Asia Pesticide Residue Mitigation through the Promotion of Biopesticides and Enhancement of Trade Opportunities (APRMP), Virtual lab meeting 13 August 2020

1. GLP TRAINING
 Wayne Jiang, PhD
IR-4, Michigan State University, jiangwa@msu.edu
 Michael Braverman, PhD
IR-4 Headquarters, Rutgers University
Extraction and Analysis

2. Review
• Sample shipping, Sample:dry ice ratio, in
container, separate control and treated, temp monitoring
• Sample check, receipt, unique ID
• Short-term and long-term storage
• Temperature monitoring, alarm, phone
• Types of grinders, grinding with dry ice
• Sample homogeneity and integrity
• GLP Reference substances
• Recertification of GLP standards
• Labels (required information)
• Critical phase inspection

3. True or False
A copy of an original raw data page directly
from a Canon photocopier is an exact copy of
the raw data.
A computer printout derived from data
transferred to computer media from laboratory
data sheets is considered to be raw data.

4. True or False
GLP studies may be done in facilities that
utilize other quality standards (GMP, GCP, ISO
etc.).
EPA inspector can take photographs or
photocopies of objectionable practices and
conditions.

5. Q & A
Raw data must be co-signed by a second
individual.
Raw data must be directly entered into bound
notebooks.

6. Q & A
QAU must keep a copy of lab SOPs.
Study director authorizes changes in the SOPs.

7. Q & A
What are the GLP requirements for labeling of
solvents purchased directly from
manufacturers?
(select all correct answers)
(A) Expiration date
(B) Identity
(C) User’s initial and date
(D) Safety information
(E) Storage condition.

8. Q & A
Which role can a study director be ?
(Select all correct answers.)
(A) Archivist
(B) Field Research Director
(C) Lab analyst
(D) Company CEO
(E) QA

9. Definitions
• Limit of Detection (LOD) is the smallest amount
of the analyte that can be reliably detected from
the background for a particular matrix.
• Limit of Quantitation (LOQ) is the smallest
amount of the analyte that can be quantified with
a certain degree of reliability.

10. • Lowest level of Method Validation (LLMV) is the
lowest fortification concentration level at which
the method is validated for a particular matrix.
Definitions

11. Reference Substance
• Section 28 in an IR-4 protocol describes how to
obtain reference substance and the contact
information.

12. Reference Methods
Section 29.1 in an IR-4 protocol includes a reference
method (registrant reference method) that mainly
should be followed.

13. Working Method

14. Working Method Example

15. Change to Method
• Due to the nature of sample matrices and
availability of analytical instrumentation, a
working method can be developed with
“minor modifications”.
• The working method is needed to verify the
validity of the method for different matrices.
• The SD approves the “minor modifications”.

16. Minor vs Major Changes
“Minor Modifications” are generally:
Changes in sample size, which should not lead to
sample integrity and homogeneity concerns
Changes in extract volumes; if it is a change in sample
size, the change of the rest procedure should be
changed accordingly and is typically proportional to
those the original method
Modification of cleanup steps
Removal of cleanup steps
Optimization of instrument analysis parameters

17. Minor vs Major Changes
“Major Modifications” are:
Changes in extraction method or extraction
solvent
Changes in chemistry at major steps
(extraction, derivatization, hydrolysis,
detection nature, etc.) .

18. Q: Minor or Major ?
Change in extraction method?
Homogenizing QuEChERS
Change in extraction solvent?
Acetonitrile  Acetone

19. Q: Minor or Major ?
Change in Instrument?
GC/MS  HPLC/MS
Change in Instrument?
HPLC / UV LC/MS/MS

20. Weighing
• Balance annual calibration (certificate)
• Check balance using standard weights
• Make sure all weights are bracketed by standard weights

21. Weighing
• Sample jars are well labeled
• Double sample ID before weighing
• Weigh frozen samples (do not let it thaw)

22. Balance Log
Minimum 1 control
Minimum 1 QC
Field samples
Calibrate pipet
(if pipet is used)
Check balance
before weighing
weigh vessle to
bracket weighings

23. Fortification
• Pipet/syringe calibration
• Fortify samples

24. Method Validation (MV)
• Three (3) recovery spike concentrations in
triplicates are required as per protocol.
• Lowest level of method validation (LLMV) and
two higher concentrations. Bracket anticipated
range of residues.

