Good Laboratory Practice

ALS 5204

PRESENTED BY

JEAN COBB

SENIOR PROJECT ASSOCIATE

PESTICIDE RESIDUE LAB

BIOCHEMISTRYFor Field and Laboratory Research

What are GLPs?

Good Laboratory Practice standards (GLPs) are federal regulations mandated in the United States by:

• Food and Drug Administration (FDA) in 21 CFR Part 58

• Environmental Protection Agency (EPA) both for FIFRA in 40 CFR Part 160 and for TSCA in 40 CFR Part 792

Key Dates1970 1976 FDA GLP proposed

1978 FDA GLP regulations published1980 1982 OECD GLP published (30 countries incl.

U.S.), UK GLP published, Japanese GLP published

1983 EPA GLP regulations published1986 FDA GLP revised, European Union GLP Directives1989 EPA GLP revised

1990 1998 OECD GLPs revised, Canadian PMRA adopted GLPs

2000 2002 OECD consensus document 2007 EPA may consolidate two parts of EPA GLPs

Where are the EPA GLP Regulations Published?

National Archives and Record Administration

Code of Federal Regulations (CFR)

Title 40 Protection of Environment (laws and regulations of U.S. EPA)

Part 160: Good Laboratory Practice Standards, Federal Insecticide, Fungicide and Rodenticide Act (FIFRA), Subparts A-J

Part 792: Good Laboratory Practice Standards, Toxic Substances Control Act (TSCA), Subparts A-J

Note: FIFRA regulates pesticides and TSCA regulates toxic substances

Why Were the GLPs Mandated?

• In the 1970s, numerous cases of laboratory malpractice surfaced in toxicity testing of food and drugs in the U.S.

• FDA inspectors found inaccurate, sloppy, and fraudulent research studies

• Became a public safety issue

One of the first names to attract regulatory

and media attention was IBT…

IBT: Industrial Bio-Test Laboratories

Illustration by Dan Sherbo©

Problems at Test Facilities

Many of IBT’s (Industrial Bio-Test Laboratories) test animals were housed in a water-logged room known as the Swamp.

“ During the course of a two-year feeding study, involving more than 200 animals, the mortality rate in the Swamp reached 80 percent. Dead rats and mice…decomposed so rapidly in the Swamp that their bodies oozed through wire cage bottoms and lay in purple puddles on the dropping trays.”

- The Amicus Journal, Spring 1983, p.14

Problems with Sponsors

G. D. Searle Co. (Pharmaceutical Manufacturer)

“…’Because of the perfunctory nature of the observations, tissue masses come and go and animals die more than once.’ In fact some rats listed as dead later were recorded as alive, then dead, then resurrected once or even twice more.”

– The Impact of Publicity on Corporate Offenders, Ch. 12, p. 138

The Regulatory ProcessBefore GLPs – Few “checks and balances”

TestFacility

Sponsor Approve?

Submits Data Regulatory

Agency (EPA, FDA)

Yes! Permission to

Market Regulated Product

No!Permission to

Market Regulated Product Denied

Industrial Bio-Test Laboratories (IBT), Craven Labs, Intertek Testing Services, & others

Searle Co. ChemAgro Monsanto & others

Test Facility: Field and laboratory (contract) research facilities

Sponsor: Pharmaceutical and pesticide manufacturers

TestFacility

Sponsor Approve?

Submits Data Regulatory

Agency (EPA, FDA)

Yes! Permission to

Market Regulated Product

No!Permission to

Market Regulated Product Denied

Inaccurate, sloppy, and fraudulent science???

Poor Management Practices ????

Would you want the FDA to issue a permit to manufacture of a drug based on sloppy research? Or for the EPA to register a pesticide based on fraudulent data? Probably not!

The Regulatory Process Before GLPs – Few “checks and balances”

What Role Do GLPs Play?

Good Laboratory Practice standards help to assure the quality and integrity of studies submitted in support of FDA and EPA regulated products

TestFacility

Sponsor Approve?

Submits Data Regulatory

Agency (EPA, FDA)

Yes! Permission to

Market Regulated Product

No!Permission to

Market Regulated Product Denied

Internal audits and inspections (conducted by Quality Assurance Unit)

The Regulatory ProcessAfter GLPs

External audits and inspections of sponsor and test facility (conducted by regulatory agency)

GLP Regulations

Greater scrutiny of field and laboratory studies of regulated products

Auditor/ Inspector

Who must comply with GLPs?

