Course Title: Apply routine chromatographic techniques

Part B: Course Detail

Teaching Period: Term2 2009

Course Code: ONPS5211C

Course Title: Apply routine chromatographic techniques

School: 155T Vocational Health and Sciences

Campus: City Campus

Program: C5183 - Diploma of Laboratory Technology (Biotechnology)

Course Contact: Kirsten Balding

Course Contact Phone: +61 3 9925 4283

Course Contact Email:

Name and Contact Details of All Other Relevant Staff


Nominal Hours: 120

Regardless of the mode of delivery, represent a guide to the relative teaching time and student effort required to successfully achieve a particular competency/module. This may include not only scheduled classes or workplace visits but also the amount of effort required to undertake, evaluate and complete all assessment requirements, including any non-classroom activities.

Pre-requisites and Co-requisites

PMLTEST303B - Prepare working solutions OR PMLTEST402B - Prepare, standardise and use solutions
PMLTEST404A - Perform chemical tests and procedures.

Course Description

This unit of competency covers the ability to analyse samples using routine chromatographic techniques. The unit also includes establishing client needs for routine and non-routine samples, optimising enterprise procedures and instruments for specific samples, obtaining valid and reliable data and reporting test results. Personnel are required to recognise atypical test data/results and troubleshoot common analytical procedure and equipment problems.

National Codes, Titles, Elements and Performance Criteria

National Element Code & Title:

PMLTEST513A Apply routine chromatographic techniques


1. Establish client needs and schedule analysis
2. Prepare samples and standards
3. Set up and optimise instrument
4. Perform analysis
5. Process and analyse data
6. Maintain a safe work environment
7. Maintain laboratory records

Performance Criteria:

1.     Establish client needs and schedule analysis        
1.1    Liaise with client or sample provider to determine client needs and sample history
1.2    Record sample description, compare with specification and record and report discrepancies
1.3    Identify non-routine samples and the possible need to vary enterprise procedures
1.4    Seek advice from supervisor about any proposed variations and document all approved changes
1.5    Schedule analysis using enterprise procedures

2.     Prepare samples and standards        
2.1    Obtain a representative analytical portion of the laboratory sample
2.2    Prepare sample in accordance with testing requirements
2.3    Prepare validation checks for analytical portion(s)

3.     Set up and optimise instrument        
3.1    Perform pre-use and safety checks in accordance with enterprise procedures
3.2    Start up and condition the instrument using enterprise procedures
3.3    Optimise instrumental parameters to suit sample and test requirements
3.4    Check calibration status of instrument and perform calibration using specified standards and procedures, if applicable

4.     Perform analysis        
4.1    Measure analyte response for standards, validation checks and samples
4.2    Conduct sufficient measurements to obtain reliable data
4.3    Return instruments to standby or shutdown condition as required

5.     Process and analyse data        
5.1    Confirm data is the result of valid measurements
5.2    Perform required calculations and ensure results are consistent with standards or estimations and expectations
5.3    Record results with the appropriate accuracy, precision and units
5.4    Analyse trends in data and/or results and report "out of specification" or atypical results promptly to appropriate personnel
5.5    Troubleshoot analytical procedure or equipment problems which have led to atypical data or results

6.     Maintain a safe work environment        
6.1    Identify risks, hazards, safety equipment and control measures associated with sample handling, preparation and analytical method
6.2    Use personal protective equipment and safety procedures specified for test method and materials to be tested
6.3    Minimise the generation of wastes and environmental impacts
6.4    Ensure the safe disposal of laboratory wastes
6.5    Clean, care for and store equipment and consumables in accordance with enterprise procedures

7.     Maintain laboratory records        
7.1    Enter approved data and results into laboratory information management system
7.2    Maintain equipment logs in accordance with enterprise procedures
7.3    Maintain security and confidentiality of laboratory data and enterprise information
7.4    Communicate results to appropriate personnel.

Learning Outcomes

Details of Learning Activities

Learning activities for this course will include:
• Teacher directed face-to face delivery of lessons
• Whole class Discussion
• Pair/group discussion
• Small group workshops
• Online quizzes
•  Worksheets including Revision
• Laboratory experiments
• Record keeping of experiments
• Presentations
• Research activities

See the Teaching Schedule (below) for details.

Teaching Schedule

Date (Mon)WeekTheory (1 hr)Prac (4 hr)Assessment

1* Course OverviewSafety in the Chromatography Lab
Making solutions

 * Paper Chromatography Theory - concepts & calcs  

 2 Paper Chromatography Theory Paper Chromatography of food dyes  

 3 TLC Theory concepts & calcs Thin Layer Chromatography of Vegetable pigments Group 1 oral prac report*

 4 Gel Chromatography (Size Exclusion Chromatography (SEC)) Theory Gel Chromatography Group 2 oral prac report*

 Ion Exchange (IE) Chromatography Theory  Ion Exchange Chromatography  Group 3 oral prac report*

Affinity chromatography & Multistep chromatography - concepts

Case study: CSL 
 Multistep chromatography - Rainbow chromatography: isolating a mixture of food dyes 
Group 4 oral prac report*
CSL Online Quiz

 7 Revisionno prac class
Week of 31/8
 8 Exam Week
 Test 1 (theory)
  Test 1

 9 HPLC theory & calculations - peak area, retention time, internal standards

Hydrophobic interaction Chromatogrpahy (HIC)

Solid Phase Extraction (SPE)
Solution Making for HPLC
HIC extraction of GFP from
E. coli cells.

