manual of laboratory experiments-old

8/13/2019 Manual of Laboratory Experiments-old

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AL-NAHRAIN UNIVERSITY

COLLEGE OF MEDICINE

DEPARTMENT OF PHARMACOLOGY

& THERAPEUTICS

MANUAL OF LABORATORY EXPERIMENTS

Prepared by

Dr. Ahmed Haqi Ismael

2009 –2010


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List of Contents

!"eToi#

$ Ho% to %ite ! '!(o!to) eot

*+- H!n,'in" of !ni!'s

./- Ro0ts of !,inist!tion

1$- T2e Resonse of 20!n s3in to Hist!ine & A,en!'ine

4*- Effe#t of P!!s)!t2oieti#s on "'!n,0'! se#etions

+5.- D0"s !#tin" on e)e

+$6- Effe#ts of D0"s on t2e Atei!' B'oo, Pess0e of H0!n

+.7- E8!'0!tion of se,!ti8e !n, to9i# effe#ts of A'#o2o's

+61- E8!'0!tion of Anti-inf'!!to) D0"s

+74- E8!'0!tion of An!'"esi#s

+1+5- Gene!' !nest2esi!

+4++- Anti#on80's!nts

/5+/- Antiei'eti#s

/++$- Dos!"e fo

/.+*- Pes#ition %itin"

2


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Ho% to %ite ! '!(o!to) eot

Each student should write his report by himself using his own words to

improve his language. DO NOT COPY. The report should be brief, precise and including the following items:

1- The name of the student and the experiment.

2- The principle of the experiment.

- The aim of the experiment.

!- "aterials and methods used.

#- $esults, including the measurements and observations during the

experiment that sometimes should be arranged in a form of table.

%- &iscussion and conclusion, which is the ost iot!nt item in the report

as it shows the ability of the student to discuss his findings and compare

them with those mentioned in the textboo'.


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E9: No: +

HANDLING OF LABORATORAY ANIMALS

The pharmacological experiments will be mostly done on laboratoryanimals (mice, rats, ) rabbits*, so the animals should be treated well, as

improper handling and treatment of animals may lead to failure to observe

the effects, and will give rise to bad results. +o we examine the animals as

following:

I- Gene!'

1- ealth: healthy or sic' animal

2- ctivity: hyperactive, irritable, aggressive (/+ stimulants e.g.strychnine* 0 hypoactive (e.g. sedatives* 0 normally: mouse is

hyperactive, rat is aggressive, and rabbit is uite.

- ait: ataxic or drun'en gait (e.g. alcohol* 0 normal.

!- Erection of tail: in rats due to stimulation of spinal cord by

narcotics (e.g. morphine* called straub phenomenon.

#- $ighting reflex: placing the animal on the lateral side, the animal

will retain its normal position. &ecreased by /+ depressants (e.g.

dia3epam* and increased by /+ stimulants (e.g. strychnine*

%- 4ain reflex: reflex to painful stimulation by twisting the patella in

rabbit or pinching the tail by forceps in rats ) mice. (&ecreased

by narcotics ) non-narcotics*.

II- Vit!' si"ns

Each reading to be ta'en several times by different persons,

ignore the odd one and ta'e the mean of remaining readings:

• $espiratory rate (abdominal breathing 0 nostrils*

• heart rate (flic'ering*

III- E9!in!tion of e)e

1- on5unctival blood vessels: - congested (dilated blood 6essels*

- pale (constricted blood 6essels*

2- 4upil si3e: - dilated (mydriasis* or constricted (miosis*

- 7ight reflex: by shading the eye for 8 seconds then switch the

light on and this will result in miosis.

!- orneal reflex: attaching cornea by piece of cotton wasp from the

lateral side resulting in eye blin'ing.

E9: Nee,s; Fo#es


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E9: No: /

Ro0tes of !,inist!tion

Definition $oute of administration is defined as the mean by which drugs can be

given into the body.

F!#tos to (e #onsi,ee, %2en #2oosin" #et!in o0te;1. Site of ,0" !#tion: e.g., treatment of ertain !I" diseases

neessitates gi#ing the dr$g orally.

2. D0" n!t0e% intra#eno$s fl$ids sho$ld be isotoni.

. Onset of !#tion: treatment of emergeny onditions neessitates

the $se of intra#eno$s ro$te.

