hospital pharmacy lab manual

7/22/2019 Hospital Pharmacy Lab Manual

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LAB MANUAL OF HOSPITAL PHARMACY FOR PTU STUDENTS

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Experiment 1

To study the hot air oven and study its different parts

Reference:Nand Pratibha, Khar Roop K A text book of hospital and clinical

pharmacy Theory and Practical,6th edition, Birla publications(p) Ltd Delhi, pageno:473.

Theory: sterilization can be defined as a pharmaceutical process in which almost

99.9% of micro-organisms are removed from the Pharmaceutical products or

equipments by the application of a single or multiple techniques. An article would

be deemed sterile only when there is complete absence of viable micro-organisms

from it. The methods for sterilization can be divided in to three main categories.

I. Physical methods

Dry hear sterilization, moist heat sterization

Radiation sterilization

II. Chemical methods:

Sterilization by heating with bactericide

Gaseous sterilization

III Mechanical methods

Ceramic filter, Sintered glass filter Membrane filter

Hot air Oven (Dry heat sterilization): This process is suitable for heat-stable,

non-aqueous products, powders as well glassware and metal equipments. A

preparation to be sterilized by dry heat is distributed in the final containers which

are then either finally sealed or temporarily closed to exclude micro-organisms and

then heated so as to ensure that the entire contents of each container are maintained

for an effective combination of time and temperature to provide an adequate

assurance of sterility. Cycles of a minimum of 180 for not less than 30 minutes, a

minimum of 170 for not less than 1 hour or a minimum of 160 for not less than 2


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hours may be employed but other combinations of time and temperature may be

necessary for certain preparations. Containers that have been temporarily closed

during the sterilisation procedure are then finally sealed using aseptic technique so

as to exclude micro-organisms.

Dry heat sterilization is usually carried out in a sterilizing oven designed

specifically for the purpose. The oven is supplied with heated, filtered air,

distributed uniformly throughout the chamber and employing a blower system with

devices for sensing, monitoring, and controlling the critical parameters. A vent is

provided on both sides for the escape of vapors. A thermometer is provided in door

side to record the inside temperature of the chamber.

During dry heat sterilization the micro-organisms and bacterial spores are killed

by oxidation. Since dry heat is less effective than moist heat, hence higher

temperature and longer period is required.

Applications: Material/equipment(s) which is sterilized by dry heat include: Fixed

oils, glycerine, liquid paraffin, propylene glycol, glasswares, powders, surgical

instruments etc.

Disadvantages:

1. It requires long heating, high temperature and long exposure, unsuitable for

thermolabile substances.

2. Moist medicaments, rubber and plastic articles are destroyed by this method.

3. Preparations containing water, alcohol or other volatile substances cannot be

sterilized by this method.

4. It is unsuitable for surgical dressings

Experiment 2


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To study the Autoclave and study its different parts


pharmacy Theory and Practical,6th

edition, Birla publications(p) Ltd Delhi, pageno:478.

Theory: This process involving heating in an autoclave with saturated steam under

pressure should be used whenever possible for aqueous preparations and for

surgical materials. An aqueous product which is to be sterilized by this method is

distributed into suitable containers which are then sealed so as to exclude micro-

organisms. It is than exposed to saturated steam for a time sufficient to ensure that

the entire contents of each container are maintained for an effective combination of

time and temperature to ensure sterility.

The following combinations of temperature and time are normally employed.

Holding

temperature (C)

Minimum

holding time

(minutes)

115 to 118 30

121 to 124 15

126 to 129 10

134 to 138 3

In this method micro-organisms are killed due to coagulation or denaturation ofproteins in living cell of micro-organisms. It is more effective than dry heat

because saturated steam has more penetrating power than dry heat. More over the

thermal capacity of steam is much higher than the thermal capacity of dry heat.

Working: autoclave is a strong metallic chamber usually made up of stainless

steel. The material to be sterilized is packed in a clean fabric cloth and is placed in


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perforated chamber of an autoclave. The water level is checked so that it should

not touch the bottom perforated metallic chamber. The lid is closed with the help

of nuts and bolts. Autoclave is switched on and water is allowed to boil. Remove

the trapped air. Steam vent is then closed and pressure is allowed to rise up to 15

lb/sq inch and at this pressure saturated steam has a temperature of about 121oC.The pressure is maintained for 30 min. the autoclave is switch off allowed to cool,

remove the steam and open it.

Applications:

This method is used to sterilize bulk solutions, glassware, surgical dressings,

rubber gloves, culture media and surgical instruments are efficiently sterilized by

this method. Solutions packed in sealed containers such as SVP, LVP can be

sterilize by this method.

