Pharmaceutical Analysis

Quantitative

• Involves the test for the nature of the constituent of given material

• Comparison determination

• Ascertainment of impurities present in a given sample

• Answers the question: What is present in a given sample

Qualitative

• Determination of the proportion of components in a given sample

• Answers the question: How much is present in the sample?

1. Based on the extent of determination/ analysis

2. Based on the sample size/amount

3. Based on the nature of methods

4. Based on materials used

Classification of Analysis

Stretching

Distance between two atoms increases or decreases.

Symmetrical stretching

Move in the same direction

Asymmetrical stretching

Moves in opposite directions

Bending or Deformation

Position of the atom changes relative to the original bond axis. Generally, require less energy and take place at a longer wavelength than the corresponding stretching vibrations

Scissoring - close and open

Rocking - forward and backward

In-plane

Wagging - alternate from big to small

Twisting - the other mol is small and other is large

Out-of-plane

Voltammetry

Information about the analyte is derived from the measurement of current

Polarography

It is a method of analysis based on the measurement of current resulting from the electrolysis of an electroactive species at a given electrode potential under controlled conditions

Potentiometry

• Study and measurement of electrode potentials

• For pH determination (uses glass electrodes)

Indicating electrodes

Develops a potential that varies according to the activity (concentration) of hydrogen ions of the specific component in the solution

Reference electrodes

Maintains a constant potential

Chromatographic method

A process in which a solution of mixture containing inert materials, drug principles and impurities is separated into its components while moving through a bed of fixed porous solid having different and reversible affinities for the substance being separated

1 inch

Standard spotting at end of TLC plate

Stationary phase

Phase of chromatography having a fixed bed core pf large surface area

Silica gel G or Gypsum (Polar)

Adsorbent of stationary phase

Mobile phase

A phase which a fluid moves through or over the surface or stationary phase

Column Chromatography (Column, Mobile phase, Developer)

Paper Chromatography

Thin layer Chromatography

Classification of chromatography based on techniques

Column Chromatography (Column, Mobile phase, Developer)

The simplest type of chromatographic column consists of a separation flask and a cylindrical glass tube constricted at one end

• Mobile phase: Organic solvent

• Stationary phase: Silica gel

Paper Chromatography

Cellulose of the filter paper is sued as adsorbent

• Mobile phase: Organic solvent

• Stationary phase: Water

Thin Layer Chromatography

Separation of sample happens for only 30 minutes and utilize only 20 mg of sample

• Most distinct result and most precise

• Mobile phase: Organic solvent

• Stationary phase: Silica gel

Geiger Mueller Counter

Used to determine the position on the column of the radioactive compounds

Nernt’s Law

When 2 practically immiscible solvents are in contact with each other and a substance that is soluble in each is added, the substance distributes itself in such a way that at equilibrium and at a given temperature the ratio of the concentrations of the two solutions is constant

Limulus amoebocyte lysate

Reagent used in bacterial endotoxin test where if it is added in pyrogen it forms turbidity and gel-clot

Soxhlet apparatus

• a device that allows the continuous treatment of a sample with a solvent over a period of hours, or days to extract compounds of interests.

• only required where the desired compound has a limited solubility in a solvent, and the impurity is insoluble in that solvent

• used to extract alcohol soluble extractives in Benzoin

Digitalis

Pigeon

Insulin

Tubocurarine injection

Metocurine injection

Rabbit (HPLC)

Glucagon

Cat

Oxytocin

Chicken

Vasopressin

Male rat

Corticotropin

Rat

HCG

Chorionic gonadotrophin) - 20-23 days old

Female rats

Heparin

Protamine SO4

Sheep blood plasma

COD Liver oil

Rachitic rat (Spectrophotometer)

Parathyroid hormone

Dogs

In-vitro

In-vivo

Biological Reactivity Tests

In vitro

• determines suitability of plastic material intended for use as container of drug products

• quantifies the cytotoxicity of plastic materials and polymers

• includes agar diffusion and elution tests

Systemic injection test

Albino mice

Elastomeric material (Plastics)

Albino mice

Animal used in systemic injection test

Intracutaneous test

Albino rabbits

Elastomeric material (Plastics)

Eye irritation test

Albino rabbits

Medical devices and ophthalmic containers

Implantation test

Rabbits

Material in direct contact with tissue

In-vivo

Determines classification of plastic materials intended for use as container of drug products

Injection test

Systemic biological responses to plastic and other polymer

Intracutaneous test

Local biological response to plastic and other polymer

Implantation test

Reaction of living tissue to plastic or another polymer

Diethylthiocarbamate

Reagent used for limit test of Arsenic

Silver nitrate - AgNO3

Reagent used for limit test of Chloride

Barium chloride

Reagent used for limit test of Sulfate

Hydrogen sulfide TS = Black

Except:

• Zinc - white

• Cadmium - yellow

• Tin - orange

• Manganese- pink

Reagent used for limit test of heavy metals

Ammonium thiocyanate - NH4SCN

• Black-red complex

Reagent used for limit test of Iron

Ammonium cyanide

Ammonium citrate (violet color)

Hydroxylamine HCl

Dithizone

Reagent used for limit test of lead

Spectrophotometry (535, 545 nm)

Method used for Arsenic

Flame photometry

Method used for Calcium, Sodium, Chloride

Vitamin A

Vitamin K (635 nm)

Vitamin B12 (361nm)

Vitamin B3 (450 nm)

Steroids (525 nm)

Assay methods of Vitamins that uses Spectrophotometry

Vitamin D

Vitamin B6

Vitamin B9

Penicillin

Cephalosporin

Assay methods of drugs that uses HPLC

Barbiturates

Vitamin E

Assay methods of drugs that uses GC

Vitamin B1

Vitamin B2

Assay methods of drugs that uses Fluoremetry

Vitamin B3

Vitamin B5

Vitamin B12

Assay methods of drugs that uses L. plantarum

Insulin activity

Assay methods of drugs that uses Rabbit Blood Sugar Method

Penicillin

Amikacin

Cycloserine

Kanamycin

Demeclocycline

Doxycycline

Tetracycline

Tobramycin

Antibiotics that assayed by Staphylococcus aureus

Neomycin

Streptomycin

Antibiotics that assayed by Klebsiella pneumoniae

Gramicidin

Antibiotics that assayed by Staphylococcus faecium

Ca panthothenate

Antibiotics that assayed by Lactobacillus plantarum

Candicidin

Antibiotics that assayed by Saccharomyces cerevisiae

Chloramphenicol

Antibiotics that assayed by E. coli

Cyanocobalamin

Antibiotics that assayed by Lactobacillus leichmanii

Wash bottles

Usually consist of rubber-stoppered, flat bottom flasks of 500-1000 mL capacity fitted with smoothly bent tubes and a jet made flexible by means of rubber joint

Policeman

A piece of soft rubber tubing fitted to the end of the glass rod. Used to loosen from the walls of the containing vessel adhering particles that are not removable by a stream of water from the wash bottle

Gooch crucible

A perforated bottom upon which is bedded a mat of asbestos, thus making it possible to collect, wash, dry, and weigh ppt. in the same crucible

Burette

Graduated glass tubes of uniform bore throughout the whole length, used in the measurement of the variable quantities of liquid

More accurate than graduated cylinder and pipette

Pipette

Graduated to deliver a standard temperature the volume indicated

Volumetric flask

Used to make up standard solutions to a given volume

Graduated cylinders

Used in making approximate measure of volume