!. D0!tion of !#tion: dr$gs intended for longer d$ration of ation

are gi#en by a ro$te &hen absorption is slo&.#. P!tient st!t0s: oral ro$te an not be $sed &hen the patient is

$nonsio$s, or has diffi$lty in s&allo&ing, or has repeated

#omiting.

%. Desie of t2e !tient.

Ro0tes of !,inist!tion #!n (e #'!ssifie, !s fo''o%s;

. Enteral (9T*: oral

. 4arenteral: intramuscular (9"*, intravenous (96*, subcutaneous

(+*, inhalational.

9n the beginning measure the weight of the animals to ensure the

precision of dosage of the given drugs by using measuring scale

(balance*.

+: O!' o0te:

y placing mouth gag into the mouth of rabbit, insert rubber tube

to about 28 cm. To ma'e sure that the tube is in the esophagus and not

in the trachea, dip the end of the tube into a bea'er containing water(bubbling indicates wrong position*. y using medical syringe, push

2.# ml of normal saline (/.+* into the stomach through the rubber

tube.

A,8!nt!"es:

• +imple, convenient, and acceptable.

• ;ral drugs can be given in different dosage forms.

• +afe route: since in overdose, it can be managed easily.

• The drug can be placed at the site of action (e.g., antihelminthics*.

Dis!,8!nt!"es:• +ome drugs are ,esto)e, in t2e "0t (e.g., some penicillins,

insulin, oxytocin*

#


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• Tablets ta'en with too small a uantity of liuid and in supine

position can lodge in the oesophagus with delayed absorption and

may even cause ulceration (e.g., doxycycline*.

• +ome drugs may cause "!sti# iit!tion.

• A(sotion may be affected by foo, or by ot2e ,0"s which

inhibit gut motility e.g., antimuscarinic and opiods.

• Fist !ss et!(o'is (by intestinal wall and by the liver* limits

the efficacy of some drugs when ta'en orally.

/: Int!8eno0s 8.82 ml* by insulin syringe.

&rugs should be given slowly. 4ress cotton onto the wound.

A,8!nt!"es;

• 7arge volumes can be given via this route.

• =nconscious patient.

• $apid onset of action.

• +uitable for continuous infusion (for rapidly destroyed

drugs*

• +uitable for too irritant drugs (anticancer drugs*.

• /o first pass metabolism.

Dis!,8!nt!"es;

• 96 fluid should be aueous, and (isotonic*

• 9nfection and local venous thrombosis.

• The in5ected drug can not be recalled simply in case of toxicity.

$: Int!0s#0'!


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• /ot acceptable for self administration

• "ay be painful

*: S0(#0t!neo0s


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E9: Nee,s; Mo0t2 "!"> NG t0(e> (e!3e> s)in"es ('!,e> 9)'o'> !'#o2o'> #!n0'!> 2e!in> '!ste>

nee,'es> #otton> ?!> et2e> N:S> (!'!n#e:

E9: No: $

T2e Resonse of 20!n s3in to Hist!ine

& A,en!'ine

Ai; To show the effect of istamine and drenaline on the human s'in.

Pin#i'e; istamine affects the human s'in in the term of Triple

response of 7ewis .This action can be antagoni3ed by

a - ntihistamines (e.g. diphenhydramine* by bloc'ing histamine

receptors in the s'in, so this is called competitive antagonism. b A drenaline by producing action that reverses the action of histamine,

so this is called physiological antagonism.

Met2o,;

Birst, cleanse the forearm with a swab of cotton wool soa'ed in alcohol

and leave it to dry. Then by using a lancet, ma'e four transverse parallel

pric's on the s'in of forearm separated from each other by distance of

five centimeters. The depth of pric'ing should be superficial i.e. the route

of administration is intra-dermal (9.&* namely such as not to draw blood.Then do the following steps:

a- 4lace few drops of istamine (1:1888* on the first bric'.

b- s a control, place few drops of normal saline on the second bric'.

c- ;n the third bric' ,place few drops of 1:1888 solution of

antihistamine as competitive antagonist ,then after 1-2 minutes add

few drops of histamine (1:1888* on the same site.

d- ;n the fourth bric', place few drops of adrenaline as physiological

antagonist.

e- ompare the results seen on each site.

