Pharmaceutical Analysis 2008PHM Laboratory Manual_2014 REPORT

8/9/2019 Pharmaceutical Analysis 2008PHM Laboratory Manual_2014 REPORT

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Pharmaceutical Analysis 2008PHM Laboratory Manual

General information and background:

Pharmaceutical analysis is a course intended to describe both theoretical andpractical aspects of method validation and spectroscopy. The three laboratory

experiments you will be undertaking during the course aim to give you a hands

on experience in such validations as well as measuring and interpreting

spectroscopic results.

Laboratory Safety Information: Please refer to provided notes in the Laboratory.

Method Validation

Method validation pertains to measuring and controlling the uality of analytical

methods and seeks to address such uestions as correct identity of drug in

formulation! whether the " stated composition of a drug present in a formulation

is correct and what concentrations of impurities are in drugs to name a few.

# $riteria are used to %udge the uality of an analysis method used to test

a drug and include

# Identifcation tests!

# Qualitative tests or imurities

# Quantitative tests o the active moiety in samles o a dru!"

substance or dru! roduct#

Please see the printouts in the laboratory for a description of the &nited 'tates

Pharmacopoeia (&'P) and *ritish Pharmacopoeia (*P) standards applying to

method validation applicable to the experiments here.

The laboratory aspect of +,-/-,,0P1M aims to introduce you to the physical

techniues of method validation and the types of euipment used in this process.

Structure of laboratory course$

'tudents will work in groups of 2 and perform + experiments! to be undertaken

in + single sessions.


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3verview of 4xperimental

'tations5 (6) %tation &

Concepts of measurement and reporting. This workstation introduces you some of

the physical techniues used in method validation/uality control relating to *ritish

Pharmacopoeia (*P) standards. Parameters such as disinte!ration, riability"

tablet hardness and dimensions will be determined on commercial ca7eine

tables.

(-) %tation2

Using analytical/spectroscopic method to analyse drug dissolution. This station will

examine ca7eine tablet properties in relation to dissolution. 8issolution

experiments will be performed under various conditions and temperatures using

di7erent p1 conditions and temperatures. 8issolution rates of ca7eine from tablets

will be determined by using &9:9;' spectroscopy.

(+) %tation'

Overview of analytical/spectroscopic measurements. 8etermination of compoundstability! concentration and reproducibility between drug batches is an important

consideration in *P standards. Typically a combination of spectroscopic methods

including (V)VI%" I*" +M*" Mass %ectroscoy ,M%-" Hi!h Perormance

Li.uid /hromato!rahy ,HPL/- and as /hromato!rahy ,/- are used in

tandem to determine the above.


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1orstation &

3isinte!ration" 4riability o atablet

Tablets consist mainly of >ller! except those in high:dose formulations. The most

common >ller is lactose! while the lubricant is typically magnesium stearate or

steric acid.


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De did not measure it due to the machine being out of service.

Part $ Cfter measurement of the three tablets place one of the tablets

in the tablet hardness tester (Aig -) for measurement of its hardness

=epeat this for the other - tablets


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Q# *ecord these values and reort the mean 6 %3 and 7*%3 orthese# ,2 Mars-

Values$

6) 6.E Mean F 6! 0 E

-) 60.6E ? '8 F 6! -6"

+) 6G.HE "='8F H! -,"

Q# 3id your results match the P standards9 :;lain# ,& Mar-

Cccording to *ritish Pharmacopoeia standards5

1C=8E4''F Hkg

Iroup results5

1C=8E4'' F 6!H6kg


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Plot this data using Microsoft 4xcel and paste this graph in the spaceprovided.

, G 6, 6G -, -G +, +G

6.

6.6

6.-

6.+

6.2

6.G

Mass (g)

5ime ,min-


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Q# 3id your results conorm to comendial standards9 :;lain# ,2Mars-

There is not a compedial test! meaning that there is no *P standard for them.

1owever! we think these tablets can be sold because the loss of weight was minimal.

Aigure 6. Tablet thickness test

Aigure - Tablet 1ardness tester.


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Aig +: Ariabilator

'# 5ablet 3isinte!ration#


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Aig +a. Tablet disintegration testing apparatus.


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1orstation 2

5ablet 3issolution

Please see appendix for the &'P EA standards on dissolution. The rate of dissolution

of a drug from a compounded product is an important consideration when designingtablets for various release rates. Many factors such as p1! drug stability to light!

heat! storage! level of impurities

in the sample all a7ect the rate of release of drugs from compounded tablets.

Please refer to the standard operating procedures for operation of the dissolutionapparatus.

Method5

(6) Place the tablet in a cell in the dissolution apparatus (Aig 2) (write down cellnumber).

Please note the clear plastic covers are to be inserted fat sie facing

above the tablet.

