PFT869 - Method Development

Outline info
Last revision date 2018-07-20 11:49:59.964
Last review date 2018-07-20 11:50:28.981

Subject Title
Method Development

Subject Description
This subject is designed for students who already have an understanding of fundamental HPLC concepts and would like to focus on method development techniques. Topics include review of HPLC basics, establishment of the initial HPLC conditions, systematic HPLC method development and HPLC troubleshooting. Students have opportunities to learn method development techniques such as the triangle MP optimization approach and establishment of isocratic conditions through scouting gradient elution. The subject prepares students with the method development skills needed to work in the R and D environment independently. Laboratory practice reinforces theory. (Lectures and labs)

Credit Status
One credit toward the Advanced Pharmaceutical Instrumentation (HPLC) Recognition of Achievement

Learning Outcomes
Upon successful completion of this subject the student will be able to:

1. Demonstrate proficiency in the fundamental concepts of HPLC method development.

2. Optimize HPLC conditions at different stages of method development, including sample handling, selection of column and mobile phase, and detection techniques.

3. Apply a logical and systematic approach to HPLC method development.

4. Examine modern tools available to the method development chemist (e.g. use of column concentrator).

5. Assess the roles of various HPLC method parameters such as solvents ratio in mobile phase, pH, temperature, column selection and their impact on peak retention.

6. Demonstrate an understanding of the concepts of pKa, buffer pH, organic solvents and their importance in developing a method for ionic or ionizable samples.

7. Demonstrate an understanding of the concept of ultraviolet absorption and its optimum wavelength.

8. Explain the concept of reverse phase, normal phase, ion pair and ion exchange chromatography.

9. Apply HPLC theory and concepts effectively to troubleshoot HPLC separation problems.

Cheating and Plagiarism
Each student should be aware of the College's policy regarding Cheating and Plagiarism. Seneca's Academic Policy will be strictly enforced.

To support academic honesty at Seneca College, all work submitted by students may be reviewed for authenticity and originality, utilizing software tools and third party services. Please visit the Academic Honesty site on for further information regarding cheating and plagiarism policies and procedures.

All students and employees have the right to study and work in an environment that is free from discrimination and/or harassment. Language or activities that defeat this objective violate the College Policy on Discrimination/Harassment and shall not be tolerated. Information and assistance are available from the Student Conduct Office at

Accommodation for Students with Disabilities
The College will provide reasonable accommodation to students with disabilities in order to promote academic success. If you require accommodation, contact the Counselling and Disabilities Services Office at ext. 22900 to initiate the process for documenting, assessing and implementing your individual accommodation needs.

Work experience in HPLC or equivalent

Topic Outline
Introduction to chromatography, WHMIS overview

Method development  by HPLC and reverse phase chromatography as well as introduction to project # 1 (separation of neutral components)

Introduction to Empower software, creating methods in Empower for separation of acidic and neutral compounds

Project # 1 Method development to resolve 3 components mixture

Review of project # 1 for USP acceptance criteria and conclude with discussion based on the steps throughout the development

Solvent types, selection, purity requirements, buffer solutions, pH adjustments, and ion pair chromatography

Project # 2  Method development by ion pair chromatography using sulfonate salts

Continuation of project # 2 and discussion of the procedure

Project # 3  Method development by ion pair chromatography using Q series

Continuation of project # 3 and discussion of the procedure

Project # 4  Buffer preparation & pH adjustment to separate ionizable compounds

Continuation of project # 4 and fine tuning the development steps

Mode of Instruction
Lab and lecture  

Prescribed Texts

Reference Material

Required Supplies
Lab coat and goggles

Promotion Policy

Grading Policy
A+ 90%  to  100%
A 80%  to  89%
B+ 75%  to  79%
B 70%  to  74%
C+ 65%  to  69%
C 60%  to  64%
D+ 55%  to  59%
D 50%  to  54%
F 0%    to  49% (Not a Pass)
EXC Excellent
SAT Satisfactory
UNSAT Unsatisfactory

For further information, see a copy of the Academic Policy, available online ( or at Seneca's Registrar's Offices.

Modes of Evaluation

  • Assignments are due at the beginning of the class on which they are due.
  • A late penalty of 10% per day is assessed for late assignments, including those not handed in at the beginning of class when due.
  • Material will not be accepted after one week following the due date and/or when the marked material is returned to students, whichever comes first.
  • Assignments are to be prepared by computer.

Absenteeism and Exams
  • Students should be aware that absenteeism almost guarantees an inability to achieve satisfactory grades.
  • Students who are absent for an examination due to an emergency (e.g., motor vehicle accident, hospitalization or death in the family) may provide official documentation within five days of the missed exam and be provided a deferred exam at a later date.  Official documentation includes a death notice or an original doctor’s certificate identifying the date, length of time expected absence and the specific reason for the absence.  Examinations missed without official documentation and approval result in a grade of zero.
  • There are no deferred options for missed tests.  The value of missed tests, at the discretion of the Faculty, will be added to other evaluation components

English Proficiency
  • All written work should demonstrate the following characteristics for clarity and conciseness:
-writing is consistent with the rules of English grammar
-spelling and punctuation are correct
-sentences are structured correctly
-main ideas are supported with specific, relevant examples and reasons
-work flows logically through supporting statements/paragraphs
-work is arranged in correct format (e.g., as a report, essay)
-up to 10% of the final grade may be deducted on all work if the above English competencies are not met.

Format for Assignments
  • Students must use the standard, APA style for quoting sources.   Help is available at:

Laboratory Attendance

The laboratory component is essential and therefore it is strongly recommended  that you attend all labs.  Any missed labs must be supported with a legal document with three days of the lab.  Any student who fails to attend 2 scheduled laboratory classes for a 7 week subject and more than 3 laboratory classes for a 14 week subject will not pass the subject.    

Laboratory Safety
Students are required to review and understand the safety procedures and guidelines outlined on the first class and sign the sheet to this effect before beginning work in the laboratory.  Students must also wear a lab coat and safety glasses when conducting experiments.
A student who arrives without the proper safety equipment will not be permitted to participant in the lab but will be asked to leave the class.  The student will receive no grade for the lab missed.

Grading is based on the following marking scheme:
Laboratory 50%
Assignment 20%
Final Exam 30%

Other Evaluation Considerations
The student is expected to comply with the Safety Rules for working in the laboratory, sign the safety contract, and WILL NOT UNDER ANY CIRCUMSTANCES WEAR CONTACT LENSES in the laboratory.  The student will know where all safety equipment is located in the laboratory and will be familiar with WHMIS concepts and signage.


Approved by: Denis Gravelle