronium-ion solution is expressed as pH = 11.47.
It is permissible to express all pH calculations to
two places to the right of the decimal.
DRUG MONOGRAPH

• It is a summary of the scientific findings of a drug

that provide information on appearance,
ingredients, effects, side effects, indications,
dosage, including details relating to medicinal areas
of use of a drug.
• The quality requirements for a drug and the testing
methods prescribed for the quality are defined in
the form of drug monographs in the USP-NF, BP,
etc.
rapid and concise method validation procedure.
METHODS OF VALIDATION

• To validate means to officially state that something

The following parameters should be considered in order to
that has been proven to be true is useful and of an
obtain the proposed scheme:
acceptable standard.
• The validation of analytical procedures intends to 1. Linearity
establish the performance characteristics of
analytical applications through experimental tests,  Linearity is the ability of obtaining results that are
resulting in a suitable analytical method for its directly proportional to the analyte concentration in
purpose the sample, across the specified range, i.e., the
interval between the upper and lower levels of
What to be considered in the validation study analyte.
 This test should be run first, since its data might be
• Analytical procedures might be validated for
used for assessing repeatability, intermediate
investigating the identity of the analyte in a sample
precision (IP), accuracy, LOD, LOQ, stability and
(identification tests);
system suitability.
• obtaining the analyte content (quantitative tests);
and 2. Precision
• analyzing impurities (quantitative or limit tests).
 Precision is the closeness of agreement between
individual results obtained from a repeatedly
 applied procedure in a homogeneous sample,  Concerning liquid chromatography, variations such
comprising repeatability and IP. as mobile phase composition, mobile phase pH,
 It is expressed as the standard deviation or column temperature, column lots, column suppliers
coefficient of variation. and flow rate might be set.
 It can be evaluated by the analysis of the standard
2.1. Repeatability
solution of analyte at 100% of expected
 Repeatability is defined as precision over a short concentration in triplicate for each isolated condition
interval of time under the same operating and expressed as the standard deviation and
conditions, i.e., same day, same analyst and same relative standard deviation.
equipment.
6. Limits of detection and quantitation
 It is obtained by analyzing standard solutions of
analyte at 3 different concentrations covering the  Both LOD and LOQ parameters can be directly
specified range in triplicate. obtained from the linearity test. The lowest amount
 As shown in the scheme, it can be directly obtained of analyte which can be detected under the stated
from the linearity test experimental conditions is the LOD.
 LOQ is the lowest amount of analyte which can be
2.2. Intermediate precision (IP)
quantitatively determined with precision and
 IP expresses within-laboratory variation, accuracy under the stated experimental conditions.
considering different days, different analysts or 7. Stability
different equipments.
 In practice, it is repeatability performed for 3 times.  Since chemical decomposition may occur during
Therefore, the first determination of IP can be storage, drug stability should be evaluated by the
obtained from the repeatability test; the second and analysis of sample and standard solutions during a
third determinations are obtained by repeating the preselected period of time, allowing the estimation
test for 2 more consecutive days. of the maximum interval between sample
preparation and analysis.
3. Accuracy
Vidwatch:
 Accuracy is the closeness of agreement between
test results across the specified range and an https://www.lucideon.com/healthcare/pharmaceuticals/pharm
accepted reference value. aceuticalquality-control-testing
 It is obtained by the standard addition method, in
which the sample is spiked with known quantities of
the analyte.
 In practice, it can be evaluated by spiking a sample
with the standard solutions of analyte at 3 different
concentrations used in the linearity test in triplicate.
 It is expressed as the percentage of analyte
recovery, standard deviation and relative standard
deviation.
4. Specificity

 Specificity is the ability to assess unequivocally the

analyte in the presence of substances that are
expected to be present, such as excipients,
impurities and components of the mobile phase.
 It might be assessed by verifying the absence of
interfering peaks at the analyte retention time when
spiking a sample with appropriate levels of
impurities or excipients.
5. Robustness

 Robustness is the capacity of the analytical method

to remain unaffected by small variations in its
parameters, indicating reliability.
 A. chemical  relies on chemical rxn
B. Physical  physical properties
QC 2: ANALYSIS
C. Biological use whole organisms or parts of
Objectives organisms as analytical indicators

