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Quantitative Validation2010 QP PDF
Quantitative Validation2010 QP PDF
REFERENCE STANDARDS:
Standard Information
Primary Standard
Primary Standard
Materials which are accepted without reference to other standards.
If the materials have undergone complete analytical characterization, their identity
must be proven (elucidation of chemical structure) and their purity must be sufficiently
high and stated (>99.0%).
The characterization of primary standards generally involves the following:
Elucidation of chemical structure by IR, UV, H-NMR, C-NMR, MS, CD etc.
Purity determination by HPLC,TLC,GC,GPC,DSC,residue of ignition, water content etc.
Assay: Titration,DSC,Chromatography.
It is acceptable that the manufacturing process of primary references standards
differs from the final processing of the drug substance.
Pharmacopeial Standards
Commonly used for certain tests and assayed to achieve accuracy and precision of analytical results required in
compendia monographs. Pharmacopeial standards are basically regarded as primary standards.
Working Standard
Working Standards
Materials are designed for daily use in instrumental analysis such
as routine quality control.
They are characterized by comparison with Primary or
Pharmacopoeia standards. Their purity corresponds to a "typical
batch".
Integration
End
Abs x dt
Start
Detector
Signal
Time
Measurement of Area:
Peak Integration
Data Bunching
Peak Start
Peak Apex
Peak End
Peak Height
Constructed
Baseline
Start of
chromatogram
Peak End
Peak Start
Retention Time
Measurement of Area:
Peak Integration: Data Bunching
15 = Minimum Number of points to define a peak
B = (WxS)/15
Bunching
B = BUNCHING FACTOR
W=PEAK WIDTH
S = SAMPLING RATE
Data
points
Measurement of Area:
Peak Integration - Peak Start
Slope 2 =
(B3-B2)/(t3-t2)
Slope 1 =
(B2-B1)/(t2-t1)
B3
Average
Slope
B2
B1
Threshold value
Measurement of Area:
Peak Integration - Peak End
Slope 2 =
(B3-B2)/(t3-t2)
Average
Slope
Slope 1 =
(B2-B1)/(t2-t1)
B3
B2
Threshold value
B1
(Touchdown)
Measurement of Area:
Peak Integration - Peak Apex
Slope 1 =
(B2-B1)/(t2-t1)
Slope 2 =
(B3-B2)/(t3-t2)
10
98.972
98.972
0 .0 4
100.00
0.04
8 0 .0 0
80.00
4 0 .0 0
60.00
0.02
AU
AU
0 .0 1
% Compos ition
6 0 .0 0
0 .0 2
0.01
40.00
% Compos ition
0.03
0 .0 3
0.00
0 .0 0
- 0 .0 1
-0.01
2 0 .0 0
20.00
-0.02
- 0 .0 2
0.00
0 .0 0
-0.03
- 0 .0 3
9 8 .0 0
9 9 .0 0
1 0 0 .0 0
1 0 1 .0 0
98.00
1 0 2 .0 0
99.00
100.00
101.00
102.00
Minutes
Min u te s
Peak Threshold
Peak Threshold=300
Peak Threshold=75
100.00
100.00
0.015
0.015
80.00
80.00
0.010
0.000
40.00
0.005
60.00
AU
% Composition
AU
60.00
0.000
40.00
-0.005
-0.005
20.00
20.00
-0.010
-0.010
-0.015
-0.015
0.00
70.00
71.00
72.00
73.00
0.00
70.00
74.00
71.00
72.00
Minutes
Minutes
11
73.00
74.00
% Composition
0.005
71.153
71.152
0.010
47.893
0 .0 3
45.253
43.741
0 .0 2
AU
0 .0 1
0 .0 0
- 0 .0 1
- 0 .0 2
4 2 .0 0
4 4 .0 0
4 6 .0 0
4 8 .0 0
5 0 .0 0
M in u t e s
0 .0 5
47.893
1 0 0 .0 0
0 .0 4
8 0 .0 0
45.253
0 .0 2
0 .0 3
6 0 .0 0
0 .0 2
AU
AU
0 .0 1
0 .0 1
4 0 .0 0
0 .0 0
0 .0 0
- 0 .0 1
- 0 .0 1
2 0 .0 0
- 0 .0 2
- 0 .0 2
- 0 .0 3
4 2 .0 0
4 4 .0 0
4 6 .0 0
M in u t e s
4 8 .0 0
0 .0 0
4 2 .0 0
5 0 .0 0
4 4 .0 0
4 6 .0 0
4 8 .0 0
5 0. 0 0
M in u t e s
1 00.0 0
57.786
0.0 05
52.316
52.316
0 .00 5
57.786
80.00
8 0.00
4 0.00
60.00
AU
% Composition
6 0.00
-0 .01 0
-0.0 10
40.00
-0.0 15
-0 .01 5
-0 .02 0
-0 .02 5
50 .00
52.00
54.00
Minu tes
5 6.00
2 0.00
-0.0 20
0 .00
-0.0 25
20.00
0.00
50.00
5 8.0 0
12
52.00
54.00
Minutes
56.00
58.00
% Composition
-0.0 05
-0 .00 5
AU
100.00
0.0 00
0 .00 0
% Composition
47.893
45.253
43.741
0 .0 3
43.741
Tangential Skim
Exponential Skim
0.04
0.04
0.03
0.02
0.02
AU
AU
0.03
31.163
31.453
31.453
0.05
0.01
0.01
0.00
0.00
-0.01
-0.01
-0.02
-0.02
30.50
31.00
31.50
32.00
32.50
30.50
33.00
31.00
31.50
32.00
32.50
33.00
Minutes
Minutes
Valey to Valey
Original
Valey to Valey
0.18
0.18
0.12
80.00
0.10
0.10
60.00
0.06
% Composition
0.08
62.087
0.08
AU
62.087
% Composition
60.00
0.06
40.00
0.04
66.548
0.04
66.548
40.00
61.419
AU
67.321
80.00
65.016
0.14
67.321
65.016
62.773
0.12
62.773
0.16
0.14
0.02
100.00
100.00
0.16
0.02
20.00
20.00
0.00
0.00
-0.02
-0.02
0.00
0.00
62.00
62.00
64.00
66.00
64.00
66.00
68.00
Minutes
Minutes
13
68.00
Asymmetric
5 % of Peak Height
Slope 2 =
Slope 2 =
(B3-B2)/(t3-t2)
(B3-B2)/(t3-t2)
Slope 1 =
Slope 1 =
(B2-B1)/(t2-t1)
(B2-B1)/(t2-t1)
B3
Average
Average
B3
Slope
Slope
B2
B2
B1
Threshold value
Threshold value
14
B1
PW=15
Area=23011
-10%
AUFS = 0.003
Peak width changed
Threshold set at 30
3.80
4.20
Minutes
4.60
3.80
4.20
Minutes
4.60
Thresh.= 20
Area=25660
Thresh.=300
Area=23087
AUFS = 0.003
-10%
Peak width set at 30 sec
Threshold changed
3.80
4.20
Minutes
4.60
3.80
4.20
Minutes
4.60
15
3.
2.
4.
5.
1.
16
6.
Login.
Instrument Method.