Disadvantages:

1. It requires high temperature and moisture, unsuitable for thermolabile

substances.

2. This method cannot be used for oily injections, fats, powders, ointments

because steam cannot penetrate them.

3. Moist medicaments, rubber and plastic articles are destroyed by this method.

4. Preparations containing water, alcohol or other volatile substances cannot be

sterilized by this method.

5. It is unsuitable for surgical dressings


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Experiment 3

To sterilize conical flask by Dry heat sterilization



Requirement: absorbent cotton, brown paper, warmwater, alcohol, soap solution,

chromic acid, hot air oven and conical flask.

Theory: This process is suitable for heat-stable, non-aqueous products, powders as

well glassware and metal equipments. A preparation to be sterilized by dry heat isdistributed in the final containers which are then either finally sealed or

temporarily closed to exclude micro-organisms and then heated so as to ensure that

the entire contents of each container are maintained for an effective combination of

time and temperature to provide an adequate assurance of sterility. Cycles of a

minimum of 180 for not less than 30 minutes, a minimum of 170 for not less than

1 hour or a minimum of 160 for not less than 2 hours may be employed but other

combinations of time and temperature may be necessary for certain preparations.

Containers that have been temporarily closed during the sterilization procedure are

then finally sealed using aseptic technique so as to exclude micro-organisms.

During dry heat sterilization the micro-organisms and bacterial spores are killed by

oxidation. Since dry heat is less effective than moist heat, hence higher

temperature and longer period is required.


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Procedure:

I) Preparation of articles before loading in hot air oven:

a. Conical flask should be thoroughly washed in following order

Acid wash (chromic acid), detergent wash, rinse with distilled

water followed by rinsing with alcohol.

b. Dry the container at 65oC

c. The flask should be stopped with the help of cotton and a piece of

paper and with a thread.

II: Loading of articles in hot air oven:

a. The container should be placed inside the hot air oven.

b. Close the door and switch it on

c. Adjust the temperature at 180oC, wait for the temperature to rise up

to 180oC. Maintain the temperature for 30 minutes.

d. Switch off the source and wait for 15 to 20 minutes until the

temperature of hot air oven fell down below 60

o

C.e. Open the hot air oven, unload the materials and transfer to aseptic

room for further processing.

Conclusion: The Conical flask was sterilized and submitted.

Experiment 4


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To prepare 5%w/w dextrose solution and sterilize by autoclave




Requirement: Dextrose, distilled water, Conical flask, autoclave.

Theory: In this method micro-organisms are killed due to coagulation or

denaturation of proteins in living cell of micro-organisms. It is more effective than

dry heat because saturated steam has more penetrating power than dry heat. More

over the thermal capacity of steam is much higher than the thermal capacity of dry

heat. The standard operating conditions are at 121oC for 15 minutes at 15 lb/sq

inch.

Procedure:

I) Preparation of Material for loading in autoclave

First weight 2.5 gm dextrose and make volume up to 50ml with water in conical

flask. Then flask used is plugged with non-absorbent cotton wool, and then it is

covered with brown paper, held by string.

II) Loading inside an autoclave:The sufficient water was placed inside an autoclave and place the tray

just above it. Close the lid of autoclave, switched on and water is allowed

to boil. Remove the trapped air. Steam vent is then closed and pressure is

allowed to rise up to 15 lb/sq inch and at this pressure saturated steam has

a temperature of about 121oC. The pressure is maintained for 30 min. the

autoclave is switch off allowed to cool, remove the steam and open it.

Conclusion: The Conical flask was sterilized and submitted.

Experiment 5


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To carry out filling and sealing of given ampoules

Reference: Remingtons The Science & Practice of Pharmacy, Mack Publishing

Co. Easton, PA

Requirements: Sodium chloride, distilled water, syringe, methylene blue, beaker,

Bunsen burner, forceps, ampoules.

Theory: Glass Ampoules are primary containers used packaging of single dose

small volume parenterals. The product is filled in to ampoules with semi-automatic

or automatic machines under aseptic conditions. The materials to be used must be

sterile. The filling of materials can be done with hypodermic syringe attached with

long needle. While filling in to ampoules, precautions should be taken that needledont touch to the site of ampoules to prevent cracking and staining at the time of

sealing. The filling and sealing operation should be carried out under aspetic

conditions. The sealing of ampoules can be done by two methods

1. Tip sealing

2. Pull sealing method.

Procedure:

1. Prepare sodium chloride solution by dissolving 0.9 gm of sodium chloride in

100 ml of distilled water, filter it.