 To demonstrate the different ways of expressing the ■ Reagents used in analysis:

strength concentration of a solution 1. Technical or commercial grade
2 divisions of Analytical Chemistry : 2. USP grade
3. Chemically pure or CP grade
 Qualitative 4. Analytical Reagent or AR grade/Laboratory grade
 Quantitative
■ Types of Solution Based on Concentration:
Classification of Analysis:
1. Concentrated Solution
1. Based on sample size 2. Dilute solution
3. Saturated solution
A. Macro
4. Unsaturated solution
B. Semi-micro
5. Supersaturated solution
C. Micro
D. Ultra Micro WAYS OF EXPRESSING STRENGTH OF SOLUTION
1. Weight/unit volume
2. specific gravity
Example:
A. What is the weight in grams of 10 ml H2SO4 whose
sp. gr. is 1.84?
B. What is the volume in ml of 12 grams HCl whose sp
gr is 1.15?
C. If 54.96 mL of an oil weighs 52.78 g, what is the
specific gravity of the oil?
D. A 50 mL pycnometer is found 120 g when empty,
171 g when filled with water, and 160 g when filled
with an unknown liquid. Calculate the sp. Gr of the
unknown liquid?
E. Get ¼ paper
ANSWER THE FF:
2. Based on extent of determination (ex.ash)
1. what is the weight, in kilograms, of 5 liters nitric acid
A. proximate -comprises the mass percentages of with specific gravity of 1.84?
moisture, ash, volatile matter, and fixed carbon, 2. what is the volume, in pints, of 22,700 g of glycerin
which are obtained from a series of three having a specific gravity of 1.25?
standardized tests. 3. A sp. Gr bottle weighs 23.66g. When filled with
B. ultimate- more comprehensive, is dependent on water, it weighs 72.95 g; when filled with another
quantitative analysis of various elements present in liquid, it weighs 73.56 g. what is the sp. gr of liquid?
the coal sample, such as carbon, hydrogen, sulfur,
oxygen, and nitrogen WAYS OF EXPRESSING STRENGTH OF SOLUTION