Sequence/Run Samples.
Process Data.
Review/Preview.
Result/Sign-Off.
FDA Guidance
Table 2. Holistic Validation of Computerized LC Systems
_________________________________________________________________
Initial Calibration--Linearity
Use at least 4 standard solutions.
Concentration range of standards must span anticipated results, plus safety margin.
Run standards daily before starting sample analysis.
Initial Calibration--System Precision
Calculate precision daily from at least 6 replicate injections of a standard solution
before starting sample analysis.
Running Calibration
Run a standard solution at specified time intervals or after a specified number of
sample solutions.
Data Processor (Integrator, Computer(
17
INJ VOL
No of Inj
Function
Method
Blank
20.0
Inject Samples
System
Suitability
20.0
Inject Samples
Std1
20.0
Clear
Calibration
5 Inject Standards
3
4
Std2
20.0
1 Inject Standards
Dilution
10.00
1.00000
1.00000
10.00
1.00000
1.00000
10.00
1.00000
1.00000
10.00
1.00000
1.00000
Report
LC Calibration Report
Report
Standard Comparison
Std1
20.0
Clear
Calibration
1 Inject Standards
10.00
1.00000
1.00000
Unk.1
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.2
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.3
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.4
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.5
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.6
20.0
Inject Samples
10.00
1.00000
1.00000
Std1
20.0
1 Inject Standards
10.00
1.00000
1.00000
1
2
Clear
Calibration
Calibrate
Blank
mAU
30.00
20.00
? ?
10.00
0.00
50.00
40.00
Sample
mAU
30.00
20.00
10.00
0.00
0.00
10.00
20.00
30.00
Minutes
18
40.00
50.00
Injection
System Peaks
Legitmate System Peaks
Originate from the Mobile Phase Components Going
Through Re-Equilibration
19
k' =
tR - t 0
k =
t0
Cs
Example: k(2
k(2) = 2
Step 1:
Step 1:
Equilibrium:
Cs=1
Cs=
1; Cm = 1
Equilibrium:
Cs=2
Cs=
2; Cm = 1
Step 2:
Injection of Vacancy:
Cs=1
Cs=
1; Cm=0
Cm=0
Cm
Step 2:
Injection of Vacancy:
Cs=2
Cs=
2; Cm=0
Cm=0
Step 3:
Re
Re--equilibration:
Cs=0
Cs=
0.5; Cm=
Cm=0
0 .5
CHROMATOGRAM
Step 3:
Re
Re--equilibration:
Cs=1
Cs=
1.33
33;; Cm=
Cm=0
0.67
t0
k=1
20
k=2
An Example for Real System Peaks in Gradients with Trifluoroacetic Acid (TFA) in the Mobile
Phase
Peptide Peak
Sample=Peptide
Dissolved in Water
TFA
Zoomed
ACN
Blank =Water
0.005
0.004
AU
0.003
0.000
6.477
4.274
2.373
2.731
0.001
18.274
0.002
-0.001
-0.002
-0.003
2.00
4.00
6.00
8.00
10.00
12.00
14.00
Minutes
21
16.00
18.00
20.00
22.00
24.00
26.00
28.00
30.00
250.00
2.748
nm
300.00
250.00
2.867
nm
300.00
250.00
3.180
nm
300.00
250.00
4.226
nm
300.00
300.00
Aromatic (not in
mobile phase)
Negative UV
spectrum
Cpd 2 - 2.188
0.000
1.00
1.20
1.40
1.60
1.80
2.00
2.20
2.40
2.60
2.80
3.00
3.20
4.226
0.005
Wavelength: 254.0 nm
2.748
2.867
AU
0.010
3.180
0.015
3.40
3.60
3.80
4.00
4.20
4.40
4.60
Minutes
Yes?
No
Mobile phase:
Multicomponent?
Adsorbed components (ion pair)?
Response in detector (background)?
Yes?
22
4.80
5.00
5.20
5.40
5.60
5.80
6.00
23
No of Inj
Function
Method
Dilution
Blank
20.0
1 Inject Samples
10.00
1.00000
1.00000
2 System
Suitability
20.0
1 Inject Samples
10.00
1.00000
1.00000
10.00
1.00000
1.00000
10.00
1.00000
1.00000
Std1
20.0
Clear
Calibration
5 Inject Standards
Std2
20.0
1 Inject Standards
Report
LC Calibration Report
Report
Standard Comparison
Std1
20.0
Clear
Calibration
1 Inject Standards
10.00
1.00000
1.00000
Unk.1
20.0
2 Inject Samples
10.00
1.00000
1.00000
Unk.2
20.0
2 Inject Samples
10.00
1.00000
1.00000
Unk.3
20.0
2 Inject Samples
10.00
1.00000
1.00000
Unk.4
20.0
2 Inject Samples
10.00
1.00000
1.00000
Unk.5
20.0
2 Inject Samples
10.00
1.00000
1.00000
Unk.6
20.0
2 Inject Samples
10.00
1.00000
1.00000
Std1
20.0
1 Inject Standards
10.00
1.00000
1.00000
Clear
Calibration
Calibrate
24
System Suitability
Sample Name:
Sample Type:
Vial:
Injection #:
Injection Volume:
Run Time:
AntiOX 10 2d
Unknown
2:A,2
1
2.00 ul
2.0 Minutes
Acquired By:
Sample Set Name:
Acq. Method Set:
Processing Method:
Channel Name:
Proc. Chnl. Descr.:
Date Acquired:
Date Processed:
System
Antiox_sampling_rate rsd 2D
Restech_Std_UPLC_10_2d_01
Antioxidants_UPLC 220nm
PDA Ch1 220nm@4.8nm
PDA Ch1 220nm@4.8nm
0.02
0.04
AU
0.06
Peak Results
Name
VIT E - 0.731
BHT - 0.193
0.08
ENICAMIDE - 0.303
0.10
Auto-Scaled Chromatogram
RT
EP USP
USP USP
Area Height Plate Plate
Alfa
Res Tailing
Count Count
1 BHT
0.193
97392 106748
1028
948
2 ENICAMIDE
0.303
88614
74300
1379
1419
3.84
1.34 3.391
4 IRGANOX1010 0.552
77254
44092
2229
2287
6.37
1.21 2.603
5 VIT E
59739
37644
4872
4754
4.03
1.18 1.441
64671
5171
5098
4.87
1.14 1.402
7 IRGAFOS 168
18010
6640
6561
5.64
1.09 1.407
3 IRGANOX3114 0.459
0.731
1.299
43447
0.00
0.00
0.20
0.40
0.60
0.80
Minutes
1.00
1.20
1.46
1.40
Project Name:
Test_Sampling_Rate
Date Printed:
16/04/2010
20:48:52 Asia/Tel_Aviv
PEAK BROADENING
25
System Suitability
is
Based on the Theory of
Chromatography
Mobile phase
Stationary
Phase
Abs
A
Area A = 300,000
Area B = 100,000
B - 94.776
K = C s/C m
AU
0.34
0.32
0.30
0.28
0.26
0.24
0.22
0.20
0.18
0.16
0.14
0.12
0.10
0.08
0.06
0.04
0.02
0.00
-0.02
A - 93.448
Chromatogram
Distribution:
90.50 91.00 91.50 92.00 92.50 93.00 93.50 94.00 94.50 95.00 95.50 96.00
Minutes
Time
26
Mixed--Mode Retention:
Mixed
Ion exchange Interaction
with Charged Sites
Mobile
Phase pH < 3
Si - OH
O- Si
O- Si
OH
O- Si
O- Si
OH
O- Si
OH
O- Si
O- Si
O- Si
OH
O- Si
O- Si
+
HN (CH 3 )
Base
O- Si
O- Si
OO- Si
O- Si
+
O - (CH ) HN
3 2
O- Si
OO- Si
O- Si
O- Si
OO-- Si
O- Si
Mobile
Phase pH > 3
Si O
Base
27
System Suitability
System Suitability
The checking of a system, before or during analysis of unknowns, to insure system
performance.