2. Transfer the sodium chloride solution in to ampoules with hypodermic

syringe.

3. Ampoule tip was melted using Bunsen burner with continuous rotation of

the ampoule.

4. When the glass was melted, the tip of the ampoule was stretched by using

forcep.

5. Now the sealing of ampoule completed.


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6. Put the sealed ampoules in methylene blue solution to detect the leakage.

7. Take out the ampoules from the methylene blue solution, wash the external

surface. And observe under light.

8. Color of the solution in the ampoules should remain unchanged. If blue color

observed, discard the ampoule.

9. Label the ampoule and stored for sterilization.

Result: Filled, sealed and the labeled ampoules are submitted to laboratory.

Experiment 6

To prepare Calcium Gluconate I.V Injection


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Requirements: Distillation still, autoclave, ampoules, syringe, Bunsen burner,

forceps.

Theory: This article is water purified by distillation or by reverse osmosis and it

meets the purity requirements stated under Purified Water (see IP monograph). It is

not intended to be sterile but should comply with the tests for pyrogens, or with the

test for a limit of bacterial endotoxins, it must be produced, stored and distributed

under conditions designed to prevent production of pyrogens or endotoxins.

The bacterial endotoxins test (BET) measures the concentration of bacterial

endotoxins that may be present in or on the sample of the article to which the test is

applied using a lysate derived from the hemolymph cells or amoebocytes from the

horseshoe crab,Limulus polyphemus. The Endotoxin limit for a given test material

of preparation is calculated from the expression K/M, where K is the maximum

number of Units of endotoxin which the patient may receive without sufferingtoxic reactions and M is the maximum dose administered to a patient per kg per

hour. The value of K is 5.0 EU/kg for injectable preparations except for those

administered intrathecally and is 0.2 EU/kg for intrathecal preparations. For this

calculation, it is assumed that the average person weights 70kg.

Procedure:

1. Glass distillation unit is used for preparing water for injection.


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2. The ampoules and glassware to be used should be treated with chromic acid,

followed by washing with purified water.

3. Switch on the distillation unit.

4. Reject the first part of the distillate.

5. Collect the water for injection in a receiver.

6. Transfer 2 ml of water for injection in to an ampoule with the help of

syringe.

7. Seal the ampoule immediately.

8. Sterilize with in 12 hr of production, label and submit.

Report: the water for injection was prepared, labeled and submitted to the

laboratory.

Experiment 8

To sterilize the rubber gloves by Moist Heat Sterilization


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Requirements: Autoclave, conical flask, cotton, rubber gloves.

Theory: Autoclave is a strong metallic chamber usually made up of stainless steel.

The material to be sterilized is packed in a clean fabric cloth and is placed in

perforated chamber of an autoclave. The water level is checked so that it should

not touch the bottom perforated metallic chamber. The lid is closed with the help

of nuts and bolts. Autoclave is switched on and water is allowed to boil. Remove

the trapped air. Steam vent is then closed and pressure is allowed to rise up to 15

lb/sq inch and at this pressure saturated steam has a temperature of about 121oC.

The pressure is maintained for 30 min. The autoclave is switch off allowed to cool,

remove the steam and open it.

Procedure:

1. The rubber gloves require packing in such a way that it should allow the

steam penetration inside the fingers and all the surface of the gloves.2. The gloves are lubricated with talcum powder or starch powder before

sterilization.

3. The wrist are turned back so that the outside need not be handled when the

gloves are put on

4. Close the lid of the autoclave, Remove the trapped air. Steam vent is then

closed and pressure is allowed to rise up to 15 lb/sq inch and at this pressure

saturated steam has a temperature of about 121oC. The pressure is

maintained for 30 min. the autoclave is switch off allowed to cool, removethe steam and open it.

Precautions:


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1. Dont open the lid immediately.

2. While loading the autoclave, each article should be placed in such a way that

it should be exposed to the steam uniformly.

3. The complete wrapping of the material is essential to avoid the

contamination by non-sterile factors.

Report: The rubber gloves were sterilized by moist heat sterilization.

Experiment 9

To Evaluate the Given Prescription





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Theory: Prescription drug order is defined as a lawful order from a practitioner for

a drug or device for a specific patient, including orders derived for collaborated

Pharmacy practice that is communicated directly to a pharmacist, in a licensedpharmacy.

The terms prescription drug order, prescription order and prescription are one and

the same used interchangeably by health care workers and the public.

Prescriptions are most commonly used to describe drug orders for ambulatory

patients (out patients) who get their prescribed medications from retail pharmacies.