FDA GLP Compliance

Required:

Pre-clinical safety studies for the development of food additives and drugs (before a marketing permit is issued)

Regulated products: Pharmaceuticals, cosmetics, food & color additives, human medical devices

Note: “pre-clinical” refers to safety testing in animals, plants, or microorganisms (not humans)

EPA GLP Compliance

Required:

Field and laboratory studies of pesticides (before a marketing permit or application for research is issued)

Regulated products: Pesticides and other substances that could spread during agricultural practice

GLP Regulations: Rules and Tools

Chemical and sample inventory, track expiration dates, labeling

TEST, CONTROL, AND REFERENCE SUBSTANCES

Timely reporting, storage of raw data and reports

RECORDS AND REPORTS

Standard operating proceduresFACILITY OPERATION

Calibration, logbooks of use, repair, and maintenance; check freezers

EQUIPMENT

Maintain adequate space/separation of chemicals from office areas

FACILITIES

Training records, CVs, GLP trainingORGANIZATION AND PERSONNEL

Documentation (Tools)GLP Regulations (Rules)

Documentation

If you didn’t write it down, it NEVER happened. (As far as an auditor is concerned…)

Auditor/ Inspector

Equipment

40 CFR Part 160 (EPA GLP regulations)

“Section 160.63 Maintenance and Calibration of Equipment. (a) “Equipment used in the generation, measurement, or assessment of data shall be adequately tested, calibrated, and/or standardized. (c) Written records shall be maintained of all inspection, maintenance, testing, calibrating, and/or standardizing operations….”

Do you know the difference between standardized, calibrated, and verified?

Equipment

• Verification (Testing): external check of equipment accuracy (check balance accuracy against weights at laboratory- no adjustment)

• Calibration: equipment is adjusted based on comparison to certified or known reference materials (balance adjusted after comparison to certified weights by trained professional)

• Standardization: comparison with similar equipment (use two thermometers of similar design to compare readings)

Equipment

What about validation?

• Electronic data management systems (computers, instrumentation) must be “validated” to confirm that they will perform their intended functions

• For example, a computer spreadsheet used for calculations during a GLP study could be validated as follows: enter identical raw data as previously and compare the calculations using the same software/options- they should be identical!

Use Logbooks

• Record all pertinent operating parameters

• Take the time to record routine service and how problems were resolved (it may help if/when the same problem arises!)

AGILENT GAS CHROMATOGRAPH SERIES: 6890n VT#0000322580

N/ANew glass insert & gold seal 9/17/07 S.J.

He-carrier, 1.0 mL/min, H2/Air det. gases

300°C225°C60-275°CRTX-OPP (30 m x 0.25 mm x 0.25 µm)

NPDN/AStandards only

Sally Jones (lab tech)

9/17/07

Power outage caused GC shutdown

N/AHe-carrier, 2.3 mL/min, N2 makeup

350°C250°C90-275°CRTX-5 (30 m x 0.25 mm x 0.25 µm)

µECD #8128/ µECD #5663 (dual column)

Plant Tissue

VCE07-021

Sally Jones (lab tech)

9/14/07

N/AReplaced He gas - VAC052 9/13/07 J.E.S.

He-carrier, 2.3 mL/min, N2 makeup

350°C250°C60-275°CRTX-5 (30 m x 0.25 mm x 0.25 µm)

µECD #8128/ µECD #5663 (dual column)

WaterVCE07-020

John Smith (lab tech)

9/13/07

PROBLEM (#)

ROUTINE SERVICE (#)

GAS mL/minDET TEMP °c

INLET TEMP °c

OVEN TEMP °c

COLUMN TYPE/ SIZE

DETECTOR TYPE

SAMPLE MATRIX

PROJECTNAMEDATE

• Was the equipment functioning properly? Who performed the work, what was the date, and what specific parameters did they use?

• What was there a problem? How was the problem fixed?

Equipment

Reagents and Solutions

40 CFR Part 160 (EPA GLP regulations)

“Section 160.83 Reagents and Solutions. “All reagents and solutions in the laboratory areas shall be labeled to indicate identity, titer or concentration, storage requirements, and expiration date. Deteriorated or outdated reagents and solutions shall not be used.”

How do you know what expiration date to use?

Reagents and Solutions

General expiration guidelines:• Dry reagents: 5 years (unless reevaluated)

• Solvents: 1 year from date opened or 2 years from date received

• Solutions: Use the earliest date of components in the solution or no longer than 6 months

Question: Which takes precedence: the manufacturer’s expiration date or expiration date as stated in the facility SOPs?

Answer: Whichever is earliest!

Use Labels

• Clearly label chemicals and solution containers

• Make sure that the expiration date and storage requirements are included on the label

• If a solution, what date was it prepared?