No sample analysis is acceptable unless the requirements for system suitability have
been met. (USP Chapter 621)
Plate Count, Tailing, Resolution
Determination of reproducibility (%RSD)
For %RSD < 2.0%, Five replicates
For %RSD > 2.0%, Six replicates
System Suitability "Sample"
A mixture of main components and expected by-products utilized to determine system
suitability
Whenever There is a Significant change in Equipment or Reagents, System Suitability
Testing Should be Performed (USP Chapter 621)
t1
t3
t0
TIME
.5
5
k' t1 = 1-0.5= 1
0.5
k' t1 = t1 - t0
t0
K' t2 = 2-0.5= 3
0.5
k' t3 = 5-0.5= 9
0.5
28
29
Application
BY ONE PEAK
PERFORMANCE CRITERIA
k' =
tR - t 0
k =
t0
Af =
B(10% h)
(10%
h)
TAILING FACTOR
0.020
Area:
USP Resolution:
USP Tailing:
K Prime:
Retention Time:
height
0.015
AU
Cs
Cm
28501
4.48
1.11
4.02
0.74
Tf =
(5% h)
A+B
2A
Width (50%)
0.010
0.005
10%h
A
0.000
Width(base)
0.710
0.720
0.730
0.740
0.750
0.760
0.770
Minutes
30
N = 16 (
tR
w
k' ( 2 )
k' (1)
tR(1)
tR(2)
EXPERIMENTAL RESOLUTION
Rs =
t R (2)
1/2
-t
(w
R(1
+w
)
2
t0
w1
w2
Rs =
t R (2)
1/2
(w
-t
1
R(1 )
+w
2)
tR(2)
tR(1)
t0
w1
31
w2
ADJUST N (Plates)
100:1
10:1
Dolan
32
UPLC
HPLC
SELECTIVITY vs EFFICIENCY
Rs =
t R (2) - t R(1)
Same in both
1/2 (w 1 + w 2) cases
LOW SELECTIVITY (
)
HIGH EFFICIENCY (N)
HIGH SELECTIVITY (
)
LOW EFFICIENCY (N)
33
SELECTIVITY vs EFFICIENCY
Rs =
t R (2) - t R(1)
same in both cases
1/2 (w 1 + w 2)
HIGH SELECTIVITY (
)
LOW EFFICIENCY (N)
LOW SELECTIVITY (
)
HIGH EFFICIENCY (N)
0.50
0.40
0.30
AU
2.5 m
2.1 x 20 mm
HPLC
0.20
0.10
0.00
0.20
0.80
0.60
AU 0.40
0.40
0.60
Minutes
0.80
1.00
1.20
1.40
1.20
1.40
5 m
2.1 x 50 mm
0.20
0.00
0.70
0.50
1.80
1.60
2.00
0.40
1.548
0.10
1.231
0.20
1.091
1.146
0.30
1.002
AU
0.60
1.7 m
2.1 x 100 mm
UPLC
1.00
Minutes
4 - 1.652
0.90
0.80
0.80
2 - 0.803
0.60
3 - 0.901
0.40
1 - 0.707
0.20
0.00
0.20
0.40
0.60
0.80
1.00
1.20
1.40
1.60
Minutes
34
1.80
2.00
Recommendations Parameters:
System Suitability
Capacity factor
k' > 2
Precision/Injection repeatability
RSD </= 1%, n >/= 5
Resolution
Rs >/= 2 (Major peak and
closest eluting)
Tailing factor
T </= 2
Theoretical Plates
In general N > 2000
35
INJ VOL
No of Inj
Function
Method
Blank
20.0
Inject Samples
System
Suitability
20.0
Inject Samples
Std1
20.0
Clear
Calibration
5 Inject Standards
3
4
Std2
20.0
1 Inject Standards
Dilution
10.00
1.00000
1.00000
10.00
1.00000
1.00000
10.00
1.00000
1.00000
10.00
1.00000
1.00000
Report
LC Calibration Report
Report
Standard Comparison
Std1
20.0
Clear
Calibration
1 Inject Standards
10.00
1.00000
1.00000
Unk.1
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.2
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.3
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.4
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.5
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.6
20.0
Inject Samples
10.00
1.00000
1.00000
Std1
20.0
1 Inject Standards
10.00
1.00000
1.00000
1
2
Clear
Calibration
Calibrate
Summary Report
Sample Set: PrednisoneNoRecircNoRepl
General Information
Sample Set Name
Sample Set Method
Sample Set Start Date
Sample Set Finish Date
Sample Set Id
Sample Set Altered
SampleName
PrednisoneNoRecircNoRepl
PrednisoneNoRecircNoRepl
27/10/1999 12:36:43
27/10/1999 15:37:29
2090
No
Sample Type
Vial
Run Time
(Minutes)
Inj #
Injection
Volume
(ul)
Acquistion
Method Set
Sample
Weight
Dilution
Label
Prednisone_Tab
Unknown
11
4.00
20.00
1.00000
1.00000
AU0203
Prednisone_Tab
Unknown
12
4.00
20.00
1.00000
1.00000
AU0204
Prednisone_Tab
Unknown
14
4.00
20.00
1.00000
1.00000
AU0206
Prednisone_Tab
Unknown
16
4.00
20.00
1.00000
1.00000
AU0208
Area
Calibration Plot
1.0x10
5.0x10
0.0
0.000
0.002
0.004
0.006
0.008
0.010
Amount
0.012
0.014
Name: Prednisone; Processing Method: Prednisone Proc Mth; Fit Type: Linear (1st
Order); Cal Curve Id: 2973; A: 0.000000e+000; B: 4.867524e+007; C:
0.000000e+000; D: 0.000000e+000; R^2: 1.000000
36
0.016
0.018
0.020
Assay
Calibration Plot
OR:
Area
6
1.0x10
5
5.0x10
0.0
0.0000.0020.0040.0060.0080.0100.0120.0140.0160.0180.020
Related Compounds
MAIN
37
= W Std/AStd
Working Curve
A plot
of
response)
the
analytical
as a function of
signal
(the
analyte
concentration,
instrument
or
detector
using a series
sample or to
calibrate
the concentration
of
instrument.
