Pharmaceutical Analysis
KEY DEFINITIONS
Analytical Chemistry
• Analytical Chemistry: Branch of chemistry dealing with identification,
separation, quantification, and characterization of chemical components
in substances
• Pharmaceutical Analysis: Branch of analytical chemistry focusing on
evaluating pharmaceutical substances and products qualitatively and
quantitatively
Analysis Types
• Qualitative Analysis: Determines presence/absence of analytes
(detection and identification)
• Quantitative Analysis: Determines absolute or relative amounts of
analytes
• Structural Analysis: Establishes structure of samples/objects
Analytical Techniques
• Classical Analysis: Uses human senses, burettes, and balances
(titrimetry, gravimetry)
• Instrumental Analysis: Uses specialized instruments
(spectrophotometers, pH meters, mass spectrometers)
• Strong Electrolyte: Substance completely ionized in aqueous solution
• Weak Electrolyte: Substance only partially ionized in aqueous solution
Solution Chemistry
• Concentration: Amount of solute present in known amount of solution
• Molarity (M): Moles of solute per liter of solution
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• Molality (m): Moles of solute per kilogram of solvent
• Normality (N): Gram equivalents of solute per liter of solution
• Equivalent Weight: Mass that supplies/reacts with 1 mole of H+, OH-,
or electrons
pH & Buffers
• pH: Negative logarithm of hydrogen ion concentration
• Buffer: Solution containing weak acid/base and its conjugate that resists
pH changes
• Buffer Capacity (β): Amount of acid/base required to change pH of 1L
buffer by 1 unit
• Leveling Effect: Strong protophilic/protogenic solvents make all
acids/bases appear equally strong
• Amphiprotic: Substances that can both donate and accept protons (like
water)
Titration Terms
• Titration: Process of adding standard solution until reaction is complete
• Titrant: Standard solution added from burette
• Titrand: Substance being analyzed
• Equivalence Point: Point where titrant is chemically equivalent to
analyte
• End Point: Observable physical change indicating titration completion
• Indicator: Species that gives observable change at/near equivalence
point
• Primary Standard: Extremely pure, stable substance with high
molecular weight
• Secondary Standard: Solution whose concentration is determined by
titrating against primary standard
• Standardization: Process of determining exact concentration using
primary standard
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Non-Aqueous Titration
• Non-Aqueous Titration: Titrations carried out in absence of water for
very weak acids/bases
• Protophilic Solvents: High affinity for protons (enhance weak acids)
• Protogenic Solvents: Readily donate protons (enhance weak bases)
• Aprotic Solvents: Chemically neutral, don't undergo acid-base reactions
• Amphiprotic Solvents: Can both donate and accept protons
• Differentiating Solvents: Allow distinction between acids/bases of
different strengths
Salt Hydrolysis
• Hydrolysis: Reaction between ion and water producing H3O+ or OH-
• Salt of Strong Acid + Strong Base: No hydrolysis, pH = 7
• Salt of Weak Acid + Strong Base: Anion hydrolyzes, pH > 7
• Salt of Strong Acid + Weak Base: Cation hydrolyzes, pH < 7
CONCENTRATION UNITS & FORMULAS
Basic Concentration Units
Unit Formula Used For
% w/w (weight of solute / weight of solution) x 100 Ointments, creams
% v/v (volume of solute / volume of solution) x 100 IV fluids, injectables
% w/v (weight of solute / volume of solution) x 100 Liquid medications
ppm (mass of solute / mass of solution) x 10^6 Heavy metals, impurities
ppb (mass of solute / mass of solution) x 10^9 Ultra-trace contaminants
Mole-Based Concentrations
• Molarity (M): moles of solute / L of solution
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• Molality (m): moles of solute / kg of solvent
• Normality (N): gram equivalents / L of solution
• Mole Fraction (X): moles of component / total moles
Key Relationships
• N = M x n-factor
• Moles = weight (g) / molecular weight
• Dilution: C1V1 = C2V2
• 1 ppm = 0.0001% w/w = 1 mg/L
pH & BUFFER CALCULATIONS
Fundamental Equations
