(ACCURACY) (1) (, error bias) (mean) xmean, , (true) (correct) . xi . Xmean Xi (true value).

(ACCURACY) (2) :) (random errors)) (bias) ( ). (accurate) (precise). (xmean ) ( , bias) (truness) . = (xi ) = (xi xmean) + (xmean ) = ( ) + (bias) = ( ) + ( )

(ACCURACY) (3) (TOTAL ERROR)Xmean = (bias)

S = ,

: (reference material) (, spiked) ,

(accuracy) (precision)

(ASSAY) ( , reference material)(t-test) () , / ( ) (t-test). , .

(ASSAY) () (1) . . , xi i. (intercept, a) , (b-1)x100 Sy/x .

(ASSAY) () (2)2. :%R=[Cspiked / (Co + C)] x 1003. , () .

(ASSAY) (1) , (spiked) . , .

(ASSAY) (2) (response factor) . (placebos), .

(1)- 9 , (. 3 x 3 ). 5 , 50 150% . 6 .

(2) : (%Recovery) . (accepted true value) ( ) .

- :- () (theoretical value)- (x) (assay value)- (percent recovery) (x/).100- , , , - .

, 98%-102% . 90%-110%. 4 . .

Accuracy Data Set (1 of 3)*

Amount Added (mg)AmountFound (mg)Percent Recovery0.00.0---50.250.4100.579.680.1100.699.9100.7100.8120.2119.899.7150.4149.799.5

(PRECISION) (1) - () () . . (dispersion) .

(PRECISION) (2) . , (normal distribution Gaussian)

(PRECISION) (3) () :- ( ): (mean) xmean- ( ): s SD s (range) .

(PRECISION) (4) : (Repeatability) (Reproducibility)

REPEATABILITY REPRODUCIBILITY (1): , , . , , , .

REPEATABILITY REPRODUCIBILITY (2): , , . , , , .- ( ) [ Intermediate Precision ]- ( ).

REPEATABILITY REPRODUCIBILITY (3) Within run precision: : ( ). (System Precision)

REPEATABILITY REPRODUCIBILITY (4) Between run precision: : . , . , :Within a day precision: Between days precision: ()

REPEATABILITY REPRODUCIBILITY (4) (Repeatability Limit r) :r = t x 2 x r

t Students = 95%, 1,96 r . :r =2,77 x r

REPEATABILITY REPRODUCIBILITY (4) (Reproducibility Limit R) :R = t x 2 x R

t Students = 95%, 1,96 R . :R =2,77 x R

- 9 (. 3 x 3 6 100% .- .- , , ,

(INTERMEDIATE PRECISION)

- . - :- - - - / (. )- .

- . - (standardization) , .. , AOAC, .

(2)

- ( ).- robustness (), , .

(1)

(s SD), (sr RSD) . . (uncertainty) .

(2)

: . 20 . . 6-7 ().

(3)

( ): 1) 2%2) .3) / .

(4)

- CHPB RSD :- 1,5% - 2,5% - - , 6 . . .

(5)

- RSD ( , CHPB):- 1% - 2% - 5% - 10%

(6)

:- , 10- .- , , ( ) / .

Repeatability & Intermediate Precision*GrandMean = 100.0RSD = 0.59%Mean = 100.5RSD = 0.24% Mean = 99.5RSD = 0.36%

Day 1Day 2100.699.5100.899.9100.198.9100.399.2100.599.7100.499.6

Intermediate precision - Japanese styleExpt No12345678Operator11112222GC12211221Column12121212

Experiments are conducted on different days with each analyst using each instrument/column combination in turnReport results, overall mean, overall standard and relative standard deviation, and 95% CI of sd

ReproducibilityDefinition: Ability reproduce data within the predefined precisionDetermination: SD, RSD and confidence interval Repeatability test at two different labs.