The term medication order is usually used when referring to drug orders for

persons who are patients in hospitals. These patients are referred to as in patients.

Different parts of Prescription:

1. Date

2. Name, age, gender and address of the patient

3. Superscription

4. Inscription

5. Subscription

6. Signatura7. Renewal instructions

8. Signature, address and registration number of the Physician

HOSHIARPUR MEDICAL CENTRE AL-MADINA STREET

HOSHIARPUR, PUNJAB Rx 1120

Name: SURINDER Date : 00/00/00


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Report: The given prescription was evaluated.

AddressL 110, MAIN STREET,

CENTRAL TOWN, HOSHIARPUR

Age: 40 yr.

Wt:

Ht:

Rx

PHENOBARBITONE TABLET 10 mg

ALUMINIUM HYDROXIDE TABLETS

# 30

Sig: 1 Tab Each t. i .d . for 10 days

Refi l l : 0

Dr. N.K SHARMA, MD

RE G N o. _ _ __ _ __


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Experiment 10

To Prepare Distilled Water

Reference:Nand Pratibha, Khar Roop K A text book of hospital and clinicalpharmacy Theory and Practical,6th edition, Birla publications(p) Ltd Delhi, page

no:485.

Theory:

Distillation can be defined as a process or unit operation in which the separation

of two different liquids or the separation of impurities from a liquid can be done. It

is based on the boiling point and the volatility of the components present in the

mixture. When two liquids are mixed together, they may be miscible with each

other, such type of liquid mixture is known as binary mixture of liquid

Example: ethanol and water

Liquid boils when its vapor pressure is equal to atmospheric vapor pressure.

Simple distillation is conducted at its boiling point of the liquid. So higher the

volatility of liquid, better is the separation by simple distillation.

Applications of distillation:

1. Simple distillation method is used for preparation of distilled water and this

can be used further for preparation of water for injection.

2. Volatile and aromatic water are prepared.

3. Organic solvents are purified

4. Non volatile solids are separated from the volatile liquids.

Procedure:

The normal tap water was introduced in to the distillation flask, was heated to the

boiling point.


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The vapor is cooled, condensed by the condenser and is collected.

Precautions:

1. The water should be filled up to 2/3rd

volume of the flask.

2. The porcelain bits may be added as antibumping agents.

3. First some part of the distillate should be discarded.

Report: Distilled water 50 ml was prepared and submitted.


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Experiment 11

To find out the percentage variation in length and width of the given sample

of bandages.

Reference:Nand Pratibha, Khar Roop K A text book of hospital and clinicalpharmacy Theory and Practical,6

thedition, Birla publications(p) Ltd Delhi, page

no:490.

Requirement: Roll of bandage, Meter scale, Thin wire

Theory: It is a strip of plain woven cloth for wrapping and binding body parts. It

is available in continuous length which is used along with medicaments on

wounded surface. Fabric is woven from the staples of bleached white cotton havingcotton having no taste or odor. Cloth is free from defects of weaving.

Procedure:

Length: length of bandage is measured after equilibrating the sample in a humidity

chamber at 65 20% humidity and 27 2oC temperature for 24 hours or more. The

sample of bandage is unrolled and stretched from one end on wards. The sample of

the bandage is unrolled and stretched from one end onwards. The length is

measured from one end to other end with meter scale avoiding any wrinkles or

creases. Percentage of length is calculated from the following formula.

A = % of length = M/D x100

M= Measured length of bandage

D = Declared length of bandage on label.

The variation in length can be calculated as:

V = A-100

Where A = % of declared length available. The variation is indicated by the sign +

or -.

The variation in the width is measured in the same manner.


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Experiment 12

To perform the identification test for absorbent cotton wool



Requirements: Test tube, Test tube holder,Iodine solution, sulphuric acid

hydrochloric acid, ammonical copper oxide solution, formic acid Phenol, acetone.

Identification test

Ignition test:

1. Ignition test can be done by two methods.a. By advancing the fibre towards the flame.

b. By heating in a porcelain dish.

c. Cotton burns with a flame and gives very bitter odour or fumes.

2. Cotton fibres are moistened with N/50 iodine and when dry, 80% w/w.

sulphuric acid is added, blue color is produced.

3. Absorbent cotton wool dissolve completely with uniform swelling in

ammonical copper oxide solution.4. In sulphuric acid 80% w/w solution, it dissolves.

5. In cold sulphuric acid 60% w/w solution is insoluble.

6. In warm Hcl, it is insoluble.

7. It is insoluble in formic acid and phenol.

8. It is insoluble in acetone.

hospital pharmacy lab manual

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