25. Inform SD with MV Data
• The SD is informed of the
recovery results and the
working method.
• Recoveries of 70 - 120%
are accepted.
• If outside this range, SD
approval or additional MV
recovery spikes are
required.

26. Question: GLP Starts in Lab
• When GLP starts in an analytical lab?
 SD signs the protocol
 Analytical reference standard is received and the first
application is performed in the field
 First field sample is received by lab
 MV is completed and SD approves to start sample
analysis

27. Sample Analysis

28. Analytical Set
An analytical set
contains:
- Minimum 1 control
- Minimum 1 QC
(concurrent recovery spike)
- Field treated samples.

29. Double Injection
• Double injection (also called dual injection) is
required for analyzing weathered samples,
including control, concurrent spike and field treated
samples in the same analytical set. For example,
each sample vial is injected for analysis by two
separate injections (as one after the other) to
ensure the quality of analytical data.
• Double injection is not required for analytical
standards, solvent blank, samples of MV set, or
samples of SS sets.

30. Worklist (Sequence)
• Calibration standards1
• Solvent blank2
• Control sample3
• Concurrent spike sample(s) 4
• Field treated samples5
• Solvent blank2
• Calibration standards1
1 Generally 5-6 calibration standards (minimum 3), starting and ending the worklist
2 Minimum one blank after standards; may add more to ensure minimized carryover
3 Double injection is require for control analyzed with treated samples Order may change
4 Double injection is required for QC sample (minimum 1 QC at LLMV)
5 Double injection is required for treated samples

31. Worklist (Sequence)-Example

32. Calculation Sheet (Example1)

33. Calculation Sheet (Example2)

34. Sample Analysis
• During the course of residue sample
analysis, adequate concurrent recovery
spikes bracketing the actual residues have
to be analyzed.
• If recoveries outside of 70 - 120%, set will
first be re-injected and if still not
satisfactory another set will be extracted
and reanalyzed in agreement with the SD.

35. Lab Records
• Controls with interference or
contamination as well as field
samples with unusual
residues must be reported to
the SD promptly.
• During MV as well as during
field sample analysis
instrument parameters and
conditions should stay
constant and are recorded in
an instrument log book.

36. Data Acceptance Criteria
• Spike recovery is between 70 - 120% 1
• Double injection on weathered samples 3
• Meet linearity requirements (R2 ≥ 0.98 or 0.99) 2
• Five (5) calibration levels or more 2
• Difference between double injection ≤ 15% 2,3
• Days between extraction and analysis ≤ 14 days1
• Residues in treated bracketed by spike levels1,3
• Residue in control ≤ LOD 3
1 Required by protocol
2 Required by Lab SOPs
3 IR-4 Laboratory Guidelines

37. Storage Stability

38. Storage Stability
• One time analyses are
carried out to show that
samples are stable under
the conditions stored.
• Storage stability analysis
is performed after the
longest interval between
harvest and extraction of
a field sample.

39. Storage Stability

40. Calculation of LOD/LOQ
• Before completion of
the analytical phase
of the study a
minimum of six
recovery spikes at
the LLMV are
required in order to
calculate the LOD
and LOQ.

41. LOQ Calculations

42. Analytes of Interest?
• Residue Expression will determine which
compounds (Parent, metabolites) will need to
be analyzed for risk assessment and
enforcement
• Determined by Regulatory authorities after
extensive review of metabolism and toxicology
data

44. True or False
A copys of an original raw data page directly
from a Canon photocopier is an exact copy of
the raw data.
A computer printout derived from data
transferred to computer media from laboratory
data sheets is considered to be raw data.

