Glenn Kennedy's validation lecture

1
Decision Limit Detection Capability
2
CCa CCb
3
Non-Compliant lt----gt Compliant
4
Banned substances

• Zero tolerance
• Not detectable
• Detection Limits ??????????????????????????
• ???????????????????????????????????
• ??????????????????????????? Minimum
• Required Performance Limit MRPL

5
Banned substances

• ????????????????????????????????????????????????
  ??? MRPL ???
• ?????????????????? Non-compliant
  ???????????????????????????????? gt CCa

• ????????????????????? 99

6
New analytical limits
X
CCa - Decision Limit
Limit of detection
X
CCb - Detection Capability
Limit of quantification
7
CC? ??? CC?

• CC ????? critical concentration

• ? ????? alpha error
• ??????????????????????????????
  banned drug ?????????? ?????????????????????
  banned drug ??????????

False positive result
8
CC? ??? CC?

• ? ????? beta error
• ?????????????????????????????????
  banned drug ?????????? ?????????????????? banned
  drug ??????????

False negative result
9
CC? ??? CC?

• ???????????????????????????????????????????????
  ????????????????????????????????????????????????
  CC? ??? CC?

10
a- and ß-errors for banned compounds
a-error 1 b-error 5
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CCa
?????????????????????????????????????????
? ????????????????????????
12
Frequency
Signal
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Frequency
Signal
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????????? 2.33 SD
Frequency
a 1
CCa
Signal
15
blank samples ????? 1 ?????????????????? gt CCa
Frequency
CCa
Signal
16
???????? false positive results (gt Zero)
a-Error 1
Frequency
CCa
Signal
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Decision Limit - banned substances
CC? ???????????? 0 ??????????? MRPL
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Frequency
CCa
Signal
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?????????????????????????????? CCa
Frequency
CCa
Signal
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Frequency
CCa
Signal
21
Frequency
CCa
Signal
22
Banned substances
?????????????? CC? False negative rate 50
23
???
????????????????? banned drug ??????????????????
95 ????????????????????????????????
???
?????????????? Detection Capability
(CCb)
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CCb-banned substance
???????????????????????????????????
banned drug ??????????????? ??????????????? 95
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Frequency
CCa
Signal
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Frequency
CCa
Signal
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Frequency
CCa
Signal
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Frequency
CCa
CCb
Signal
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Banned substances
?????????????? CCb False positive rate
???????? False negative rate 5
(??? CCb ?????? lt MRPL)
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???????? CCa ??? CC?-banned drug

• ??????????? 2 ????
• 1. ????? fortified sample blank 3
  ??????????? ????
• 2. ????? sample blank

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???????? CCa - ??????? 1

• Fortify negative matrix ??????????????
• 1.0, 1.5 ??? 2 ??????? MRPL
• ????????????????????? 7 ???
• ??????????????? 2 ????? ??????????????????
• ???????????????? signal (??? Y) ???????
• ?????????? fortify (??? X)

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Calibration line
Signal
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?
????? SE ??? intercept
Calibration line
?
Signal
?
1.0 x MRPL
2.0 x MRPL
1.5 x MRPL
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??? SE ???? 2.33 (1, one sided)
?
Calibration line
?
Signal
?
1.0 x MRPL
2.0 x MRPL
1.5 x MRPL
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CCa ??????????????????????? signal
??????????????? (Intercept) (2.33 x SE of the
within laboratory reproducibility of the
intercept)
CCa ( 2.33SEintercept ) / slope
?
Calibration line
?
Signal
?
1.0 x MRPL
2.0 x MRPL
1.5 x MRPL
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???????? CCb ??????? 1

• Fortify negative matrix ????????????????????????
  ???????? MRPL ???????????????????? ???
• ???????????????? signal (??? Y)
  ????????????????? fortify (??? X)
• CC? ( 2.33SEintercept 1.64SECC? ) / slope

37
CCb ??????????????????????? Signal
??????????????? (signal??? CCa) (1.64 x SE ???
W/L reproducibility at CCa)
?
Calibration line
?
Signal
?
CCa
0.33 x MRPL
1.00 x MRPL
0.66 x MRPL
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???????? CCa - ??????? 2

• ????? sample blank ? 20 ???
• ????? S N ratio
• CCa ??????? 3 ??????? S N ratio

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???????? CCb ??????? 2

• ????? fortified sample blank ???????
• ?????????????? CCa ? 20 ???
• CCb CCa 1.64 S.D.

