Proposal of Method Development for Stability- Indicating Assay test of Aspirin tablet (81mg)

!b"ective To create a stability-detecting assay for tablet Aspirin (81mg) by using HPLC.

Scope

sing systematic approac! to de"elop a ne# HPLC analytical met!od for creating a stability indicating assay test to separate Aspirin from its degradation product (salicylic acid) meeting t!e follo#ing parameters$ t!e column efficiency (%) is not less t!an &''' t!eoretical plates by optimi(ing pH) t!e tailing factor (T) is not more t!an 1.* by optimi(ing pH and mobile p!ase composition) t!e capacity factor+ ,-+ is not less t!an 1.' and not more t!an 1' by optimi(ing mobile p!ase composition) resolution factor (.) is not less t!an /.' by c!anging organic sol"ent and flo# rate. .01 s!ould not be more t!an /2.

Information of the sample

$1

Str%ct%re and Molec%le &eight of Aspirin

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C3H845

18'.16g7mol

&./.

Sol%bility 8 Aspirin is soluble in 9t!er+ met!anol+ c!loroform+ et!anol and slig!tly soluble in :ater.

&.&.

p'a ( at (/*;C) &.53

$)

Stability Degradation Prod%ct and Imp%rities T!e main degradation product is /-!ydro<yben(enecarbo<ylic acid (salicylic acid).

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*eference of analytical methods by +P,Compendium 1 . 0P Column C18 #it! L1 Pac,ing. >P C18 #it! L1 Pac,ing. = /8' nm >uffer7?obile P!ase /g of sodium 1- !eptanesulfonate in a mi<ture of 8*'ml of #ater and 1*'ml of acetonitrile+ and ad@ust #it! glacial acetic acid to pH &.5. Bn a flas, #it! a ground-glass stopper+ dissol"e 1.''' g in 1' mL of et!anol (36 per cent) .. Add *'.' mL of '.* ? sodium !ydro<ide. Close t!e flas, and allo# to stand for 1 !. sing './ mL of p!enolp!t!alein solution . as indicator+ titrate #it! '.* ? !ydroc!loric acid.

/ .

/&A nm

>ased on t!e pre"ious e<isting met!ods and t!e p!ysical- c!emical properties of t!e analyte+ .PHPLC is selected.

A ne& +P,- analytical method development

.1

+P,- Mode Selection

.e"erse P!ased HPLC met!od is selected for t!is stability because t!e analyte is relati"ely polar and soluble in 9t!anol+ ?et!anol and #ater.

.#

-ol%mn Selection

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According to t!e structure+ Aspirin is relati"ely polar compound and a small molecule. 0alicylic Acid is bot! t!e impurity and degradation product (0ee &.5 for 0alicylic acid structure and properties)) t!erefore+ t!e re"ersed p!ase column is c!osen. 0pecifically+ L1 pac,ed C18+ #it! *Cm in diameter+ t!e s!ort *'mm column+ sp!erical in s!ape+ a pore si(e to be at least 1*'A for compounds t!at !a"e a molecular #eig!t of D/''' is selected+ #!ic! is e<pected to pro"ide acceptable E-+ Tr+ resolution+ tailing factor and .01.

.$

Detector Selection

According to t!e structure properties+ #e select a F detector+ because Aspirin and its degradations !a"e lots of functional group (CG4 and ben(ene ring) #!ic! !a"e strong absorbance under F.

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.)

!ptimi/ation Mobile Phase

According to t!e structure and solubility properties+ t!e polar mobile p!ase is selected.

*.5.1.

4ptimi(ation Capacity Hactor ,-

An initial run mobile p!ase #it! *'$*'G met!anol$ #ater is c!osen+ t!e organic sol"ent concentration is decreased by 1'2 #!ic! is gi"en in t!e follo#ing table. -omposition 1. /. &. 5. *. Methanol *'2 5'2 &'2 /'2 1'2 0ater 1lacial Acetic Acid 532 *32 632 A32 832 12 12 12 12 12

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After testing t!ese fi"e mobile p!ase combinations (eac! one by / in@ections of eac! combination) t!e capacity factor map is generated+ and t!e optimi(ed combination can t!en be selected to gi"e E-G1. Higure 1 is a demo e<ample of capacity factor map.T!e optimi(ed mobile p!ase is furt!er test.

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*.5./. 4ptimi(ation of 0electi"ity Hactor I After capacity factor ,- is an optimi(ed+ a different organic sol"ent #it! t!e same sol"ent strengt! can be selected to optimi(e t!e resolution if necessary+ suc! as AC% 7 :AT9.+ and THH7 :AT9..

*.5.&. 4ptimi(ation of T!eoretical Plate % Bf necessary+ Plate number can be optimi(ed by c!anging pH+ suc! as using p!osp!ate buffer.

*.5.5 4ptimi(ation of Tailing Hactor+ T T!e tailing factor can be optimi(ed by c!anging pH using buffer or adding glacial acetic acid.

T!e column can be c!anged if t!e all optimi(ation failed.

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. ) Determination of 2 and 0or3ing -oncentration 0toc, preparation 0ee appendi<. Standard c%rve for p%re Aspirin 1ilute t!e standard solution to fi"e different concentrations. Transfer t!e standard solution to /*ml flas, and dilute #it! mobile p!ase. Pipetting 4ol%me (ml) -oncentration (ppm)

1 / & 5 *

/.* * 1' 1/.* 1*

*' 1'' /'' /*' &''

T!e = ma< is determined by scanning t!e /'' ppm solution using t!e F lig!t scanning mode. Bf t!e linear relations!ip bet#een t!e concentration and absorbency is not satisfied( at least t#o nines for sJuare .)+ t!e concentration range #ould be ad@usted base on t!e test data. Higure / is a demo e<ample of standard cur"e for #o,ing concentration determination.

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54 Meas%rements for standard -%rve 67ample

. . 8he 9inal !ptimi/ation After t!e indi"idual met!od optimi(ation (i.e. mobile p!ase+ #or,ing concentration+ HPLC settings)+ t!e met!od #ould be "alidated.

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. : Method Development *eport is prepared

Appendi<$ 1. 0ample preparation QC Mix$ :eig! 81 mg of APB+ e<cipients 1'' mg+ 0alicylic Acid /mg+ and transfer to /'' ml "olumetric flas,+ add *'mL mobile p!ase+ s!a,e it mec!anically for about * minutes+ and sonicate it for *minutes. 1ilute #it! mobile p!ase to "olume and mi<. Pass a portion of t!e solution t!roug! a filter !a"ing a -* micron or finer porosity+ discarding first * ml of t!e filtrate. se t!is clear filtrate as t!e assay preparation. Aspirin Standard Stock$ :eig! *'mg Aspirin .0+ and transfer to 1''ml "olumetric flas,. 1ilute #it! mobile p!ase to "olume and mi<. Hurt!er dilution is based t!e #or,ing concentration determined from t!e standard cur"e.

/. 0ample run procedure by HPLC /.1 0etup instrument+ /./ 0etup B?+?0+00?+ /.& After a single in@ection setup running time /.5 0tart at blan, (purified #ater 1''2) by 1 in@ection /.* .un KC mi< sample by / in@ection for indi"idual met!od optimi(ation. /.6 At t!e final met!od optimi(ation+ run KC mi< sample by * in@ection.

&.

Parameters (,+ %+ T+ .) of HPLC is calculated by 9mpo#er.

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