• pH = -log[H3O+]
• pOH = -log[OH-]
• Water: Kw = [H3O+][OH-] = 1.0 x 10^-14
• At 25°C: pH + pOH = 14
Strong Acids/Bases
• Strong acid: [H3O+] = molarity of acid
• Strong base: [OH-] = molarity of base x n(OH-)
Weak Acids/Bases
• Weak acid pH = 1/2(pKa - log C)
• Weak base pH = pKw - 1/2(pKb - log C)
• Relationship: pKa + pKb = 14
Henderson-Hasselbalch Equation
pH = pKa + log([conjugate base]/[acid])
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Salt Hydrolysis
1. Strong acid + Strong base → pH = 7
2. Strong base + Weak acid → pH = 1/2(pKw + pKa + log C)
3. Weak base + Strong acid → pH = 1/2(pKa - log C)
4. Weak base + Weak acid → pH = 1/2(pKw + pKa - pKb)
Buffer Capacity
β = moles added / ΔpH
Drug Ionization
• Acidic drugs: % ionized = 10^(pH-pKa) / (1 + 10^(pH-pKa)) x 100
• Basic drugs: % ionized = 10^(pKa-pH) / (1 + 10^(pKa-pH)) x 100
TITRATION FORMULAS & CALCULATIONS
Basic Titration Equation
(Va x Ma)/a = (Vb x Mb)/b
Where: V = volume, M = molarity, a,b = stoichiometric coefficients
Content Percent
Content % = (Weight of analyte found / Total weight of sample) x 100
Titer Value
Titer = (MWt x Concentration x Ratio_d) / (Ratio_t x 1000)
Factor
Factor = Theoretical concentration / Actual concentration
Titration Methods
1. Direct: Content % = (titer x volume x factor x 100) / weight
2. Back: Volume = V1 - V2
3. With blank: Volume = Sample reading - Blank reading
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Temperature Correction
Vc = V[1 + 0.001(t1 + t2)]
NON-AQUEOUS TITRATION
Common Solvents
• Glacial acetic acid (most common)
• Acetonitrile (with other solvents)
• Dioxane (non-leveling)
• DMF (protophilic)
Preparation of 0.1 N Perchloric Acid
• Mix 8.5 mL HClO4 + 900 mL glacial acetic acid + 30 mL acetic
anhydride
• Make up to 1 L, stand 24 hours
• Reaction: H2O + (CH3CO)2O → 2CH3COOH
Key Indicators
Indicator Color Change Use
Crystal violet Violet → Blue-green Adrenaline, Ergometrine
Oracet Blue B Blue → Pink Levodopa, Salbutamol
α-Naphtholbenzein Blue → Orange Bisacodyl, Metronidazole
Quinaldine Red Magenta → Colorless Pyrimethamine
Brilliant green - Metronidazole benzoate
Thymol blue - Fluorouracil (with Bu4NOH)
Azo violet - Bendrofluazide (with NaOCH3)
Thymolphthalein - Nalidixic Acid (with LiOCH3)
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IMPORTANT DRUG EXAMPLES
Non-Aqueous Titration Examples
Basic Drugs (with 0.1 N HClO4):
• Adrenaline, Metronidazole, Diazepam, Pyrimethamine
• Ephedrine HCl, Quinidine sulfate, Salbutamol sulfate
Acidic Drugs (with Bu4NOH or NaOCH3):
• Sulfonamides, Thiazide diuretics, Ethosuximide
• Chlorthalidone, Hydrochlorothiazide
Specific Calculations
• Diloxanide Furoate: 1 mL 0.1 N Bu4NOH = 0.03282 g
• Fluorouracil: 1 mL 0.1 N Bu4NOH = 0.01301 g
• Acetazolamide: 1 mL 0.1 N NaOCH3 = 0.02222 g
ANALYTICAL CLASSIFICATIONS
By Purpose
1. Qualitative: Identify presence/absence
2. Quantitative: Determine amounts
3. Structural: Establish structure
By Technique
1. Classical: Gravimetry, titrimetry
2. Instrumental: Spectroscopy, chromatography
3. Separation: Chromatographic methods
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KEY CONSTANTS TO MEMORIZE
• Kw = 1.0 x 10^-14 (at 25°C)
• Avogadro's number = 6.022 x 10^23
• Acetic acid Ka = 1.8 x 10^-5 (pKa = 4.74)
• Ammonia Kb = 1.8 x 10^-5 (pKb = 4.74)
• Blood pH = 7.4
PRACTICAL TIPS
Buffer Rules
• Buffer works best at pH = pKa ± 1
• Maximum capacity when [acid] = [base]
• Common biological buffers: bicarbonate, phosphate, proteins
Titration Rules
• All glassware must be completely dry for non-aqueous
• Use appropriate indicators for pH range
• Temperature affects volume measurements
• Primary standards must be pure, stable, high MW
Drug Ionization Rules
• pH = pKa: 50% ionized
• pH = pKa + 1: 90% ionized (acids)
• pH = pKa - 1: 90% ionized (bases)
QUICK REFERENCE FORMULAS
Concentration: C = amount solute / amount solution
Dilution: C1V1 = C2V2 pH: pH = -log[H+]
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Buffer: pH = pKa + log([A-]/[HA])
Titration: n1 = n2 (at equivalence point)
Ionization: α = √(Ka/C) for weak acids
Temperature correction: V2 = V1[1 + β(T2-T1)]