Choice of Standardization:
External or Internal
External Standard
Response Std
Amount Std
Amount Unk
Response Unk
Amount Std
Amount Unk =
Response Std
Response
Std
Amount
Std
38
Response Unk
Choice of Standardization
External or Internal
Simple formulations and sample
preparation: external standard
Choice of Standardization:
External or Internal
Internal Standard
C
Int. Std.
Amount Std
Amount Istd
Response Std
Response Istd
Amount Unk
Response Unk
Response Istd
Amount Istd
39
Internal Standard
12,000/6000
11,000/5800
100/10
Amt B/10
B
Int.
Std.
Int.
Std.
100
10
11,000
10
12,000
5,800
Unknown
6,000
Standard
(11,000/5,800) x (100/10)
----------------------------------- x 10 = Amt B =
12,000/6000
94.8
Standard Addition
Due to matrix effects the analytical response for an
analyte in a complex sample may not be the same as
for the analyte in a simple standard.
Signal
Conc.
40
Summary Report
Sample Set: PrednisoneNoRecircNoRepl
General Information
Sample Set Name
Sample Set Method
Sample Set Start Date
Sample Set Finish Date
Sample Set Id
Sample Set Altered
SampleName
PrednisoneNoRecircNoRepl
PrednisoneNoRecircNoRepl
27/10/1999 12:36:43
27/10/1999 15:37:29
2090
No
Sample Type
Vial
Run Time
(Minutes)
Inj #
Injection
Volume
(ul)
Acquistion
Method Set
Sample
Weight
Dilution
Label
Prednisone_Tab
Unknown
11
4.00
20.00
1.00000
1.00000
AU0203
Prednisone_Tab
Unknown
12
4.00
20.00
1.00000
1.00000
AU0204
Prednisone_Tab
Unknown
14
4.00
20.00
1.00000
1.00000
AU0206
Prednisone_Tab
Unknown
16
4.00
20.00
1.00000
1.00000
AU0208
Area
Calibration Plot
1.0x10
5.0x10
0.0
0.000
0.002
0.004
0.006
0.008
0.010
0.012
0.014
0.016
0.018
0.020
Amount
Name: Prednisone; Processing Method: Prednisone Proc Mth; Fit Type: Linear (1st
Order); Cal Curve Id: 2973; A: 0.000000e+000; B: 4.867524e+007; C:
0.000000e+000; D: 0.000000e+000; R^2: 1.000000
Standard Deviation
A measure of the uncertainty due to random
error in a set of data (also: precision of a set of
measurements).
s
N
x
x
= standard deviation,
= the num ber of data points,
i = each individual m easurem ent,
bar = the mean of all m easurem ents.
41
SAMPLE NAME
INJ VOL
No of Inj
Function
Blank
20.0
Inject Samples
2 System
Suitability
20.0
Inject Samples
Std1
20.0
Clear
Calibration
5 Inject Standards
Std2
20.0
1 Inject Standards
Method
Run Time
Sample
Weight
1.00000
Dilution
10.00
10.00
1.00000
1.00000
10.00
1.00000
1.00000
10.00
1.00000
1.00000
1.00000
Report
LC Calibration Report
Report
Standard Comparison
Std1
20.0
Clear
Calibration
1 Inject Standards
10.00
1.00000
1.00000
Unk.1
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.2
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.3
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.4
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.5
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.6
20.0
Inject Samples
10.00
1.00000
1.00000
Std1
20.0
1 Inject Standards
10.00
1.00000
1.00000
1
2
Clear
Calibration
Calibrate
Control Std2
Std2 (Accuracy/Recovery Test)
42
Standards_Comparison
1
2
3
4
5
6
Mean
% RSD
SampleName
std-1
std-1
std-1
std-1
std-1
std-1
Label
S1
S1
S1
S1
S1
S1
Name
methyl paraben
methyl paraben
methyl paraben
methyl paraben
methyl paraben
methyl paraben
Vial Inj
RT
2
1 2.775
2
2 2.775
2
3 2.777
2
4 2.775
2
5 2.773
2
6 2.775
Area
7928853
7926182
7973699
7993516
7993709
8004153
Amount Area_Amount
44.429
178461
44.429
178401
44.429
179471
44.429
179917
44.429
179921
44.429
180156
179388
0.43
1
2
Mean
SampleName Label
std-2
S2
std-2
S2
Name
methyl paraben
methyl paraben
Vial Inj
RT
1
1 2.783
1
2 2.779
Area
7858421
7907563
SampleName
std-1
std-1
std-1
std-1
std-1
std-1
Label
S1
S1
S1
S1
S1
S1
Name
propyl paraben
propyl paraben
propyl paraben
propyl paraben
propyl paraben
propyl paraben
Vial Inj
RT
2
1 9.067
2
2 9.068
2
3 9.066
2
4 9.064
2
5 9.053
2
6 9.058
Area
670840
670683
673923
678792
676660
678451
Amount Area_Amount
45.014
14903
45.014
14899
45.014
14971
45.014
15080
45.014
15032
45.014
15072
14993
0.54
1
2
Mean
SampleName Label
std-2
S2
std-2
S2
Name
propyl paraben
propyl paraben
Vial Inj
RT
1
1 9.098
1
2 9.075
Area
670235
673646
Project Name:
Control_Check
100.72
100.21
100.5
Pharma_QC_calculations_Trima
Date Printed:
16/04/2010
21:02:38 Asia/Tel_Aviv
Test of Accuracy
Response Std
Amount Std
Amount Control
Response Unk
Amount Std
Amount Control =
Response Std
x Response Unk
=0.99-1.01
Accuracy:
Amount Control Prepared
43
Running the
Unknowns
No of Inj
Function
Method
Dilution
Blank
20.0
Inject Samples
10.00
1.00000
1.00000
2 System
Suitability
20.0
Inject Samples
10.00
1.00000
1.00000
10.00
1.00000
1.00000
10.00
1.00000
1.00000
Std1
20.0
Clear
Calibration
5 Inject Standards
Std2
20.0
1 Inject Standards
Report
LC Calibration Report
Report
Standard Comparison
Clear
Calibration
Unk.1
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.2
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.3
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.4
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.5
20.0
Inject Samples
10.00
1.00000
1.00000
Unk.6
20.0
Inject Samples
10.00
1.00000
1.00000
Std1
20.0
1 Inject Standards
10.00
1.00000
1.00000
Clear
Calibration
Calibrate
44
Basic Report
Reported by User:
System
Project Name:
SAM PLE
Sample Name:
Sample Type:
Vial:
Injection #:
Injection Volume:
Run Time:
Sample Set Name:
Untitled
Prednisone_Tab
Unknown
16
1
20.00 ul
4.0 Minutes
PrednisoneNoRecircNoRepl
Dissolution_Default
IN FO R MATION
Acquired By:
Date Acquired:
Acq. Method Set:
Date Processed:
Processing Method:
Channel Name:
Proc. Chnl. Descr.:
System
27/10/1999 14:38:40
Prednisone Mth Set
27/03/2001 14:17:26
Prednisone Proc Mth
2487Channel 1
Single @ 242 nm
Auto-Scaled Chromatogram
Prednisone - 1.791
0.18
0.16
0.14
AU
0.12
0.10
0.08
0.04
0.02
0.573
0.06
0.00
0.50
1.00
1.50
2.00
Minutes
2.50
3.00
3.50
4.00
Peak Results
Name
1
2
Prednisone
RT
Area
Height
0.573
22575
822
1.791
1188824
186546
Amount
Units
0.024
mg/ml
Calibration Procedures
Calibration
Standards at
the beginning
only
Calibration
Standards at the
beginning and
end
Calibration
Standards
Bracketing
45
Calibration
Standards
Bracketting
Calibration
Standards
Bracketting
Calibration
Standards
Bracketting
46
Linearity
Day 1-3
LOQ
LOD
Parameter 1
Min & Max
Level 1: 60%
Bulk & Formul.
