This standard operating procedure outlines the process for estimating alanine aminotransferase (ALT) levels in serum or plasma samples. It describes the scope, purpose, responsibilities, requirements including specimens, materials and equipment, procedure, reporting including reference ranges and clinical interpretations, quality assurance, and reference documents. The procedure is to be followed by lab technicians, assistants, attendants and overseen by quality assurance and lab supervisors to ensure consistent and accurate ALT testing that adheres to standard protocols.

1. Standard Operating Procedures
(SOPs)
Lecture 9
Dr. Attya Bhatti
HoD Health care Biotechnology
ASAB-NUST

2. Standard Operating Procedures (SOPs)
• ‘Detailed, written instructions to achieve uniformity of
the performance of a specific function’.
• Focus is always set on repeated application of unchanged
processes and procedures.

3. Purpose of SOP
• Serve as framework for organizational policy – provide
direction and structure
• Written documentation of best practice
• Tells what, how, when, why, and who
• Provide foundation for:
▫ job descriptions,
▫ employee training
▫ corrective action and discipline, and
▫ performance review.

4. • Help maximum safety and operational efficiency
for organisations:
▫ Pharmaceutical organizations
▫ Government organizations
▫ Emergency response operations
▫ Clinical research organizations
▫ Power producing organizations

5. Who is responsible?
• Study director is mainly responsible for SOPs.
• The Quality Assurance Unit are individuals who
are responsible for monitoring whether the
study report and tests are meeting the SOP.

6. How are SOP's developed?
• The Four P's Model
▫ Philosophy
▫ Policies
▫ Procedures
▫ Practices

7. Elements of an SOP
• Rationale for SOP
• Detailed description of procedure – based on best
practice/standards
• Monitoring actions
• Accountability
• Corrective Actions
• Date of last review or revision date

8. Writing Styles
•SOPs should be written in a concise, step-by-step, easy-to-read format.
•The information presented should be unambiguous and not overly
complicated.
•The active voice and present verb tense should be used. The term "you" should
not be used, but implied.
•The document should not be wordy, redundant, or overly lengthy.
•Keep it simple and short.
•Information should be conveyed clearly and explicitly to remove any doubt as
to what is required.
•Also, use a flow chart to illustrate the process being described.
•In addition, follow the style guide used by your organization, e.g., font size and
margins

9. SOP GENERAL FORMAT
• SOPs should be organized to ensure ease and efficiency in use
and to be specific to the organization which develops it.
• There is no one “correct” format; and internal formatting will
vary with each organization and with the type of SOP being
written.
• Where possible break the information into a series of logical
steps to avoid a long list.
• The level of detail provided in the SOP may differ based on, e.g.,
whether the process is critical, the frequency of that procedure
being followed, the number of people who will use the SOP, and
where training is not routinely available.

10. Title Page
• The first page or cover page of each SOP should contain the
following information:
▫ A title that clearly identifies the activity or procedure,
▫ An SOP identification (ID) number,
▫ Date of issue and/or revision,
▫ The name of the applicable agency, division, and/or branch to
which this SOP applies,
▫ The signatures and signature dates of those individuals who
prepared and approved the SOP.
 Electronic signatures are acceptable for SOPs maintained
on a computerized database.

11. Table of Contents
A Table of Contents may be needed for quick
reference, especially if the SOP is long.

12. Text
• Well-written SOPs should first briefly describe
the purpose of the work or process, including
any regulatory information or standards that are
appropriate to the SOP process, and the scope to
indicate what is covered.
• Define any specialized or unusual terms either in
a separate definition section or in the
appropriate discussion section.

13. Text
• Denote what sequential procedures should be
followed, divided into significant sections; e.g.,
possible interferences, equipment needed,
personnel qualifications, and safety
considerations (preferably listed in bold to
capture the attention of the user).
• Finally, describe next all appropriate QA and
quality control (QC) activities for that procedure,
and list any cited or significant references.

14. SOPs Implementation
• Have available for workers to review
• Use to train new employees
• Use them for corrective action/refresher training for all
employees
• Incorporate into written job descriptions and
performance reviews so there is alignment
• Review and update as needed (i.e new equipment item or
employee job changes); at minimum review annually

15. Apply ALCOA to achieve data quality.
1. Attributable: is it obvious who wrote it?
2. Legible: can it be readable?
3. Contemporaneous: is the information
current and in the correct time frame?
4. Original: is it a copy; has it been altered?
5. Accurate: are conflicting data recorded
elsewhere?
http://www.ofnisystems.com/growing-need-
for-good-data-and-record-management/
ALCOA

16. The SOPs can be categorized
SOPs
Instrument
SOPs
Protocol
SOPs
Solution
SOPs

17. Instrument SOPs (S-SOP)
• Procedures describing how to use our
instruments
• e.g. Fume Hoods, Thermo cyclers, Gel
Documentation System

18. Protocol SOPs (P-SOP)
• Procedures describing methods
▫ e.g. DNA isolation, RNA isolation, Agarose gel
electrophoresis, Cell proliferation assay, ELISA
etc.