3. Based on nature of methods 3. Ratio

A. Classical - analytical techniques that use no 4. Percent strength

mechanical or electronic instruments other than a
Example:
balance.
B. Instrumentation – use of spectro,thermal 1. What is the % strength of a ½ liter NaCl solution
analysis,etc containing 8 grams of solute?
C. Miscellaneous/Specific method 2. How many ml of a 2% Iodine solution can be
prepared out of 22 grams iodine crystal?
4. Based on materials used
 3. How many grams of silver nitrate will you need to  Sp.gr = density of subs/density of water
prepare 1 liter of a 5% silver nitrate solution? 0.8 = x/1g.ml, x = 0.8g/ml
0.8g/ml = 225g/x, x = 281.25ml
Concentration Calculations
Qtty of solute = 281.25ml
1. Percentage weight-in-volume (w/v) CONC = qtty of solute /qtty of prepn
x/100ml= 281.25ml/1500ml
■ Expresses the number of grams of a constituent in 100ml X = 18.75% (v/v)
of solution or liquid preparation. Expressed as ____%w/v.
■ Ex. % w/w
2. Percentage volume –in-volume (v/v)
■ How many grams of hydrocortisone should be used in
■ Expresses the number of milliliters of a constituent in preparing 120 suppositories, each weighing 2 g and
100ml of solution or liquid preparation. Expressed as ____ containing 1% of Hydrocortisone?
%v/v.
Data:
3. Percentage weight-in-weight (w/w)
conc = 1% (w/w) = 1g/100g
■ Expresses the number of grams of a constituent in 100 g Qtty of prepn = 2g x 120 = 240g
of solution or preparation. Expressed as ___%w/w
Percentage v/v, w/w, and w/v CONC = qtty of solute / qtty of prepn
1g/100g = x/240g
 Concentration may be defined as the ratio of the x = 2.4g
quantity of a solute in quantity of preparation.
 Conc = quantity of solute/quantity of preparation Calculating %m/m or %w/w
A. 25 g NaCl in 65g H2O; %mass=?
■ Ex. %w/v
B. 75 g of a 12% (m/m) NaCl soln; g NaCl=?
■ A formula for antifungal shampoo contains 2% w/v C. 100g NaCl in a 25% (m/m) NaCl soln; g soln=?
ketoconazole. How many grams of ketoconazole would be
WAYS OF EXPRESSING STRENGTH OF SOLUTION
needed to prepare 25 liters?
ANSWER THE FF:
Data:
1. how many grams of aminobenzoic acid should be
 Conc=2%(w/v)= 2g/100ml
used in preparing 235 mL of 5% solution in 70%
 Quantity of preparation=25L x 1000ml/1L =
alcohol?
25,000ml
2. what is the percentage strength v/v of a solution of
 CONC = qtty solute/qtty of prepn
800 g of a liquid with a sp gr of 0.800 in enough
 2g/100ml = x/25,oooml water to make 4000 mL?
 X = 500g 3. if 5 g of boric is added to 100 ml of water, what is
■ How many ml of a 0.9% (w/v) solution of sodium chloride the percentage strength (w/w) of the solution?
can be prepared from 50 tablets, each containing 2.25g of WAYS OF EXPRESSING STRENGTH OF SOLUTION
sodium chloride?
5. Molar solution
Data:
Molarity (M) is the concentration of a solution expressed as
 Conc= 0.9% (w/v) = 0.9g/100ml the number of moles of solute per liter of solution: Molarity
 Qtty of solute = 2.25g x 50 = 112.5g (M) =mol solute/L of solution
 CONC = qtty of solute/qtty of preparation
 0.9g/100ml = 112.5g/x Examples:
 X = 12,500ml A. A solution is made by dissolving 2.355g of H2SO4, in
water. The total volume of the solution is 50.0 mL. What is
■ Ex. % v/v
the molar concentration of sulfuric acid?
■ What is the percentage strength (v/v) if 225 g of a liquid
■ To find [H2SO4], we need to find out how many moles of
having a specific gravity of 0.8 is added to enough water to
sulfuric acid are in solution. We can convert the mass of the
make 1.5L of the solution?
solute to moles using the molecular weight of sulfuric acid,
Data: 98.08 g/ mol:

 Qtty of prepn= 1.5L = 1,500ml Mol H2SO4=2.355g H2SO4 x 1mol/98.08g = 0.02401 mol
■ Then plug in the moles of sulfuric acid and total volume of Looking For
solution in the molarity equation to calculate the molar
Units- Equivalent Weight
concentration of sulfuric acid:
The equivalent weight of a compound is defined as
[H2SO4] = mol solute/L of soln
■ EW=Atomic or molecular weight/n
= 0.02401mol/ 0.050L
g/mol eq/ mol  g/eq
= 0.48 M
n - is the number of electrons or protons transferred in rxns;
WAYS OF EXPRESSING STRENGTH OF SOLUTION
1. If 28 grams of KOH are dissolved in q.s. water to - The number of hydrogen ions that a molecule transfers
make a liter solution, what is the molarity of the - the valence state of an element or compound
above solution? ■ What is the equivalent weight of the following?
2. How many grams of silver nitrate will be needed to
prepare 400 ml 0f 0,25 M solution? ■ EW=MW/n
3. Prepare 100 mL of 1.0 M HCl from concentrated
HCL (12.1 M)
4. How many mL of 12 M HCl are required to prepare
500 mL of 6 M HCl?
ANSWER THE FF:
1. Prepare 800 mL of 2M sodium chloride.
2. How many grams of NAOH will be needed to GRAM EQUIVALENT WEIGHT OF ACIDS AND BASES
prepare 2,000 ml 0f 0,15 M solution?
3. If 105 grams of NAHCO3 are dissolved in q.s. water  The GEW of an acid is that weight of it which
to make a 5,000 mL solution, what is the molarity of contains 1.008 gm of replaceable hydrogen.
the above solution?  The GEW of a base is that weight of it which
contains one replaceable hydroxyl group.
6. Normal solution : Units-Normality
ex. HCl and NaOH have only replaceable H and OH hence
Normality- the no. of equivalents weights of a solute their equivalent wts are equal to their mol.wt.
contained in 1L of a soln.
Mol wt of HCl=H (1x1) + Cl (35.5 x1)
A solution which contains 1gm equivalent of the solute per
liter of the soln is called normal, represented as 1N. = 1+35.5  36.5 gm/equiv