Level 1
Bulk Drug
Six Blanks
Baseline Noise
Parameter 2
Min & Max
Level 2: 80%
Bulk& Formul.
Level 2
Bulk Drug
Parameter 3
Min & Max
Level 3: 100%
Bulk & Formul.
Level 3
Bulk Drug
Parameter 4
Min & Max
Level 4: 120%
Bulk & Formul.
Level 4
Bulk Drug
Level 5: 140%
Bulk & Formul.
Level 5
Bulk Drug
LOQ:
Precision
Precision
Day 1-3
Six Reps., 100%
Bulk Drug
LOQ Level
Bulk Drug
47
Precision:
The reproducibility of results. The degree to which an
experimental result varies from one determination to
next.
the
Precision: Definition
Precision
The measure of the degree of agreement among test results when the method
is applied repeatedly to multiple samplings of a homogeneous sample
Expressed as %RSD for a statistically significant number of samples
48
Precision: Definition
Precision Should Be Performed at Three Levels
Repeatability
Intermediate Precision
Reproducibility
Precision: Definition/Determination
Repeatability (Generally the criterion of concern in USP analytical procedures)
Same operating conditions, short time interval
Intra-assay precision
Minimum of 9 determinations covering specified range of procedure (3
levels, 3 reps each), or
Minimum of 6 determinations at 100% test conc.
Reproducibility
Precision between labs
Collaborative studies
49
Ruggedness: Definition
Ruggedness
Degree of Reproducibility of Test Results Under a Variety of
Conditions
Different Laboratories
Different Analysts
Different Instruments
Different Reagents
Different Days
Etc.
Expressed as %RSD
Calculating Precision
N
X
Mean :
Standard Deviation :
Coefficient
of Variation :
CV% =
N 1
50
(100 )
Response
125955
125651
126052
124855
126102
125899
Average
125752
Standard Deviation
466.93
0.37%
Precision Test
0.25
0.20
AU
0.15
0.10
0.05
0.00
1.00
2.00
SampleName: SysSuit_A;
SampleName: SysSuit_A;
SampleName: SysSuit_A;
SampleName: SysSuit_A;
SampleName: SysSuit_A;
SampleName: SysSuit_A;
3.00
Vial: 5;
Vial: 5;
Vial: 5;
Vial: 5;
Vial: 5;
Vial: 5;
Injection: 1;
Injection: 2;
Injection: 3;
Injection: 4;
Injection: 5;
Injection: 6;
4.00
5.00
Minutes
Channel: 2487Channel 1;
Channel: 2487Channel 1;
Channel: 2487Channel 1;
Channel: 2487Channel 1;
Channel: 2487Channel 1;
Channel: 2487Channel 1;
6.00
7.00
8.00
9.00
SampleName Inj
Channel
Name
SysSuit_A
1 2487Channel 1 EtP
SysSuit_A
2 2487Channel 1 EtP
SysSuit_A
3 2487Channel 1 EtP
SysSuit_A
4 2487Channel 1 EtP
SysSuit_A
5 2487Channel 1 EtP
SysSuit_A
6 2487Channel 1 EtP
RT
3.997
4.002
3.978
3.993
3.979
4.010
Area
1659138
1657741
1659683
1659981
1660539
1657722
1659134
1177
0.1
51
10.00
%RSD
0.40
0.30
0.20
0.10
0.00
Area
Height
Retention
Time
Resolution
52
Accuracy:
The degree to which an experimental result
approaches the true or accepted answer.
Ways to Describe Accuracy:
Error:
ERRORS
Random Error: results in a scatter of results centered on the true value
for repeated measurements on a single sample.
(SPREAD)
Systematic Error: results in all measurements exhibiting a definite
difference from the true value (BIAS)
Random Error
Systematic Error
53
SD =
(X X )
i
/( n 1)
i =1
Overlaid Chromatograms
Good Precision
0.05
0.04
0.03
AU
Peak1
0.02
Peak2
Peak3
0.01
0.00
0.00
1.00
2.00
3.00
4.00
5.00
6.00
Minutes
SampleName 2690_Eau40_60CH3OH_2mm_D Vial 4 Injection 1
SampleName 2690_Eau40_60CH3OH_2mm_D Vial 4 Injection 2
SampleName 2690_Eau40_60CH3OH_2mm_D Vial 4 Injection 3
SampleName 2690_Eau40_60CH3OH_2mm_D Vial 4 Injection 4
SampleName 2690_Eau40_60CH3OH_2mm_D Vial 4 Injection 5
SampleName 2690_Eau40_60CH3OH_2mm_D Vial 4 Injection 6
54
7.00
8.00
Precision (Ruggedness)
M e th o d s Tra n s fe r: 3 H P L C S ys te m s
Accuracy
3 S yste m s
1 in U S A
2 in E u ro p e
3 C h e m is ts
D ia l-a -m ix , 6 0 :4 0
M e O H :H 2 O
1 m L /m in
3 0C
0.05 0
0.04 0
Limit of detection
Limit of quantitation
0.03 0
AU
0.02 0
Linearity (range)
0.01 0
Selectivity
0.00 0
1 .0
2.0
3 .0
4.0
5 .0
6.0
7 .0
Robustness
M in u te s
0.050
0.040
0.030
AU
0.020
0.010
0.000
1.0
2.0
3.0
4.0
Minutes
55
5.0
6.0
7.0
Column BatchBatch-to
to--Batch
Reproducibility
0.02
Imp1
Imp2
AU
Imp3
Batch 108
Injection: 150
Imp4
0.00
Sample: AZT
L of 0.5 mg/mL
solution
10
0.02
Batch 109
Imp1
Imp2
AU
20
Minutes
Imp3
Imp4
0.00
THF/ 88%
10
0.02
20
Minutes
10 mM potassium phosphate
buffer, pH 2.5
Imp1
Batch 112
Imp3
AU
Imp2
Detector: UV at 268 nm
Imp4
0.00
10
20
Minutes
Integration parameters
Injector performance
Pump performance
56
- Detector
* Cell Problem
* Lamp Failing
AU
0.005
0.000
51.8 52.0 52.2 52.4 52.6 52.8
Minutes
Injector Linearity
Response
2,000
1,500
Different
volumes of
the same
standard
1,000
Methods
transfer
R2 = 0.9999
500
10
20
30
40
Injection Volume L
57
50
Accuracy: Definition
The closeness of test results obtained by the method to the true
value.