45. True or False
GLP studies may be done in facilities that
utilize other quality standards (GMP, GCP, ISO
etc.).
EPA inspector can take photographs or
photocopies of objectionable practices and
conditions.

46. Q & A
Raw data must be co-signed by a second
individual.
Raw data must be directly entered into bound
notebooks

47. Q & A
QAU must keep a copy of lab SOPs.
Study director authorizes changes in the SOPs.

48. Q & A
What are the GLP requirements for labeling of
solvents purchased directly from
manufacturers?
(select all correct answers)
(A) Expiration date
(B) Identity
(C) User’s initial and date
(D) Safety information
(E) Storage condition.

49. Q & A
Which role can a study director be ?
(Select all correct answers.)
(A) Archivist
(B) Field Research Director
(C) Lab analyst
(D) Company CEO
(E) QA

50. jiangwa@msu.edu

51. Form QA5 Revised effective 1/31/10 SOP 8.7 Rev. 5 App. A
A) Study Title:____________________________________________________________
Lab I.D. Number: ______________________________________________________
Inspection Date: ________________________________________________________
B) Laboratory Research Director:
Name:_________________________________________________________________
Address:_______________________________________________________________
Phone: ( )__________________________________________________________
E-mail:________________________________________________________________
C) Regional / ARS Laboratory Research Coordinator:
Name:_________________________________________________________________
Address:_______________________________________________________________
Phone: ( )__________________________________________________________
E-mail:________________________________________________________________
D) Quality Assurance Inspector:
Name:_________________________________________________________________
Address:_______________________________________________________________
Phone: ( )__________________________________________________________
E-mail:________________________________________________________________
E) Study Director:________________________________________________________
E-mail:________________________________________________________________
F) Test Site Location:______________________________________________________
______________________________________________________________________
G) Please fill out the following checklist.
Provide a narrative on any items marked No and provide suggestions and recommended
actions to be taken as appropriate for all QA findings. Use additional forms if needed.
Study personnel must respond to QA Findings.
Please Note:
Any problems which are likely to affect the study’s integrity found during the course of the review must
be brought to the attention of the Study Director immediately.
IR-4 PROJECT
Analytical Raw Data Audit
□ Circulate to TFM/SD simultaneously
Page 1 of _____

52. Form QA5 Revised effective 1/31/10 SOP 8.7 Rev. 5 App. A
Page 2 of ____
Analytical Raw Data Audit
ID #________________
A. General YES NO N/A
* 1. Approved protocol and method included in raw data package.
* 2. Changes were authorized by the Study Director per protocol.
* 3. Method used for analysis was validated per protocol prior to
use in analyzing study samples.
* 4. All modifications to the referenced method were documented,
validated, signed and dated by the LRD (working method).
* 5. All SOP deviations listed in the raw data.
* 6. All SOP deviations authorized by Study Director.
* 7. Appropriate personnel signatures included in raw data.
* 8. All data corrections properly explained, initialed and dated.
* 9. All pages properly identified.
*10. Procedures used in generating raw data were described in the
SOPs, protocol and / or study raw data.
B. Sample Storage and Preparation YES NO N/A
*11. Samples traceable through chain of custody documentation.
a. Receipt.
b. Storage.
c. Distribution.
*12. Sample preparation according to SOP.
*13. Sample preparation adequately recorded.
*14. Sample preparation followed validated method.
*15. Sample storage location(s) documented.
*16. Sample storage temperatures documented.
*17. Date and times samples taken in and out of the freezer logged
and within SOP requirements.
*18. Storage duration and conditions of storage of samples &
stability samples are the same.
* Minimal GLP requirements associated with Series 860.
N/A = Not applicable
Comments