*

Lab 1Lab 2Lab 3Day 1Day 2Day 1Day 2Day 1Day 2Man 1Man 2Man 1Man 2Man 1Man 23 Prep3 Prep3 Prep3 Prep3 Prep3 Prep

HORWITZ %RSDR = CVR = 2(1-0,5 log C), C 10, . 1 mg/g 10-3

Horwitz Ratio (HorRat) (1)Horwitz Ratio (HorRat): %RSDR () Horwitz (%RSDR)HoRat 0,5: (, ),HoRat 2 : , )

Horwitz%RSDR = 2(1-0,5logC), C = mass fraction%RSDR = 2C-0.15SDR = 0,02C0,8495 %RSDR=(SD/C)x100 , C (Analyst 125, 385, 2000)SDR=0,22C C < 1,2x10-7SDR = 0,02C0,8495 1,2x10-7 C0,138SDR = 0,01C0,5 C>0,138

%RSDH

Horwitz Ratio (HorRat) (2) %RSDR %RSDr ( ) %RSDr = 0,66 %RSD%RSDR = 1,52 %RSDr HorRat AOAC, ,.

Typical expected results for Analyte concentration vs. Precision(The AOAC manual for the Peer-Verified Methods program)

Analyte Conc (%)Analyte RatioUnitRSD%1001100%1.31010-110%2.8110-21 %2.70.110-30.1%3.70.0110-4100 ppm5.30.00110-510 ppm7.30.000110-61 ppm110.0000110-7100 ppb150.00000110-810 ppb210.000000110-91 ppb30

(DETECTABILITY) (1)

/ . :) (Detection Limit, DL LOD)) (Quantitation Limit, QL LOQ) .

(DETECTABILITY) (2)

/ :1) ()2) .3) ( ).

(MEASUREMENT DECISION)

(DETECTABILITY) (3)

( ) :- : .- : .

(DETECTABILITY) (4)

: (x), (1-) . () .- () .

(DETECTABILITY) (5)

x: x = xo + k xo = (blank) ( / ) = k= 1- ( ) C = [ xo + k] / S S ( ) .

= , 1 = , = , = , =

(DETECTABILITY) (6)

0,5000,15870,02280,00620,00260,00130,0005k01,02,02,52,83,03,3

(DETECTABILITY) (7)

k = 3,3 = 0,0005 0,05% : : 3,3 . : 10 .

= 3. 50%.

, k = 6

(1)

, .- (visual evaluation): . , .

(1)

(signal to noise ratio, S/N): (baseline noise). () . S/N = 3,3 3 () LOD S/N= 10 LOQ.

(2)

(SD) (response) (slope) (b) : LOD = [3,3 x SD] / bLOQ = [10 X SD]/b ( ) SD :1) (blank)2) ( ).

(3)

3) , SD ( ) (SD ) .4) (Sa) ( ), , .5) (Sy/x) ( ), , .

HPLC LOD/LOQ

(4)

, .

, .

*

(Ruggedness) (1)( ROBUSTNESS) . (robustness) .

(Ruggedness) (2)( ROBUSTNESS) . .

(Ruggedness) (3) :- - -

(Ruggedness) (4) HPLC:- pH - - ( / )- -

(Ruggedness) (5) GC:

- ( / )- -

Robustness IIIInfluence of pH of elution on separation of amino acids by RP-HPLC

From: Waters, in: Stavros Kromidas, Validierung in der Analytik, Wiley-VCH

RobustnessElectropherograms under identical conditions by different analytical equipment

From: Dr. Michael Krmer, NOVARTIS, Basel, in: Stavros Kromidas, Validierung in der Analytik, Wiley-VCH

() (RUGGEDNESS)

(SENSITIVITY) (1) , . (slope) . , (dynamic range).

(SENSITIVITY) (2) (linear dynamic range). ( ) .

USP 23 System Suitability Requirements*

Parameters RecommendationsK In general k 2.0R R > 2, between the peak of interest and the closest potential interferent (degradant, internal STD, impurity, excipient, etc..)T T 2N In general N > 2000Repeatability RSD 2.0% (n 5)

(1)

(2)

HPLC

*