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???????? CC? ??? CCb ??????? 3

• ????? sample blank 20 ??? ?????????????????
  response ( ?n) ??? SD??? response (SDn)
• ????? fortified sample blank 21 ??? ????? RSD
  ??? response (RSDs)
• slope ??? calibration curve a

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???????? CC? ??? CCb ??????? 3

• CC? 2.33 (SDn) / a
• CC? 2.33 (SDn) 1.64 (?n) (RSDs)
• a 1 1.64 (RSDs)

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???????????????????????????????????? ????????
MRPL (???????????) ??????????

• Chloramphenicol (MRPL 0.3 µg/kg)
• ????? sample blank ??? fortified sample blank
  ??????????????????? ?????? 0.10,
  0.20, 0.30, 0.45, 0.60 µg/kg

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???????????????????????????????????? ????????
MRPL (???????????) ??????????

• ????????????????????? 7 ???
• ??????????????? 2 ????? ??????????????????
• ??????? accuracy ??? precision

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????????? 1
Q. What is CCß? A. lt 0.1 µg/kg
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????????? 1
Q. Is method fit-for-purpose? A. Yes. CCb is
less than MRPL.
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????????? 2
Q. What is CCß? A. 0.20 µg/kg
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????????? 2
Q. Is method fit-for-purpose? A. Yes. CCb is
less than MRPL.
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????????? 3
Q. What is CCß? A. 0.30 - 0.45 µg/kg
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????????? 3
Q. Is method fit-for-purpose? A. No. CCb is
greater than MRPL.
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Licensed substances

• ????????????????????????????????????
• ????????? ( Maximum Residue Limit MRL )
• ???????????????????????????????????????????
• ????????? MRL ???

• ?????????????????? Non-compliant
  ???????????????????????????????? gt CCa

• ????????????????????? 95

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a- and ß-errors for licensed compounds
a-error 5 b-error 5
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1.64SPL
1.64SS
CC?
PL
CCa
Frequency
a
?
XPL
XS
Signal
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???????? CCa ??? CC?-licensed drug

• ??????????? 2 ????
• 1. ????? fortified sample blank 3
  ??????????? ????
• 2. ????? fortified sample blank
  ?????????????????????

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???????? CCa - ??????? 1

• Fortify negative matrix ??????????????
• ???????????? PL ???????????????????? ???
• ????????????????????? 7 ???
• ??????????????? 2 ????? ??????????????????
• ???????????????? signal (??? Y) ???????
• ?????????? fortify (??? X)

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???????? CCa - ??????? 1
CC? ??????????????????????? Signal
??????????????? (signal ??? PL) (1.64 x SE ???
W/L reproducibility)
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???????? CCb ??????? 1

• Fortify negative matrix ??????????????
• ???????????? PL ???????????????????? ???
• ????????????????????? 7 ???
• ??????????????? 2 ????? ??????????????????
• ???????????????? signal (??? Y) ???????
• ?????????? fortify (??? X)

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???????? CCb ??????? 1
CCb ??????????????????????? Signal
??????????????? (signal??? CCa) (1.64 x SE ???
W/L reproducibility at CCa)
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???????? CCa - ??????? 2

• ????? fortified sample blank ? 20 ???
• CCa MRL 1.64SD

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???????? CCb ??????? 2

• ????? fortified sample blank ???????
• ?????????????? CCa ? 20 ???
• CCb CCa 1.64 S.D.