53. Form QA5 Revised effective 1/31/10 SOP 8.7 Rev. 5 App. A
Page 3 of ____
Analytical Raw Data Audit
ID #_______________
C. Analytical Reference Standards and Fortification Solutions YES NO N/A
*19. Check all analytical standard used, source, batch numbers and
expiration dates for acceptability.
*20. Certified copy of certificate of analysis for standard(s) is in the
study file.
*21. Standards solution was used prior to their expiration dates.
*22. Accountability of reference standards.
a. Records and receipts.
b. Use logs up-to-date (distribution and disposal).
c. Storage logs.
d. Storage location(s).
e. Storage conditions.
*23. Laboratory raw data documents the standards used (proper
identification maintained).
*24. Retention sample of the standard is in an IR-4 Laboratory
chemical archive or archive facility is documented.
*25. Logbook(s) for balance (s) contain(s) calibration
documentation.
*26. Standard solutions documentation adequate.
a. Stock.
b. Analytical standards
c. Fortification solutions.
*Minimal GLP requirements associated with Series 860.
N/A = Not applicable
Comments

54. Form QA5 Revised effective 1/31/10 SOP 8.7 Rev. 5 App. A
Page 4 of ____
Analytical Raw Data Audit
ID #________________
D. Data Inspection YES NO N/A
*27. Raw data properly recorded.
a. Promptly and legibly in ink.
b. Dated on day of entry and signed or initialed.
c. Changes to entries did not obscure the original.
d. Corrections were explained, date, and signed or initialed.
*28. Computer generated data.
a. Program has been validated.
b. Input personnel identified.
c. Printout signed.
d. Printout dated.
*29. Numerical results reported were consistent for significant
figures, rounding-off numbers, etc.
*30. Units of concentration were clearly identified.
*31. Instrument parameters were documented for each set of runs:
a. Instrument conditions / date.
b. Study number.
c. Lab sample / standard concentration.
d. Analyst(s) / operator(s) initials.
e. Injection volume.
*32. Injection sequence. (all chromatograms retained in continuous
sets per run).
*33. Samples fall within standard curves range.
*34. Chromatograms and standard curves audited.
*35. Integrator chromatograms and / or computer generated
chromatograms compared to data report.
*36. Analytical instrument logbooks showed proper documentation
and operation.
*37. Analytical sets, including standards and fortifications according
to SOPs and protocol.
*38. Limits of quantization and detection (LOQ and LOD) were
clearly defined.
*39. Calculations were accurate.
*40. Recoveries outside of 70-120 range documented and authorized
by LRD and study director.
* Minimal GLP requirements associated with Series 860.
N/A = Not applicable
Comments

55. Form QA5 Revised effective 1/31/10 SOP 8.7 Rev. 5 App. A
Page _______ of ______
Study Title:
Study No.:___________________________ Study Dir.:
Findings/Actions
QA Findings Response/Actions Taken*
______________________________ ______________________________
______________________________ ______________________________
______________________________ ______________________________
______________________________ ______________________________
______________________________ ______________________________
______________________________ ______________________________
______________________________ ______________________________
______________________________ ______________________________
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______________________________ ______________________________
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______________________________ ______________________________
______________________________ ______________________________
______________________________ ______________________________
____________________________________ ____________________________________
Quality Assurance Date Responder** Date
* - Place responses/ explanation of corrective action in space provided to the right side of the findings or
use a separate sheet of paper.
** Responder(s) are to assure they have identified themselves either by signing the bottom of this page
or by initialing and dating the written response(s).

56. Form QA5 Revised effective 1/31/10 SOP 8.7 Rev. 5 App. A
Page ______ of ______
Study Title:__________________________________________________________________
Study No.: _________________________________ Study Director:
Findings, Responses and Actions Taken*:
____________________________________ ____________________________________
Quality Assurance Date Responder** Date
* - Place responses/ explanation of corrective action in space provided between findings or use a
separate sheet of paper.
** Responder(s) are to assure they have identified themselves either by signing the bottom of this page
or by initialing and dating the written response(s).