Pharmaceutical Analysis 2: Instrumental Methods of Analysis

(Review of Analysis 1 and Introduction)

Qualitative- checking for the presence or absence of a component (e.g. identification testing)
Quantitative- purity of the compound; determination of the exact amount of the specific compound (e.g. percentage purity)

Type of Analysis:
1. Based on the amount of sample to be analysed:
a. Ultra micro- < 1.0mg
b. Micro- 1.0-10.0mg
c. Semi micro- 10.0-100.0mg
d. Macro- 100.0-1,000.0mg/1.0g
2. Based on the extent of analysis
a. Proximate- analysing a specific group of compound (e.g. total alkaloidal content of coffee bean: xanthine alkaloid)
b. Ultimate- analysing a specific single compound (e.g. determination of total “caffeine” content of coffee bean)
3. Based on the nature of method
a. Chemical -aka “General Method” “Wet Method” “Classical Method”
-usually used, but is often subjected to bias
-Examples: Titrimetry Method; Gravimetry Method
b. Instrumentation –method employs the use of specific instruments
-Examples: Chromatography, Mass Spectrometry, UV-Vis Spectroscopy
c. Special Methods -usually employed in chemicals of botanical origin
-Examples: Ash content; Constant for Fats and/ Esters; Saponification Value
d. Materials used -according to the materials being used in the conduct of the analysis
 Chemical Testing- used of chemical reagent
 Physical Testing- used of index of purity (e.g. BP, MP, Optical Activity)
 Biological Testing- animal assays/models; Microbiological assays (e.g. Assays of ABX)
Chemical/ General Method
I. Titrimetry
Definition of terms:
Titrant -aka “standard solution” “volumetric solution” “solution with known concentration”
Titrand -aka “analyte” “sample”
Indicators -the substance that helps visualize the end points
-compound that is capable changing color near or at the endpoint
End point -actual or experimental point at which equal amount of the analyte and titrant had been reacted
-the point where physical change usually occurs (e.g. change of color)
Equivalence point –theoretical point at which equal amount of analyte and titrant had been reacted (not the actual)
Precision -the closeness of two or more actual value
Accuracy -the closeness of an actual value to the theoretical value (basis of Absolute Error)
Errors -the discrepancy between a computed, observed or measured value or condition and the true, specified,
theoretically correct value or condition. Types of Error:
a. Random –aka “indeterminate error” (affects precision)
-cause: unknown; maybe due to chance
b. Systematic -aka “determinate error” (affects accuracy)
-cause: identifiable; maybe due to wrong process of analysis/ adulterated chemicals
Standardization –process of determining the exact concentration of a solution (comparing a solution to a standard)
a. Primary standard –substance of high degree of purity, used in direct standardization purposes
b. Secondary standard –solution of known concentration used in indirect standardization purposes
Titer -amount of substance (in milligrams) chemically equivalent to 1mL of the standard solution
Blank determination –the process of repeating the entire procedure but omitting the sample or analyte in the process
Purpose:
a. Base reacts to glass, thus certain test are involved in manufacturing process
b. For correction
c. To enhance reliability of the endpoint
Classifications:
1. Base on the process of titration
A. Direct Titration –one titrant is involved in the process
B. Residual Titration –aka “Blank Titration”; two titrants are involved in the process
Importance:
 When sample is insoluble (sample reacts when it is in water)
 If the sample is volatile (loss of sample due to evaporation)
 If reaction is too slow
 If it does not give a sharp endpoint (thus 1st VS is added in excess and 2nd VS is used to titrate the excess of the 1st VS)

2. Base on the Reactions involved

A. Neutralization: Acid-Base Titration
 Aqueous- Solvent is water
Aqueous Indicators used:
SA + SB= Phenolphthalein (Basic endpoint-pink; Acidic endpoint-colorless)/Methyl Red (Basic endpoint-orange; Acidic endpoint-red)
WA + SB= Phenolphthalein (Basic endpoint)
WB + SA= Methyl Red (Acidic endpoint)
WA + WB= no reaction
 Non-aqueous- Solvent is nonpolar/non-aqueous
Non-aqueous Indicators:
-for very weak acids and bases
-Examples: malachite green, gentian violet, quinaldine red, methylrosaniline Cl, azoviolet, thymol blue

a. Acidimetry- measurement of base by a standard acid; VS: Acid

b. Alkalimetry- measurement of acid by a standard base; VS: Base
c. REDOX –process of titration undergoes reduction/ oxidation reaction
d. Complexation –process involves the chelation process of the transition metal group
e. Precipitimetry –process commonly involves determination of the presence of halides through identification of precipitate