N=Mn Number of replaceable ions (H+) =1

GEW = mol wt/no.of RI
where: M=mol/L, n=eq/mol  N=eq/L = 36.5/1 = 36.5 gm
Where:
n - is the number of electrons or protons transferred in rxns ■ Mol.wt of NaOH= Na(23x1) + O(16x1) + H(1x1)
- The number of hydrogen ions that a molecule transfers -
= 23+16+1 40gm/equiv
the valence state of an element or compoun
Ways to express N: Number of replaceable ions (OH- ) =1
GEW = mol wt/no.of RI
1. N=Mxn = 40/1
2. M=mol/L  N=Mxn = 40gm
3. N=EW/L (where EW= mol wt/no.RI)
GEW for Salt
■ Valency is the # of shared positive or negative charges.
■ Equivalent Weight = Molecular weight of the Compound ÷ #EW=50g/49.038g ÷ 15L = 0.068N
Charge on the Compound.
________________________________________________
Prob: What is the normality of a solution that contains 1.5g of
Ca(OH)2 dissolved in 850ml?
WAYS OF EXPRESSING STRENGTH OF SOLUTION
ANSWER THE FF:
1. Calculate the normality of NaOH solution prepared
WAYS OF EXPRESSING STRENGTH OF SOLUTION by dissolving 2.916 g of NACl in water and then
topping it off with more water to a total volume of
■ Solve for 500 mL.
 Gram equivalent weight 2. What is the normality of a solution which was
prepared by dissolving 3.7 grams of Ca(OH)2 in
q.s. water to make 100 ml solution?
3. If 31.87 mL of base is required in the
standardization of 0.4258 g of KHP (meq wt =
0.20423), what is the normality of the base?
4. How many grams is needed to prepare 800 mL of
2M sodium chloride?
■ Equivalent weight depends on type of the substance. For 5. How many grams of NAOH will be needed to
example, prepare 2,000 ml 0f 0.15 M solution?
6. If 105 grams of NAHCO3 are dissolved in q.s. water
 Equivalent weight of acid = molecular weight of to make a 5,000 mL solution, what is the molarity of
acid/basicity of acid the above solution?
 The acid H2SO4 can donate 2 hydrogens. So, the 7. if you weigh 60 grams of NaCl and dissolved it in 1
basicity of H2SO4 is 2 Liter of solution: What is molarity, normality and
 Equivalent weight of H2SO4 = 98 /2 = 49 g/eq percent solution?
 Equivalent weight of salt = molecular weight of 8. compute for the Normality of a 2M Sulfuric acid.
salt/charge present in ionic form
■ What is the normality of a sulfuric acid solution that is 18
 The salt Na2CO3 ionizes to form 2Na+ and CO3 (–
M?
2).
 So, the charge present on both ions is 2. ■ Solution:
 Equivalent weight of Na2CO3 = 106 /2 = 53 g/eq
■ The molarity of a solution is defined as moles of solute per
 Equlivalent mass=molar mass/no. of cation liter of solution. The given concentration of sulfuric acid
Ex. Ways to Express N solution can be written as follows

I. Normality =M.n ■ Let's consider the ionization of sulfuric acid as follows

1. 0.15M HCl  ■ Hydrogen ion has an oxidation number of 1 and sulfate ion
2. 1.4M H2SO4  has an oxidation number of 2. The number of equivalence of
3. 2M NaOH  a solute is defined as the product of the number of positive
4. 0.4M Ba(OH)2  ions in a metal ion and the number of negative ions in a non-
metal ion. In this case, the number of equivalence of sulfuric
https://www.youtube.com/watch?v=QCZMyx_557I acid is 1 x 2 = 2.
Prob: What is the normality of a solution that contains 50g of ■ The normality of a solution is defined as the number of
H2SO4 dissolved in 15L? equivalence of a solute per liter of solution.
MW H2SO4= 98.076g
M=mol/L
N=M.  N=0.068N
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■ Therefore, the normality of a sulfuric acid solution is
or N=#EW/L