N:B:+- To o(t!in (ette es0't> it is efe!('e to ,o t2e !(o8e

o#e,0e on esons %it2 f!i s3in:

/- T2is e9eient is not to (e ,one on !toi# in,i8i,0!'s:

E9: Nee,s; N:S> !nti-2ist!ine> !,en!'ine> 2ist!ine> Dos>

'!n#et> #otton> !'#o2o':

C


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E9: No: *

Effe#ts of P!!s)!t2oieti#s on

G'!n,0'! Se#etionsPin#i'e; The 4arasympathomimetic nervous system innervates a large number

of organs. The neurotransmitter acetylcholine (ch* mediates the

transmission of impulses from the preganglionic neurons to

postganglionic neurons as well as the transmission of impulses from

postganglionic nerve terminals to the effector organs. The action of ch

at the effector organ can be mimic'ed by drugs li'e arbachol,

methacholine, or muscarine. The sites at which ch and the

4arasympathomimetics act are called "uscarinic receptors (" receptors*and they are sensitive to bloc' by atropine. ch stimulates the secretion

of different glands in the body, li'e salivary or tear glands. 9n the rat,

there is a special gland called arderian gland. This gland is responsible

for the secretion of porphyrins, and is very sensitive to

4arasympathomimetics, which cause the secretion of bloody tears or what

is called chromodacryorrhesis.

P!!s)!t2eti# G!n"'ion Effe#to o"!n

↓ ↓ ↓ Pe"!n"'ioni# ost"!n"'ioni# ●‹ ●‹ ● ╣╠ ↑ ∟Ni#otini# e#: ↑∟M- e#:

A#2 A#2

Ai of e9eient;9s to show the stimulatory effect of 4arasympathomimetics on the

secretion of tears in the rat.

Met2o,s;1- 9n5ect 8.# mg0'g of arbachol 9.4 into a rat. Examine the eyes for

tears by wiping the eyelids with cotton to detect the bloody tears.

/ote salivation and nasal secretion.

2- 9n5ect another rat with 2mg0'g of atropine 9.4, wait about 1#-2#

minutes, and in5ect the rat with arbachol (8.# mg0'g* 9.4 and

examine for bloody tears, salivation and nasal secretion.

D


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E9eient!' nee,s; C!(!#2o'> !toine> s)in"es> nee,'es>#otton> !n, (!'!n#e:

E9: No: .

D0"s !#tin" on t2e e)e

The main compartments of the human eye (as shown in figure 1* are

cornea, iris, lens, ciliary body and vitreous humour.

Bigure 1: The composition of the human eyeIis; That involves:

ircular muscle ("uscarinic receptors*.

$adial muscle (lpha-receptors*.

Miosis; is due to either contraction of circular muscle or relaxation of

radial muscle.

M),i!sis; is due to either contraction of radial muscle or relaxation of

circular muscle.

lpha-agonist → ontraction of radial muscle of 9ris ("ydriasis*.Bear (+ympathetic discharge*.

&eath (7ac' of muscular tone due to lac' of ch.*

Except opiod intoxication ("-agonist 4in point "iosis*

lpha-loc'er $elaxation of radial muscles of 9ris ("iosis*

7ens: ttached to the ciliary body by ligaments (figure 2*.

18


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Bigure 2: +agittal section in the eye showing the lens and ciliary body

Ci'i!) (o,); that involves

- iliary epithelium (2 receptors*: responsible for secretion of aueous

humor.

- iliary muscle (" receptors*: responsible for near or far vision.

Bigure : The contraction and relaxation of the lens

11


12/26

Ci'i!) M0s#'e 0i' s#!'e> !n, #otton.

12


13/26

E9: No: 6

Effe#ts of D0"s on t2e Atei!' B'oo, Pess0e

Of H0!n

Ai;

1- To show the effect of some drugs on the arterial blood pressure.

2- To show the psychological effect of placebo drugs on the patient.

- To show the difference between single and double blind techniuesin the measurement of blood pressure.

Pin#i'e;

1- Ephedrine is a sympathomimetic agent i.e. it has presser effect

through stimulation of H-receptors (centrally*. ?here as,

chlorproma3ine has H-bloc'ing action through its effect on the

autonomic nervous system .+o it acts as antihypertensive.

2- 4lacebo drug is an inert substance that has no pharmacological action

(as lactose*.