(-) ;nitiate stirring at +HN$ at 6,,prm (start your timer)

(+) Cfter exactly >ve minutes of stirring using the syringe provided withdraw

6.G mL of solution from the dissolution vessel. IMP>*5A+55 *eforesucking liuid into syringe please cover the tip of the syringe with the

>lters provided

(2) 'yringe out this volume into a &9 cuvette for measurement of &9:9;'absorbance.

=ecord your absorbance maximum (See supplied standard operating

procedure and ask your demonstrator if unsure). The &9:9;' spectrum of

ca7eine in water is shown in Aigure 2

(G) 'uck up the solution from the cuvette into the syringe and return this

to the bulk solution in the cell.

() =epeat the above measurement steps exactly every G minutes (up to+, minutes)


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&sing your absorbance maximum results complete the following table5

Time (min) Cbsorbance O -HG nm " 8issolution

G

6,-.+/ -.+++/ -.6/ -.H2G

Cverage F -.G2G0,.2"

6G

-,+.6-/ +.6-/ +.6-/ +.---

Cverage F +.6G6,,"

-G

+,+.6-/ +.6-/ +.6-/ +.---

Cverage F +.6G6,,"

+G

2,

2G

G,

GG

,

Q# Preare a dissolution rofle ,7 dissolution vs# time- !rah rom the dataobtained usin!

Microsot :;cel# Past your !rah in the sace rovided# ,? Mars)

Q# 1hat inormation can be obtained rom this !rah9 :;lain your ans


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Q# 1hat e@ect on


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Aig G (C) &9:9;' 'pectrum of $a7eine in water (*) $hemical structure of ca7eine.

Aigure ;=:spectrum of ca7eine


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13

1orstation '

>vervie< o analyticalBsectroscoic measurements inharmaceutical analysis#

;n this workstation you will use spectroscopic techniues such as &9:9;' to

gain an understanding of the methods used for measurement of accuracy! precision!

range! robustness and speci>city of the method.

Method5


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Q# 1hat is the 7 recovery o yoursamle9 ,&Mar-

Q# 3efne the linearity o the method ,& Mars-

;t is a form of internal or relative accuracyK for a speci>c focal

point! it re@ects how well a system responds to a seuence of

dilutions in the proper matrix. Dhen the experiment was done with

ca7eine! it was diluted and it was possible to see how well the

system was responding.

Q# 1hat is the accuracy o the method9 ,& Mar-

Cccuracy is a measure of the agreement of a particular

measurement with the true (or accepted) value of the parameter

under the conditions.

This analysis was accurate

Q# 1hat is the recision o the method9 ,& Mar-

The precision of a method is generally approached in terms of

either repeatability or reproducibility. Precision is the closeness of

agreement among test results obtained under prescribed conditions

The analysis showed itself imprecise

Q# 1hat is the Limit o 3etection9 ,& Mar)

The limit of detection! expressed as the concentration! or the

uantity! is derived from the smallest measure! that can be detected

with a reasonable certainty for a given analytical procedure.

Q# 1hat is the Limit o Quantifcation9 ,& mar-


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;t is the lowest concentration at which the analyte can not only

be reliably detected but at which some prede>ned goals for bias and

imprecision are met.

city and sensitivity as well as its

applicability in formulations with multiple CP;Qs or very low dose.


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5ablet3imensions

:;tra .uestions1orstation &

Tablet 6 Tablet - Tablet + Tablet 2 Tablet G

'ie 2.-6mm 2.-mm 2.-0mm 2.6Hmm 2.+-mm

Hardness


Aorce

Cpplied

6G.E 60.E 6H.HE 6.GE 6G.,E

4riability ) E.* G Tablets usedTime Tablets Deight

, Mins 6.G2

G Min 6.G6

6, Min 6.2

6G Min 6.2

-, Min 6.2,

-G Min 6.+

+, Min 6.+-

3isinte!ration


8isintegrationTime 6,min +Hsec 6-min Gsec 6+min G- sec 66min -Gsec 6-min 6,sec


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%ho


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3ur results5

A=;C*;L;T


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1orstation 2Q 1hat are some o the maFor sources o error durin! this e;eriment9:;lain your anslter

Time

*ecker (itJs not a precise instrument)

4uipament ( not calibrated properly)

1uman error

Q2# Gou are undertain! a dissolution study o a samle containin! thesodium salt o asirin# 1hat e@ect


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1orstation '

Q riey describe the rocedure o your method validation

usin! (V)visible sectrohotometer#,' Mars-

The method of validation includes verify the precision of the

method! the accuracy! and how is the graph and if it is showing the

result that was expected.

Q2# 3iscuss your results


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;t is used to determine functional groups in molecules. ;t is also

important to pharmaceutical uality control area because with ;=

spectroscopy it is possible to determine functional groups in

molecules.

Documents

Pharmaceutical Analysis 2008PHM Laboratory Manual_2014 REPORT