Established across the concentrations range
Accuracy: Determination
BioBio-active Substance
Analysis of reference material
Bio
Bio--active Formulation
Analysis of synthetic mixtures spiked with known quantities
of components
Impurities (Quantitation)
Analysis of samples (Bio-active substances/Bio-active
formulation) spiked with known amounts of impurities
If impurities are not available, see specificity
58
Recommended Data
Minimum of 9 determinations over a minimum of 3
concentration levels covering the specified range (e.g. 3
concentrations/3 replicates each)
Reported as % recovery of known, added amount, or
difference between the mean and true value, with
confidence intervals
Accuracy of Measurement
Correlation Coefficient
(Xi
) (Yi Y )
(X X )2 (Y Y )2
X
(X
i,
Yi )
Error :
% Relative Error :
E = O - T
%RE =
E
T
59
x 100
O = Observed Value
T = True Value
Spiked Concentration
(mg/mL)
Measured
Concentration
(mg/mL) Avg N=3
% Recovery
50%
0.50
0.49
98.0
75%
0.75
0.74
98.7
100%
1.00
1.03
103
125%
1.25
1.24
99.2
150%
1.50
1.48
98.7
Average Recovery:
Standard Deviation:
Relative Standard Deviation:
99.52%
1.99
2.00
Documenting Accuracy
with Control Charts
Custom Report Control Chart for: ACE, Channel: 996
411879.8
401879.8
391879.8
Area
381879.8
371879.8
UCL
361879.8
Target
351879.8
LCL
341879.8
331879.8
321879.8
1
Run Number
60
10
11
12
h signal = 2 x h noise
Precision (Ruggedness)
h signal
Accuracy
h noise
Limit of detection
Limit of quantitation
LOQ - RSD < 5-10%
Linearity (range)
0.010
Selectivity
AU
0.005
Robustness
0.000
51.8 52.0 52.2 52.4 52.6 52.8
Minutes
DL = (3.3)STD/S
STD = standard deviation of the response
S = slope of the calibration curve
61
QL = (10)STD/S
STD = standard deviation of the response
S = slope of the calibration curve
Both DL and QL are validated by analyzing a suitable number
of samples. Method should be documented
62
Level 2 Response
Level 3 Response
Level 4 Response
Level 5 Response
2085
4105
6288
8333
10499
2106
4235
6103
8287
10305
1955
4188
6337
8399
10386
2199
4063
6129
8156
10355
2077
4005
6254
8247
10462
2155
4284
6301
8178
10219
Mean
2096
4147
6235
8267
10371
STDev.
83.13
106.9
96.53
92.54
102.57
96.33
9378
0.034 mg/mL
0.103 mg/mL
A) Brand Y C18
Signal to Noise = 11
AUFS
5.00
Minutes
10.00
0.0005
B) Brand X C18
Signal to Noise = 6.5
AUFS
5.00
Minutes
10.00
63
Precision (Ruggedness)
3.5
3
Accuracy
Absorbance (AU)
2.5
2
Limit of detection
1.5
1
Limit of quantitation
0.5
0
0
10
20
30
40
50
60
70
80
% Absorbe r in Elue nt
Linearity (range)
Selectivity
Robustness
64
Linearity: Determination
Established across the Range of the method
Dilutions
Separate Weighings
Evaluate by Appropriate Statistical Methods (e.g. Regression)
Include Correlation Coefficient, y-Intercept, Slope, Residual
Sum of Squares, Plot Itself
Minimum 5 Concentrations
65
Response
5,000,000
4,000,000
3,000,000
2,000,000
1,000,000
0
0.02
0.03
0.04
0.05
0.06
0.07
0.08
0.09
0.10
Amount
R-square = 1 # pts = 5
y = -8.95e+005 + 5.4e+007x
66
0.11
%RSD-Time
HCT
%RSD-Time
TMT
%RSD-Area
HCT
%RSD-Area
TMT
1
2
3
0.00
0.05
0.00
0.00
0.00
0.00
0.05
0.16
0.18
0.06
0.07
0.05
Intermediate Precision:
N = 18 (6 inj./day, 3 days) at 100%
Compound
%RSD-Time
%RSD-Area
HCT
0.21
3.24
TMT
0.56
3.97
200.00
Precision (Ruggedness)
nm
0.40
0.20
Maximum
Impurity
Accuracy
AU
0.00
2.20
2.40
2.60
2.80
Limit of detection
3.00
Minutes
Limit of quantitation
Photodiode Array Technology
Linearity (range)
Spectral Analyses
Library Matching
Compound identification
Coelution detection
Selectivity/Specificity
Robustness
67
Specificity: Definition
Specificity (Selectivity)
The ability to measure accurately and specifically the analyte in
the presence of components that may be expected to be present
in the matrix
The degree of interference
Active Ingredients
Excipients
Impurities
Degradation Products
Placebo Ingredients
Specificity: Determination
Qualitative Identification Tests
Demonstrate ability to select between compounds of closely
related structure
Assay
Demonstrate that the results are unaffected by spiked impurities
or excipients
Impurities
Spike the drug product/substance with impurities and
demonstrate appropriate accuracy and precision
68
Impurities
Degradants
Composite sample
Courtesy of:
Dr. Rudy Sneyers, J&J Belgium
69
70
When to Use
Stability Indicating Methods
When are SIMs needed?
Stability studies
API release
Drug product release
Toxicology dosing solutions
Excipient compatibility and pre-formulation
Packaging studies
Line extensions
Inorganics
Sample Generation
Forced Degradation (Stress) Studies
Forced degradation or stress testing is undertaken to demonstrate
specificity when developing SIMs
Generates a sample for method development
71
Sample Generation
Common conditions used in Forced Degradation Studies
Study
Conditions
Acidic pH
0.1N HCl
Neutral pH
Basic pH
0.1N NaOH
Oxidation
02 Atmosphere, or H2O2
Photolysis (UV)
Photolysis (Fluorescence)
6x106 lux h
Stability Indicating
Method Development
Manipulate chromatographic selectivity
Column
Mobile phase composition/type
pH
Temperature
Specificity
PDA or MS
Goal:
72
+
PDA and MS together can
provide complimentary useful
information.
73
1.20
1.25
1.30
1.35
1.40
1.45
1.50
1.55
1.60
Minutes
74
1.65
1.70
1.75
1.80
1.85
AU
0.14
ION DETECTOR
+
+
SOURCE
+
+
+
+
DETECTION OF IONS
+
+
SORTING OF IONS
+
+
SAMPLE DESOLVATION
AND IONIZATION
LC/MS
INTERFACE
HPLC
DATA SYSTEM
MASS SPECTRUM
CHROMATOGRAM
6.000e+007
5.000e+007
4.000e+007
3.000e+007
2.000e+007
1.000e+007
0.000e+000
200.00
400.00
600.00
800.00
1000.00
1.00
1.05
1.10
1.15
1.20
1.25
1.30
1.35
1.40
1.45
1.50
1.55
Minutes
75
1.60
1.65
1.70
1.75
1.80
m/z
Intensity
7.000e+007
250.00
nm
300.00
nm
300.00
250.00
350.00
1.19
Peak1
0.25
250.00
350.00
nm
300.00
350.00
1.32
Peak 2
1.19
1.75
Peak 3
1.31
1.74
1.20
1.33
1.76
1.18
1.31
1.74
1.21
1.34
1.77
Peak1
Peak 2
Peak 3
AU
0.20
0.15
0.10
0.05
0.00
1.00
1.10
1.20
1.30
1.40
1.50
Minutes
1.60
1.70
1.80
1.90
2.00
Chromatogram of three sulfa drugs at 270 nm and a plot of UV Spectra collected from each peak's Apex, Inflection and two
offset points, to demonstrate comparison of spectra collected at various points across the peak.