M!tei!'s;

1- &rugs: ephedrine (1#mg*, chlorproma3ine (#8mg*, lactose powder.

2- pparatus: sphygmomanometer, stethoscope.

- Three volunteers to ta'e the drugs.

Met2o, s;

There are two methods to measure the blood pressure< firstly, thesingle blind techniue, where the volunteer does not 'now the nature

of the drug that he administered, where as the examiner (the person

that measures the blood pressure* 'nows that drug.

+econd method is called the double blind techniue, that involves

neither the volunteer, nor the examiner 'nows the nature of the drug

that administered.

The second method is more accurate than the first one because it

excludes the psychological effect of 'nowing the drug that affects the

measurements and the results.

1


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T2e ,o0('e ('in, te#2ni0e #!n (e !#2ie8e, !s fo''o%in";

1- The students are divided into groups, each group is consist of three

volunteers (to ta'e the three drugs*, an examiner (to chec' the

blood pressure*, and a registrar (to write down the measurements

for each volunteer*.

2- t the beginning (3ero time*, the examiner should chec' the blood

pressure of the three volunteers before administration of the drugs.This is considered as baseline .4. which should be noted by the

registrar of the group.

- The three drugs should be referred to as drug , drug , and drug

and not by their generic names.

!- /ow, each volunteer will administered a certain drug orally.

#- The blood pressure of the three volunteers should be chec'ed by

the examiner of the group every 18 minuets after ta'ing the drugs

and the measurements should be wrote down by the registrar of the

group in a form of table.%- "easuring the blood pressure in one group should be done by the

same examiner, using the same apparatus and by ta'ing three

successive readings each time.

Res0'ts; y comparing the measurements and the results obtained, you can

discover what is drug , , and . &iscuss that accordingly.

E9eient!' nee,s; E2e,ine> #2'oo!@ine> '!#tose>"e'!tin #!s0'es> stet2os#oes> !n, s2)"o!noetes:

1!


15/26

E9: No: 7

E8!'0!tion of se,!ti8e !n, to9i# effe#ts of A'#o2o's

Ai;

To measure the sedative and toxic effects of methyl and ethyl alcohols.

Pin#i'e;

oth methanol and ethanol are competing for the same en3yme for their

hepatic metabolism that is alcohol dehydrogenase.

This fact can be useful in the management of methanol poisoning:

Et2!no' ----------- A#et!',e2),e ---------------- A#et!te

A'#o2o' ,e2),o"en!se

Met2!no' -------- Fo!',e2),e --------------- Fo!te

s ethanol has higher affinity for alcohol dehydrogenase en3yme thanmethanol, so it is life saving in the emergency treatment after accidental

administration of methanol.

"ethanol causes blindness, acidosis, respiratory depression and death.

Met2o,s;1- measure the weight of two rats and chec' the following

parameters:G!it> !in ef'e9> esi!to) !te> onset of !nest2esi! !n, ,e!t2 if

o##0s:

2- 9n5ect one rat with methanol in a dose of mg0'g 9.4 and the other

rat with ethanol in the same dose.

- $ecord the time of administration and rechec' the above

parameters every # minutes to see the effects of both alcohols.

E9eient!' nee,s; Me!s0in" (!'!n#e> fo#es> s)in"es>nee,'es> et2!no'> !n, et2!no':

1#


16/26

E9: No: 1

E8!'0!tion of Anti-inf'!!to) D0"s

Ai;

To test the anti-inflammatory activity of acetyl salicylic acid.

Pin#i'e;

cetyl salicylic acid is the protype of non-steroidal anti-inflammatory

drugs (/+9&s*. These drugs can influence the process of inflammation

by bloc'ing the pathway of prostaglandins (4s* biosynthesis by

inhibiting conversion of rachidonic acid to precursor cyclic

endoperoxides by the en3yme cyclooxygenase (;G*.

COX-+>COX-/ .-Lio9)"en!se

↓ ↓ PGG/ ←--------- A!#2i,oni# !#i, ---------→ .-HPETE | ↑ ↑ | | Θ Θ | | | | |

| NSAIDs .- Lio9)"en!se in2i(ito | ↓ ↓ PGF/, PGI/> PGE/> PGD/> TXA/ LTA*> LTB*> LTC*> LTC*> LTD*> LTE*

Met2o,;

1- "easure the weight of two rats to determine the dose of drugs.