CHEMICAL LABEL

CHEMICAL___ ____________ ________

CHEM ID#______________________________ _

DATE RECEIVED:___________________________

DATE OPENED: ____________________________

EXPIRATION DATE__ _ _______STORAGE______

SOLUTION LABEL

CONCENTRATION_____________________

SOLUTION____________________________

FROM CHEM ID#_____________________

DATE PREPARED_____________________

EXPIRATION DATE______ STORAGE______

Use Forms• Assign a unique number

to each chemical received by the facility

• Record the purity, date received, storage conditions, and source

• Use the form to “jog your memory” about the information you need to collect

PESTICIDE RESIDUE LABORATORY

2007 CHEMICAL INVENTORY FORM

LAB CHEMICAL ID (s): __C070014______________

CHEMICAL NAME: ethyl acetate

CAS NO.: 141-78-6

SOURCE: Fisher Scientific

LOT NO.: 067720

PURITY/GRADE: Pesticide Grade

QUANTITY RECEIVED: 4 L

EXPIRATION DATE: 9/7/12

STORAGE CONDITIONS: ambient, in solvent cabinet

DATE RECEIVED: 9/7/07

SIGNATURE: Jean M. Cobb

COMMENTS: N/A

DATE INSPECTED/METHOD: N/A

NEW EXP DATE: N/A

Test, Control, & Reference Substances

• Are the test, control, and reference substances of known quality and purity?

• Are these substances used within expiration dates?

• Are they adequately stored?

Raw Data

Question:What happens if you make a mistake?

Answer:

Do not obscure original data!! Instead, draw a single strikeout, then add reason code, initials, and date of change. (Preserve the original data).

Documentation

• Record data using permanent ink (never pencil)

• Date and sign every entry (who is responsible?)

• Keep records in “real time” (no catching up later)

• No rewrites, need original entries (reduces transcription errors)

Documentation is important in all sorts of situations!!

Raw Data

40 CFR Part 160 (EPA GLP regulations)

“Section 160.3 Definitions. “’Raw data’ means any laboratory worksheets, records, memoranda, notes, or exact copies thereof, that are the result of original observations and activities of a study and are necessary for the reconstruction and evaluation of the report of that study.”

If you scribble some notes on a scrap of paper, are those notes considered raw data? Answer: Yes!

Raw Data

Can you list examples of raw data?

• Logbooks (to record temperatures or equipment use, repair, and maintenance)

• Field or laboratory notebooks• Forms (for field or laboratory observations, chain-of-

custody, sample or chemical receipt)• Training reports• Computer printouts• Recorded data from automated instruments

How can anyone remember all these details?

Answer: You don’t have to… that’s one of the functions of the Standard Operating Procedures (SOPs).

Write Standard Operating Procedures (SOPs)

SOPs are written, approved procedures that describe routine activities that are specific for daily operations at each facility.

SOPs should allow appropriately qualified personnel to perform a procedure once trained.

Standard Operating Procedures

40 CFR Part 160 (EPA GLP regulations)

“Section 160.81 Standard operating procedures. (a) A testing facility shall have standard operating procedures in writing setting forth study methods that management is satisfied are adequate to insure the quality and integrity of the data generated in the course of a study.”

Personnel perform the same tasks using the same procedures.

SOP Writing Guidelines

• SOPs should accurately reflect how routine tasks are performed (written by each facility based on their specific field and/or laboratory operations)

• Make sure they are useful in daily operations

• They should be scientifically sound

• Remember to update as necessary, rewrites are part of the process

Writing Guidelines

Cautions:

• Avoid restrictive language such as “vortex for exactly 1 minute” but include clear instructions such as “vortex until homogenized” if that satisfies the purpose

• Don’t add unnecessary steps such as “consult the manual” unless personnel are required to follow this step

Writing Guidelines

The primary goal is to avoid confusion…

Did your supervisor say to make a 1 in 4 dilution… or was it a 1 to 4 dilution?

If it is written in an SOP, the instructions can be verified.

What if you don’t have to comply with GLP

regulations?

GLP Compliance

Not Required:

• Basic research (most of the research conducted at universities like Virginia Tech)

• Studies to develop new analytical methods

• Chemical tests used to derive the specifications of a marketed food product

• Instead, “emphasize GLP principles” during research

GLP Principles

GLP principles are a good idea even if you are not required to comply with the regulations.

• Say What You Do (with written standard operating procedures)

• Do What You Say (follow the procedures)

• Be Able to Prove It (with good record keeping)

Is GLP Experience Important to Employers?

Yes! In some cases…

GLP Experience

An employer may find it useful if you have:

• Practical experience with working on a study conducted in compliance with GLP regulations

• Experience working at a facility that emphasizes GLP principles

• Familiarity writing and using standard operating procedures at a GLP facility

• Everyone makes mistakes

• Mistakes cost $$$

• Increasing the quality of an operation is good for business and science

Why Do We Need GLPs?

What GLPs SOPs Can’t Do

• Guarantee “good science”

• Guarantee good documentation

• Replace common sense

• Prevent all mistakes

What GLPs Can Do

• Fulfil the legal requirements of some FDA and EPA research studies

• GLP principles can be followed to increase the quality of basic research

Websites

• Code of Federal Regulations (CFR)- all

• EPA GLP Regulations

• FDA GLP Regulations

• USDA- Pesticide Data Program (PDP) Standard Operating Procedures