3 Implementation and validation of analysis methods
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1 3 Implementaton and valdaton of anal method 3. Preface When mplementng new method bacall three cae can be dfferentated: - Implementaton of offcal method (nternatonall approved, valdated method, e.g. method of the 64 LFGB / German Food, Feed and Commodte Act, the Analtca EBC or the MEBAK) - Modfcaton of alread etng method - Implementaton of new method that requre comprehenve valdaton. In trace anal the approach dffer from the other method lke content determnaton, phcal meaurement etc. In addton, content determnaton can be dfferentated n calbrateable method, at whch pecfc reference ubtance are avalable for addton tet and uch, for whch that not poble, e.g. foam meaurement or vcot determnaton. 3. Term, Defnton Work area The work area defned a concentraton area of an analte n an anal oluton, n whch the tet procedure provde relable reult n contant accurac. Calbraton functon Functonal correlaton between meaurable dmenon and concentraton to be determned. 30
2 The regreon anal provde a calbraton functon ( = a + b n cae of a lnear frt degree functon) wth the followng charactertc: - Slope b (meaure for the entvt of a method) - A ntercept a - Redual tandard devaton (dperon of the meaured data around the regreon lne) - Method dependng tandard devaton (abolute accurac meaure) - Method dependng coeffcent of varaton (relatve accurac meaure) Sentvt The entvt pecfe, to what etent the gnal of the tem to be meaured vare when changng one concentraton value. Recover rate The recover rate equal the quotent of recovered and added quantt multpled b 00 (pecfcaton n percent). If a recover rate of 00 % determned, the method free of devaton. Accurac Accurac a qualtatve term for the etent of appromaton of tet reult on a reference value, e.g. the true value. Accurac a uperordnate term for precon and truene. Precon Precon a qualtatve term for the etent of the mutual appromaton of tet reult ndependent from another n multple applcaton of a defned tet method under predefned condton. A dtncton drawn between repeatablt and reproducblt. Repeatablt Repeatablt the qualtatve term for the etent of mutual appromaton of tet reult under repettve condton. The et when under dentcal condton (ame peron n charge, ame equpment and ame laborator) wthn hort nterval a predefned anal method carred out everal tme ung dentcal ample. 303
3 Reproducblt The reproducblt a qualtatve term for the etent of mutual appromaton of tet reult under comparatve condton. The et, f meaurement reult are obtaned ung a defned anal method ung dentcal ample at varng nterval under dfferent condton (dfferent peron n charge, dfferent equpment, and varou laboratore) Truene Truene a qualtatve term for the etent of appromaton of the epected value of the tet reult on the reference value, wherea th can be the true or proper value. Detecton lmt The detecton lmt (DL) the lowet content of an analte n a ample that can be detected wth hgh predefned probablt. Lmt of quanttaton The lmt of quanttaton (LO) correpond wth the lowet analte content that can be quantfed wth a predefned precon. Specfct The analtcal pecfct the ablt of a tet method to olel detect the ought analte, wherea other element preent n the ample do not nterfere wth the tet reult. The MADEL goodne- of- ft- tet Calculator check of the calbraton data on lneart under conderaton of the redual tandard devaton a well a the varance of the lnear calbraton functon and the econd-degree calbraton functon. Outler tet b redual anal Calculator check of the calbraton data on outler under conderaton of the redual tandard devaton. 304
4 3.3 Implementaton of new method: approach ew method Modfcaton of etng method offcal method Content determnaton Trace anal Content determnaton Trace anal Content determnaton Trace anal ca nca ca nca Specfct Lneart Truene Precon Recover Lmt of quanttaton Detecton lmt Robutne ca = calbrateable method nca = non calbrateable method 3.4 Montorng/ualt aurance For montorng the qualt of anal method varou ntrument can be ued. Interlaborator tet/comparon anal On one hand the hall how tematc devaton; on the other hand the hall conve ecurt for nvolved emploee. The reult of the nterlaborator tet are beng evaluated and repectve meaure are beng deduced. 305