2- 9n5ect one rat with acetylsalicylic acid in a dose of 188 mg0'g 9.4.

and another rat with normal saline 9.4.

- fter 1# min, in5ect both rats with 8.1 ml of fresh egg white into

the dorsal side of foot web of the rat.!- ;bserve for the absence of edema in the foot of the rat that

received acetyl salicylic acid, and the presence of edema in the rat

that did not receive the drug.

E9: Nee,s; E"" %2ite> !#et)' s!'i#)'i# !#i,

nee,'es> (!'!n#e:

1%


17/26

E9: No: 4

T2e E8!'0!tion of An!'"esi#s

nalgesics are classified as narcotics (acting on the /+ and causing

drowsiness li'e opiods* and non A narcotics (which act chiefly peripherally li'e acetyl salicylic acid*.

NSAIDs |

Θ ↓ Ne0oeti,e e'e!se → Inf'!!tion ----→ Me,i!tos e'e!se ↑ .-HT> PGs= Θ | | | OPIATES | | | Θ | ↓ ↓ E9#it!tion of t!nsission ne0ons ←------------- No9io0s

↓ Sti0'0s

P!in

Met2o,s;

1- hec' the weight of two rats, then observe and record the

following parameters:

- Ri"2tenin" ef'e9

- P!in ef'e9

- G!it

- Ee#tion of t!i' (or what is called st!0(s 2enoenon, whichoccurs due to stimulation of the spinal cord by the narcotics*.

2- 9n5ect one rat with morphine (9.4* in a dose of % mg0'g and the

other rat with acetyl salicylic acid (9.4* in a dose of 18 mg0'g.

- $echec' the above parameters in the two animals after 1# min. and

8 min. of in5ection, and record the difference in observation.

E9: Nee,s; C'!in" fo#es> N:S> o2ine> !#et)' s!'i#)'i#!#i,> S)in"es> nee,'es> (!'!n#e:

E9: No: +5

1@


18/26

Gene!' Anest2esi!

9tIs the absence of sensation associated with reversible loss ofconsciousness.

&uring induction of anesthesia, distinct stages occur with some agents that

include:

St!"e +;


19/26

Effe#ts of Anti#on80's!nts

Pin#i'e;

+trychnine is a /+ stimulant that interferes with the central inhibitory

processes with a powerful toxic effects.

Ai;

To show the convulsive rigidity in rats 0 mice when strychnine is

in5ected subcutaneously and to evaluate the action of anticonvulsants.

Met2o,s;

1- hec' the weight of two mice 0 rats.

2- 9n5ect one animal with normal saline 9.4 and mar' it as control, and

another animal with 4henobarbital 9.4 in a dose of #8 mg0'g.

- 1# minutes later, in5ect both animals with 2.# mg0'g of strychnine

+..!- $ecord the observations and time of occurrence in a table form.

E9: Nee,s; St)#2nine> P2eno(!(it!'> N:S:> Nee,'es> !n, B!'!n#e:

E9: No: +/

1D


20/26

Effe#ts of Antiei'eti#s

Pin#i'e;

7idocaine is a local anesthetic that is well 'nown to cause tonic Aclonic sei3ure.

Ai;

To determine the action of dia3epam as antiepileptic in the chemical

model of generali3ed convulsive St!t0s Ei'eti#0s in rats.

Met2o,;

1- hec' the weight of two rats 0 mice.

2- 9n5ect one animal with normal saline 9.4 and mar' it as control, and

another animal with dia3epam 9.4 in a dose of 2 mg 0 'g.

- 1# minutes later, in5ect both animals with 7idocaine 9.4 in a dose of

1#8 mg 0 'g.

!- ;bserve and record the following parameters:

!- T2es2o', of sei@0e:

(- D0!tion of sei@0e:#- T2e #!,io J esi!to) st!t0s:

,- De!t2 if o##0:

E9: Nee,s; Li,o#!ine> Di!@e!> N:S:> Nee,'es> !n, B!'!n#e:

E9: No: +$

28


21/26

DOSAGE FORM

I- INTERNAL PREPARATIONS

A- Ente!' C2e%!('e> Effe8es#ent:

• C!s0'es; O,in!)> S0st!ine, e'e!se

• Po%,e

/- Li0i, fo0'!tions

• A0eo0s; So'0tion> S)0

• A'#o2o'i#; Tin#t0e> E'i9i

• S0sension & "e'

