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Lab # 1 King Saud University College of Pharmacy 454 PHL (lab) Pharmacology Department

BIOLASSAY Introduction Biological standardization (Bioassay) It is defined as the determination of the potency of chemical and biological agents such as drugs, hormones, vitamins, toxins, and antitoxin by means of biological indicators (in a live animals, isolated tissues or cell line). Biological indicators: 1- blood pressure 2- blood glucose 3- muscle contraction 4- inhibition of growth of microorganisms Application of bioassay in pharmacology 1- Determination of drugs potency. 2- Screening of new agents isolated from plants, animals or chemical labs and find their field of activities. 3- Determine the therapeutic advantage of one drug over anther treatments. 4- Determination of the pharmacological activities of a new drug. 5- Establishment of SAR. Comparison between Bioassay and Chemical assay 1- Bioassay is less precise, more time consuming and more expensive than chemical assay.
2- In bioassay, active constituent and its structure do not have to be known. 3- Bioassay is more sensitive than chemical assay (detect even small amount of the drug).

Screening of drugs
It means thorough investigation of substance for pharmacological activity and evaluation of this activity i.e. scanning and evaluation. The main purposes of screening are to determine whether the new substance are worthy for further attention and to indicate which among them have the most interesting pharmacological properties.

Types of screening
A- Simple screening It involves the use of one or two simple tests to find substances having a particular property. For example, a single test for conc. of glucose in blood can be used to screen compound for hypoglycemic activity. B- Blind screening

It provides clues to potential activities of new drugs and indicates the fields of activity if they exist. It also shows pharmacological inertness if it exists. The chief purposes of the blind screening are to demonstrate whether these new drugs are worthy of further attention and to indicate which among them have the most interesting pharmacological activities. C- Programmed screening It is used when a new drug of specific type is to be screened for some pharmacological effects. Examples are screening of certain drugs on the cardiovascular system, CNS, kidney, blood etc. It includes the use of quantitative assay of the most interesting compounds and their comparison with drugs known to be quite active representative members of their pharmacological class. It also provides indications of potential side effects.

Steps for biological screening of a drug

Neuropharmacological tests To detect sedative, hypnotic, tranquillizer, psychomotor stimulant, muscle relaxant, analgesic, antipyretic, and peripheral vasodilator activities. Mice are usually used here because they are small, easy to handle, economic, mammals and have long life span. Signs to be observed include consciousness, awareness, motor activity, pain response and excitability. Other responses should be evaluated like pupil size secretions, heart rate, respiration, and writhing. Cardiovascular tests To study the action of new drug on blood pressure and heart rate usually on anesthetized rat or cat. A group of 4 tests is performed after recoding normal blood pressure including; electrical stimulation of vagus for 2 seconds, injection of 5 mcg/kg of Ach, occlusion of both carotid arteries for 45 seconds and injection of 5 mcg/ml of epinephrine. The cat nictitating membrane It is an additional eye lid that composed of smooth muscle innervated by post-ganglionic sympathetic nerve fibers. It has alpha and beta subtype of receptors. Nictitating membrane is attached to a lever for recoding its contraction. By the aid of this test we can determine whether the new drug is ganglionic blocker, adrenergic neuron blocker or alpha receptors blocker. Tests on isolated organs like guinea pig ileum, intestine, rat uterus, aorta etc. This is usually carried out to illustrate the type of receptors the drug act on.

Factors that can affect the pharmacological response to drugs

They include the following: Species and strain of animals; differ in biochemical, functional and morphological features thus their response to the drug will vary quantitatively or qualitatively. Example, morphine in CNS depressant in rabbit and human and CNS stimulant in cats and dogs. Sex; difference in drug response among different sex is attributed to difference in the level of metabolizing enzymes. Age; the level of metabolizing enzymes varies with age and some enzymes may be lacked in newly born animal. Diseases; the hypersensitivity to catecholamine action on the CVS is increased in hyperthyroidism. Environmental; such as temperature, seasons, nutrition, light and isolation of animal. For example, the convulsive action of insulin in mice is greatly influenced by temperature.

King Saud University Practical Pharmacology College of Pharmacy (454) Pharmacology Department 1

PHL Lab #

Introduction (Biological screening)

Points to be discussed 1- Definition of bioassay 2- Discussion of applications of bioassays in pharmacology 3- Comparison between Bioassay and Chemical assay 4- Screening of drugs: types and steps of biological screening 5- Factors that affect the response to the drugs Lab work Training on handling and injection of animals. Report Format Title, date Aim of study Introduction: examples of drugs, MOA, receptors, side effects, uses, anatomy, physiology, etc. Material: Drugs used (conc. & doses in table) please title each table e.g. table 1. the conc. and dose of drugs used. Instruments: example physiograph, transducer, hotplate, funnels, needles etc. Animal used: mice, rat, frog etc. Physiological solution, temperature, aeration gas (Only if the experiment involves isolated tissues). Method. Results (graphs, curves, equations, or tables) . Comment