MS Spectra
1.2x107
265.1
8.0x106
1.0x107
Peak 1
6.0x106
5.0x106
4.0x106
2.0x106
5x107
Intensity
Intensity
7
6x10
279.1
4x107
3x107
Peak 2
2x107
529.2
530.1
279.1
2x107
Peak 2
280.2
281.1
557.2
8x107
8x107
311.1
6x107
4x107
Intensity
Intensity
266.1
267.1
4x107
1x107
Peak 3
1.20
1.30
1.40
1.50
1.60
1.70
1.80
311.1
6x107
4x107
2x107
2x107
Peak 1
0.0
0.0
1.10
265.1
Intensity
Intensity
1.0x107
1.5x107
Peak 3
312.2
313.1
0
200.00
300.00
400.00
500.00
m/z
Minutes
SIR chromatograms of the same sulfa drugs and their corresponding mass spectra, obtained by combining
spectra across the entire peak.
76
600.00
N
NH
O
0.85
0.80
Lansoprazole
0.75
0.70
UV @ 254nm
O
F
0.65
0.60
0.55
-0.00125
0.50
-0.00130
UV @ 254nm
254nm
0.45
0.03%
AU
-0.00135
0.40
-0.00140
0.35
S/N = 2
-0.00145
0.30
-0.00150
AU
0.25
0.20
0.15
-0.00155
-0.00160
0.10
-0.00165
0.05
-0.00170
0.00
-0.00175
-0.05
0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00
Minutes
-0.00180
4.80
5.00
5.20
5.40
Minutes
5.60
5.80
6.00
N
NH
O
N
0.85
0.80
0.75
0.70
0.65
Lansoprazole
O
UV @ 254nm
SIR @ 298.2 m/z
F
F
0.60
0.55
2.0x106
0.50
1.8x106
AU
0.45
0.40
1.6x106
0.35
1.4x106
S/N = 870
Intensity
0.30
0.25
0.20
1.2x106
1.0x106
0.15
8.0x105
0.10
6.0x105
0.05
4.0x105
0.00
2.0x105
-0.05
0.50
1.00 1.50
2.00 2.50
3.00 3.50
4.00
6.50 7.00
7.50 8.00
8.50 9.00
9.50 10.00
0.0
4.80
77
5.00
5.20
5.40
5.60
Minutes
5.80
6.00
Sample Generation
Common conditions used in Forced Degradation Studies
Study
Conditions
Acidic pH
0.1N HCl
Neutral pH
Basic pH
0.1N NaOH
Oxidation
02 Atmosphere, or H2O2
Photolysis (UV)
Photolysis (Fluorescence)
6x106 lux h
Risperidone
0.06
0.04
0.02
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
11.00
12.00
Minutes
0.12
Risperidone
0.10
AU
0.08
80%A: 20%B
0.06
0.04
0.02
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
11.00
12.00
Minutes
0.12
Risperidone
0.10
AU
0.08
Conditions:
Column Dimensions: 4.6 x 100 mm
Isocratic separation
A: pH 3.0; 10mM NH4COOH in H2O
B: Acetonitrile
Wavelength: 280nm
Flow rate: 1.4 mL/min
Injection volume: 10.0 L
Temperature: 30 oC
Risperidone concentration:
0.5 mg/ ml in 1.55% H2O2
Risperidone degraded ~ 15 - 30%
82%A: 18%B
0.06
Instrumentation:
Waters Alliance HT 2790
Waters 996 PDA detector
0.04
0.02
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
11.00
12.00
0.12
Risperidone
0.10
0.08
AU
AU
0.08
0.06
0.04
0.02
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
Minutes
78
10.00
11.00
12.00
UVUV-VIS
79
Precision (Ruggedness)
Accuracy
5N-2,4,6T
4A-6C-1,3B
K'
HCT
Limit of detection
TMT
Limit of quantitation
0
10
30 50 70 90
20 40 60 80 100
Linearity (range)
mM NaH2PO4 pH 5.5
Selectivity
Robustness
80
Robustness: Definition
Robustness
Measure of The Capacity to Remain Unaffected by Small
(Deliberate) Variations in Method Parameters
Indication of Reliability During Normal Use
Robustness: Determination
Consider during development of method
Shows reliability of method with respect to deliberate changes
If measurements are susceptible to variations in analytical
procedures, these conditions should be controlled and a
precautionary statement included.
Establish System Suitability parameters to ensure the validity of the
method
81
:
Solvent strength in the mobile phase,
Temperature,
Flow rate,
pH of the mobile phase,
Ionic strength in the mobile phase,
Sample diluent,
Injection volume,
Wavelength of detection.
Non--Robust Method
Non
pH=4.5
pH=4.9
Minor Changes in
pH Effect Rs
pH=4.7
82
Robust Method
AZT: Robustness Testing
6% M ethanol, 6% THF
AU
0.010
Imp. 1
pH 2.3
Imp. 3
0.008
Imp. 2
0.006
0.004
Imp. 4
0.002
0.000
-0.002
0
10
15
20
Imp. 1
AU
0.010
Imp. 3
0.008
pH 2.5
Imp. 2
0.006
0.004
Imp. 4
0.002
0.000
-0.002
0
10
15
20
Time [min]
AU
Im p. 1
0.010
Imp. 3
0.008
pH 2.7
Imp. 2
0.006
0.004
Imp. 4
0.002
0.000
-0.002
0
10
20
15
Time [min]
83
84
Login.
Instrument Method.
Sequence/Run Samples.
Process Data.
Review/Preview.
Result/Sign-Off.
3.
2.
4.
5.
6.
1.