19. Solution SOPs (S-SOP)
• Procedures describing preparation of solutions
▫ e.g. 1M TRIS, DNA extraction solution, MTT
reagent, Bradford reagent, Cell lysis buffer,
Loading dye etc.

20. Factors Affecting SOP Development
• Factors that need to be considered for the SOP's
to work in the real world environment.
▫ Organizational Factors
▫ Operational Factors

21. The pros of SOPs
• Consistency
▫ Must be consistent;
 consistency in the performing of an operation,
whether it is in manufacturing or department
administration. The more consistent the process is
from person to person, the less chance there will be
quality problems.

22. The pros of SOPs
• Reduction of Defects
▫ Reduction of defects is another reason for operating
procedures.
▫ A written procedure is like a set of instructions for
performing a task. As long as each person performs
the task exactly as the process is written, there should
not be any defects.

23. The pros of SOPs
• Communication
▫ Another reason for standard operating procedures is a
method of communication.
▫ As improvements are made to processes, the operating
procedures are updated and each update requires new
training. This provides a method to communicate the
process changes to all employees.

24. SOP # :
PHLD/ CP/003/01
Revision # : 01 Replaces: 00
Written by:
Dr. ABC
Edited by:
Dr. XYZ
Authorized by:
Dr. 123
Effective Date:
November 2010
1. Scope:
This procedure is applicable to Chemical Pathology Lab of PHLD when
ALT estimation is performed.
2. Purpose:
To streamline the quantitative in vitro determination of serum ALT level
through appropriate verification & adequate documentation.
To ensure and monitor the operational phases of ALT estimation in patients blood
using standard protocol.

25. 3. Responsibility:
1. Lab Technician with the help of Lab Assistant & Lab Attendant is
responsible to prepare sample for testing, to run both normal and
abnormal high controls and ensure control results fulfill the QA criteria
to perform the assay as per protocol
2. Quality Assurance supervisor is responsible to control and manage lab
activities in order to ensure efficient processing of requests and timely
delivery of service and to scrutinize the final reports.
3. The Officer Incharge/Pathologist is responsible for overall monitoring
and providing support and guidance in problem solving.

26. 4. Requirements
4.1 Specimen
 Serum, plasma. (fresh or hemolytic)
4.2 Materials
Kit from DiaSys diagnostic systems
Roche Multical
Controls (Roche PPU, PNU)
Sample cups
4.3 Equipment:
Vitalab Selectra E Auto Analyzer
Micro lab 300
Pipettes of various ranges
Centrifuge Machine

27. 5. Procedure:
5.1 Collection, Transportation and Accessioning of Chemical Pathology samples.
(see SOP no…..)
5.2 Principle:
L-Alanine + 2-Oxaloacetate ALAT L-Glutamate + Pyruvate
Pyruvate + NADH + H LDH D-Lactate + NAD
5.3 Method:
Referred to Kit Literature.
Referred to EOP Selectra E.
5.4 Linearity:
The test has been developed to determine ALT activities which correspond to a Maximal ∆A/min of 0.16 at 340
and 334 nm or 0.08 at 365nm. When values exceed this range samples should be diluted 1+9 NaCl solution( 9g/L) and
result multiplied by 10.
5.5 Interfering Factors and sources of errors:
1. Many drugs can cause elevated level.
2.Salicylates may cause increase or decrease ALT levels.
5.6 Stability: In serum/plasma
3 day at 20-25˚C
7 days at 4-8˚C
7 days at -20˚C

28. 6. Reporting:
6.1 Reference Range:
In serum/plasma:
With pyridoxal-5-phosphate activation
Women[3] < 34 U/L
Men[3] -< 45 U/L
Children[1] 1-30 days < 25U/L
2-12 monhs < 35 U/L
1-3 years < 30 U/L
4-6 years < 25 U/L
7-9 years < 25 U/L
10-18 years < 30 U/L
Without pyridoxal-5-phosphate activation
Women < 31 U/L
Men < 41 U/L
6.2 Clinical interpretations:
i) Increase ALT levels is significant in
a. Hepatocellular disease
b. Active cirrhosis
c. Metastatic liver tumour
d. Obstructive jaundice
e. Viral, infectious or toxic hepatitis

29. 7. Quality Assurance:
(Ref SOP # ******** on QA Protocols and tools)
8. Reference documents:
Fischbach, F.T.1996.Immunodiagnostic studies. In: A manual of
Laboratory and diagnostic tests.5th ed. JB Lippincott company,
Philadelphia, PA, USA.pp554 Kit literature