5 Control chart The prncple of control chart an optcal llutraton of meaured data takng a ba - ualt objectve (pecfed value of the reult of control ample) - ualt bound The latter are dfferentated between warnng lmt, whoe ndvdual eceedance beng tolerated, and control or acton lmt, whoe eceedance caue meaure. Control chart are manl ued there, where the anal precon and the truene of a reult of ver hgh mportance, e.g. crtera of ale anal, that form the ba for bllng. Control ample Varou categore of control ample can be ued: - Blank tet that defntel do not contan the ought analte - Reference materal wth defned concentraton of ndvdual analte, e.g. CRM-ample (certfed reference materal) - Reference materal wth defned properte, e.g. calbraton ol for calbraton of the vcometer or tandard malt for calbratng the malt Frablmeter. 306
6 Mathematcal Bac 3.5. Regreon anal Aular quantte ) ( = ) ( = = ) ( = 3 3 = 4 ) ( 4 = ) ( ) ( = = Frt degree regreon Slope b = A ntercept b a = Correlaton coeffcent ( )( ) ( ) ( ) = ) ( r Redual tandard devaton = Method tandard devaton b o =
7 Method coeffcent of o varaton Vo = 00% Lower lmt of work area p = o t ( p ) + + b Second degree regreon A ntercept a = b c Coeffcent b = c 3 Coeffcent Redual tandard devaton c = ( ) ( a b c ) = Sentvt E = b + c Method tandard devaton o = E 308
8 3.5. Lneart tet: MADEL adaptaton tet Redual tandard devaton for frt degree calbraton functon Redual tandard devaton for econd degree calbraton functon DS = ( ) ( 3) Degree of freedom f = Tet value DS PW = Comparon wth table value F (f =, f = 3, P = 99 %) If PW F, then no gnfcantl better adaptaton obtaned b the econd degree calbraton functon. The calbraton functon lnear Outler tet va F-tet of redual varance A a bac prncple, calbraton data mut be outler-free. The redual anal can be ued for detecton of outler. For th purpoe at frt the calbraton lne calculated va redual tandard devaton A baed on all par of varate. After elmnaton of a upect outler par of the data a new calbraton lne calculated wth the redual varance. The verfcaton conducted wth the F-tet. A 309
9 A Redual tandard devaton for calbraton functon ung all value A Redual tandard devaton for calbraton functon wthout upect outler value Tet value ( PW = A A ) ( ) A Comparon wth table value F (f =, f = A -, P = 95 %) If PW < F, then no outler et. A A 3.6 Lterature. W. Funk, V. Dammann und G. Donnewert, ualtätcherung n der Analtchen Cheme, VCH Verlaggeellchaft mbh, Wenhem, German 99 30
10 t-table f t (P = 95 %) t (P = 99 %) f = degree of freedom F-table for f = f F (P = 95 %) F (P = 99 %) f, f = degree of freedom 3
11 FOR Method valdaton Tpe of ample: Valdaton on: Method: Stattc charactertc Calbraton functon lnear econd degree polnomal other Precon (.3) r: R: Recover Addtve:.. % Addtve:.. % Lmt of quanttaton Meaurng range... Approved: Date/Sgnature.... 3
12 . Specfct Ye o Date/Sgnature.. Anal of ample olvent.. Anal of contamnaton/degradaton product.3. Comparon of peak form, retenton tme and UV-pectrum of tandard and ample.4. Precon Ye o Date/Sgnature.. Precon of devce: 6 tme determnaton of tandard.. Precon of devce: 6 tme determnaton of ample.3. Precon of method: 6 tme recondtonng of homogenou pattern.4. 33
13 3. Truene/Recover 3.. Recover, addtve equvalent to L determnaton tme. 3.. Recover, addtve equvalent to 0 L determnaton tme 3.3. Recover, addtve equvalent to. determnaton tme 3.4. Anal of certfed reference materal, CRM. determnaton tme 3.5 Comparon of varou etracton technque 3.6. Ye o Date/Sgnature 4. Lneart 4.. Lneart tandard: 5 concentraton between... and Lneart ample preparaton: 5 concentraton between... and Ye o Date/Sgnature 34
14 5. Meaurng range Ye o Date/Sgnature Detecton lmt Ye o Date/Sgnature 6.. Detecton lmt (Sgnal/noe 3:): Lmt of quanttaton Ye o Date/Sgnature 7.. Lmt of quanttaton (Sgnal/noe 0:):. 8. Robutne Ye o Date/Sgnature
15 9. Stem qualfng eamnaton Ye o Date/Sgnature 9.. Retenton tme of 9... Injecton precon of 9.3. Anal of blank tet
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