B- P!ente!' !,inist!tion

+- In?e#tions; Ao0'es> Vi!'s

/- In2!'!tions; G!s> Aeosol

II- EXTERNAL PREPARATIONS

A- S3in Pe!!tions ; Ce!> Ointent> Lotion> P!int>

D0stin" o%,e

B- E)e> E!> Nose e!!tions ; Dos> Ointent

C- Re#t!' & V!"in!' e!!tions ;

Re#t!' s0osito)> 8!"in!' s0osito)


22/26

T!('etsTablets are solid dosage forms in which the drug is compressed with

pharmacologically inert substances called EG949E/T+. Tablets are

circular in shape with flat or convex surface, and film coated to improve

their appearance and stability and to mas' unpleasant taste.

A,8!nt!"es of t!('et fo;1. 4recision of dosage

2. &urability of physical characteristics

. +tability of chemical and physical activity of drugs

!. onvenience of administration.

C!s0'es apsule is a solid dosage form in which the drug is provided with

gelatin shell. =sed for drugs with unpleasant taste.

Mi#o-en#!s0'!tion


23/26

lear, pleasantly flavored liuid preparation of potent drug. The

vehicle contains high proportion of alcohol, sugar, or glycerol.

S0sensionBine solid particles of a drug suspended in aueous solution.

Ge' +emi-solid aueous preparation prepared with the aid of gelatin.

P!ente!' e!!tions

In?e#tions;+terile, pyrogen free solutions or suspensions used to produce rapid,

locali3ed, or prolonged action. They are of two types:

1. mpoule: provide a single dose

in5ection.

2. 6ial: provide multiple dose in5ections.

In2!'!tions;1. as: e.g. volatile anesthesia (nitrous oxide, ether*

2. erosol: particles dispersed in a gas under pressure to deliver the

drug directly into the respiratory tract.

E9ten!' e!!tions They are applied topically on the s'in and incorporated in a vehicle(base* that affects the degree of s'in hydration and influence the drug

penetration.

+'in preparations may ta'e one of the following forms:

Ce!

6iscous emulsion that is either aueous (oil-in-water* or oily (water-in-oil* and used as a vehicle for water-soluble or lipid-soluble drugs

respectively. reams are used to moisten the s'in and produce a cooling

effect as water evaporates.

Ointent They are semi-solid, greasy preparations for application to the s'in ormucous membranes. The base is usually anhydrous and immiscible with

s'in secretions. ;intments are insoluble in water and may be used as

emollients or to apply suspended or dissolved medicaments to the s'in.

2


24/26

Lotion ueous solution or suspension used to cool and relieve pruritis in

acutely inflamed lesions.

P!ints 7iuid preparation applied with a brush to the s'in or mucous

membranes.

D0stin" o%,e "ixture of two or more substances in fine powder form, used to reduce

friction and absorb moisture.

E)e> E!> !n, Nose e!!tions +terile isotonic solutions or suspensions, and may ta'e one of the

following preparations: Eardrops, Eye drops, Eye solutions or lotions, and Eye ointments.

Re#t!' & 8!"in!' e!!tions

Re#t!' s0osito) +olid dosage form shaped for insertion into the rectum. The

formulation is designed to melt at body temperature.

In,i#!tions fo e#t!' s0ositoies;1. =nconscious patient

2. +ever nausea

. =ncooperative child

!. 9rritant drug to stomach.

+olutions may also be given per rectum by the mean of enema.

V!"in!' s0osito)


25/26

E9: No: +*

PRESCRIPTION KRITING

I- Do#to ; /ame, Jualifications, /o. of registration.

II- P!tient ; /ame, ge, &iagnosis.

III- S0es#ition < R9 = ; $ecipe (treat with*

IV- Ins#ition ; &rugs

+- D0" n!e


26/26

VI- L!(e' ; &irections to the patient, e.g.

- to be ta'en at night (ypnotics*

- to be ta'en before meal (mpicillin*

- to be ta'en after meal (spirin*

- to be ta'en on need (;0/* (ngised*

VII- Si"n!t0e & D!te

Do#tos N!e

M:B:C2:B

No: of e"ist!tion

P!tients N!e;

A"e; D9;

Rx

+- D0" n!e> fo

manual of laboratory experiments-old

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