85
86
System
No
3767
4
-1.0
5.0
0.0
0.0
0.0
0.0
-1.0
5.0
2.00
No Change
0.00
Normal
No Change
No Change
No Change
No Change
False
Detector Information
Channel Name
Description
Use Channels
2487Channel1
On
Wavelength
254
Output Mode
Data Mode
Sampling Rate
Filter Type
Absorbance A(Ch1)
Absorbance A(Ch1)
2
Hamming
Aufs
2.0000
Time Constant
1.0
AU Offset
0.000
Voltage Offset
0
Polarity
+
AutoZero Wavelength True
AutoZero Keypad
True
AutoZeroEvent Input True
87
AutoZero Inject
Chart Mark Enable
Ratio AuMinimum
Minimum Ratio
Maximum Ratio
True
True
0.1000
0.00
2.00
Basic Report
Reported by User:
System
Project Name:
SAM PLE
Sample Name:
Sample Type:
Vial:
Injection #:
Injection Volume:
Run Time:
Sample Set Name:
Untitled
Prednisone_Tab
Unknown
16
1
20.00 ul
4.0 Minutes
PrednisoneNoRecircNoRepl
Dissolution_Default
IN FO R MATION
Acquired By:
Date Acquired:
Acq. Method Set:
Date Processed:
Processing Method:
Channel Name:
Proc. Chnl. Descr.:
System
27/10/1999 14:38:40
Prednisone Mth Set
27/03/2001 14:17:26
Prednisone Proc Mth
2487Channel 1
Single @ 242 nm
Auto-Scaled Chromatogram
Prednisone - 1.791
0.18
0.16
0.14
AU
0.12
0.10
0.08
0.04
0.02
0.573
0.06
0.00
0.50
1.00
1.50
2.00
Minutes
2.50
3.00
Peak Results
Name
1
2
Prednisone
RT
Area
Height
0.573
22575
822
1.791
1188824
186546
Amount
0.024
Units
mg/ml
Processing Method
Method Set
Instrumental parameters
Processing parameters
Reporting
Exporting
88
3.50
4.00
89
Processing Method
Type : LC
: Parabens Ten
Processing Method
Parameters: Example
Method Information
Method Comments
Method Modified User
System
Method Locked
No
Method Id
2193
Method Version
12
Method Edit User
Source S/W Info
Beta Empower Software Build
1154 DB ID: 685141106
Integration Algorithm
ApexTrack
Average By
None
RT Window %
5.00
Update RT
Never
CCalRef1
Include Internal Standard Amounts in Amount Calculation
Yes
Vial/Default Value Type
Amount
System Suitability Information
Void Volume Time
1.000(min)
Calculate Suitability
Yes
Flag Outside
Yes
Calculate Unknowns
Yes
Pharmacopoeia
All
Tangent Percent (USP Plate Count)61
Integration Parameters
Minimum Area
0(V*sec)
Minimum Height 0(V)
Integration Start 2.000(min)
Integration End
(min)
Peak Width
11.42(sec)
Detection Threshold 5.000e+002(V)
Liftoff %
Touchdown %
Baseline Start
Baseline End
(min)
(min)
0.000(%)
0.500(%)
Report Method
Method Set
Instrumental parameters
Processing parameters
Reporting
Exporting
90
Basic Report
Reported by User:
System
Project Name:
SAM PLE
Sample Name:
Sample Type:
Vial:
Injection #:
Injection Volume:
Run Time:
Sample Set Name:
Untitled
Prednisone_Tab
Unknown
16
1
20.00 ul
4.0 Minutes
PrednisoneNoRecircNoRepl
Dissolution_Default
IN FO R MATION
Acquired By:
Date Acquired:
Acq. Method Set:
Date Processed:
Processing Method:
Channel Name:
Proc. Chnl. Descr.:
System
27/10/1999 14:38:40
Prednisone Mth Set
27/03/2001 14:17:26
Prednisone Proc Mth
2487Channel 1
Single @ 242 nm
Auto-Scaled Chromatogram
Prednisone - 1.791
0.18
0.16
0.14
AU
0.12
0.10
0.08
0.06
0.573
0.04
0.02
0.00
0.50
1.00
1.50
2.00
Minutes
2.50
3.00
3.50
4.00
Peak Results
Name
1
2
Prednisone
RT
Area
Height
0.573
22575
822
1.791
1188824
186546
Amount
0.024
Units
mg/ml
Summary Report
Sample Set: PrednisoneNoRecircNoRepl
General Information
Sample Set Name
Sample Set Method
Sample Set Start Date
Sample Set Finish Date
Sample Set Id
Sample Set Altered
SampleName
PrednisoneNoRecircNoRepl
PrednisoneNoRecircNoRepl
27/10/1999 12:36:43
27/10/1999 15:37:29
2090
No
Sample Type
Vial
Run Time
(Minutes)
Inj #
Injection
Volume
(ul)
Acquistion
Method Set
Sample
Weight
Dilution
Label
Prednisone_Tab
Unknown
11
4.00
20.00
1.00000
1.00000
AU0203
Prednisone_Tab
Unknown
12
4.00
20.00
1.00000
1.00000
AU0204
Prednisone_Tab
Unknown
14
4.00
20.00
1.00000
1.00000
AU0206
Prednisone_Tab
Unknown
16
4.00
20.00
1.00000
1.00000
AU0208
Area
Calibration Plot
1.0x10
5.0x10
0.0
0.000
0.002
0.004
0.006
0.008
0.010
Amount
0.012
0.014
Name: Prednisone; Processing Method: Prednisone Proc Mth; Fit Type: Linear (1st
Order); Cal Curve Id: 2973; A: 0.000000e+000; B: 4.867524e+007; C:
0.000000e+000; D: 0.000000e+000; R^2: 1.000000
91
0.016
0.018
0.020
Quantitative Results
SAM PLE
I N F O R MATI O N
Sample Name :
Sample Type :
Vial :
Injection #:
Injection Volume :
Run Time :
K -26525 /1-H 2O 2
Unknown
24
1
10 .00 ul
55 .0 Minutes
Acquired By :
Sample Set Name :
Acq . Method Set :
Processing Method :
Channel Name :
Proc . Chnl . Descr .:
Date Acquired :
Date Processed :
Sofia
PDA Report:
Example
Deg1 - 8.611
200.00
nm
250.00
Main - 11.663
200.00
nm
300.00
250.00
193.7
250.00
193.7
6.74
6.69
6.81
nm
300.00
300.00
193.7
8.61
8.54
8.71
11.66
11.56
11.84
Main - 11.663
Deg1 - 8.611
0.50
6.737
AU
1.00
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
11.00
12.00
13.00
14.00
Minutes
1
2
3
Name
RT
Deg1
Main
6.737
8.611
11.663
Purity1
Angle
1.854
0.261
0.041
Purity1
Threshold
2.483
0.439
0.240
Purity1
Flag
No
No
No
92
Match1
Angle
1.732
4.183
0.024
Match 1
Threshold
2.489
1.142
1.008
PDA/FLR Match 1
Flag
No
Yes
No
15.00
Match Plot
Match Plot
193.7
Deg1
Match #1 Angle 4.183 Main 100%
288.6
Main
Match #1 Angle 0.024 Main 100%
288.6 288.6
291.0
80.00
Deg1 - 8.611
Purity
Auto Threshold
40.00
30.00
60.00
40.00
AU
0.02
1.00
Degrees
AU
Purity Plot
Purity
Auto Threshold
Main - 11.663
Purity Plot
0.04
0.50
20.00
10.00
20.00
0.00
0.00
8.40
8.60
8.80
PDA Report:
Example
9.00
0.00
0.00
11.40
Minutes
11.60
11.80
12.00
Minutes
12.20
12.40
12.60
Reports to Connect
Data to Decisions
93
Degrees
193.7
Electronic Signatures
Chromatographic
Data System (CDS)
Additional
Calculations
Performed
Reports Compiled
LIMS
Final Review
Signatures Applied
94
Reports Reviewed
Empower
CDS
Additional
Calculations
Performed
Reports Compiled
Reports Reviewed
Signatures Applied
95
Final Review
96
97