 10 HPLC - isocratic v’s gradient chromatography theory 1. HPLC of caffeine in drinks
 Group 5 oral prac report*
 Week of 21/9
  Student vacation  


 HPLC - isocratic v’s gradient chromatography theory        
 1. HPLC of caffeine in drinks -
    interpreting results

 12Example calcs for analgesics HPLC prac,  interpreting results, practising calcs & answering Q’s 
 HPLC of Analgesics  (Painkiller) Tablets

TLC of aspirin, caffeine & paracetamol
 Prac Report 2 - HPLC of analgesics
 13 Example calcs for analgesics HPLC prac,  interpreting results, practising calcs & answering Q’s  HPLC of Analgesics  (Painkiller) Tablets  interpreting results  

 GC theory & calculations
Internal standards
 GC @ Applied Chemistry
Gas Chromatography of Xylene
 GC Online quiz
 15 Revision  
 Week of 2/11

 16 Exam Week Test 2 (Theory)
Test 2
Submit Logbook
 Week of 9/11

 17 Alternative assessment (Supplementary exams) if required 
  * Oral practical report
= powerpoint presentation to class

Learning Resources

Prescribed Texts

Perform Routine Chromatographic Techniques Practical Manual (available online)

• J Crowe, T. Bradshaw, P. Monk (2006)
Chemistry for the Biosciences. The essential concepts.
pp 270-274 Oxford University Press, Oxford.

Available from the course Learning Hub (Blackboard) or directly from the RMIT Library’s eReserve collection & is available after login from the following link:

Also available in Carlton Library as Hardcopy : Call No. 540.078 C953

• Kaplan, Lawrence A., Pesce, Amadeo J. & Kazmierczak, Steven C. (2003)
Clinical chemistry: theory, analysis, correlation, 4th ed. St. Louis, Mo, Mosby
Chapters 5,6 & 7
Available from the course Learning Hub (Blackboard) or directly from the RMIT Library’s eReserve collection & is available after login from the following link:

The links to both e-texts can also be accessed from the Chromatography Learning Hub site in the External Links section:



Other Resources

Overview of Assessment

Assessments for this course typically consist of:
• Class activities
• Assignments
• Presentations
• Exam

Assessment Tasks

Self Assessment
1. Logbook
Week done: all
Topic:  Logbook - signed off by student each week lab class is attended**
Week due:  16
%:  satisfactory

Facilitated Assessment
1. Prac Report 1 - Oral group report (powerpoint presentation to class)
Week done: One group per week only
Topic:  for experiments done during weeks 2, 3, 4, 5, and 9 to be given on previous week’s experiment
Week due:  Group 1 - Week 3*/Group 2 - Week 4*/Group 3 - Week 5*/Group 4 - Week 6*/ Group 5 - Week 10*
%:  20%

2. Quiz 1
Week done: 6
Topic:  CSL Research Online Quiz
Week due:  9
%:  satisfactory

3. Exam 1
Week done: 8
Topic:  Chromatography (Paper, TLC, SEC, IEC, multistep)
%:  30%

4. Prac Report 2  (written group assignment)
Week done: 12/13
Topic:  HPLC of Analgesic (Painkiller) Tablets
Week due:  15
%:  20%

5. Quiz 2
Week done: 14
Topic:  GC quiz
Week due:  15
%:  10%

6. Exam 2
Week done: 16
Topic:  HPLC & GC only - shorter test
Week due:  n/a
%:  20%

7. Logbook
Week done: all
Requirement:  Logbook - signed off if considered competent** by teacher & recorded on central record/ Sign off each week lab class is attended**
Week due:  Submit completed logbook at end of week 16
%:  satisfactory

8. Attendance
Week done: all
Requirement:  80% attendance or more is required
Week due:  n/a
%:  satisfactory
Total 100%

*This is the week when the talk is to be given, ie. the topic of the presentation is in the week before the talk. For example, TLC of vegetable pigments is done in week 3 lab class, the talk on this topic will be held in week 4. Results are to be presented along with a short discussion of trends in the data and what the results tell you about the mixture being separated. Talks should be no more than 10 minutes long. All group members must present part of the talk.

** see logbook information for what is required.

* If you miss a prac report class you will only be eligible to write up/present a different prac if you provide a medical certificate.  For this to occur you need to discuss your situation with the teacher as soon as possible. Other reasons for missing a write up prac do not mean that you can write up a different report.If you miss an oral report presentation day you will be able to present it at the next class you attend as long as you provide a medical certificate.

In other words
* If you miss the experiment that you are to present as an oral presentation, you may present a different experiment as long as you provide a medical certificate
* Practical report 2 is due 2 weeks after the day that the practical is performed.
* Students must attend at least 80 % of practical classes to pass the course.
* Students must pass all of the assessment tasks to pass the course.
* ***Reports (powerpoint for oral presentation or word document for written report) need to be submitted online via blackboard or by email to the teacher if blackboard is not available. The online plagiarism statement given in blackboard must be included with the emailed assignment to show that the student agrees to these conditions***

Your logbook must be signed off after completion each week by both the student AND the teacher.
The teacher will also signoff your logbook on the class sheet as a central record.

The central record is the evidence record that is required to show that your logbook has been signed off by you, the student, and the teacher.

(If you miss the class you do not need to get the sheet signed off - the teacher will record this as an absence). YOU WILL BE CONSIDERED NYC* FOR THAT WEEK UNTIL THE CENTRAL RECORD IS SIGNED OFF or YOU ARE MARKED ABSENT.

*NYC = not yet competent

When is a prac performance considered "competent’ (ie when can a logbook be signed off)?
Logbooks will only be signed off when the student has followed and completed a method adequately, and is therefore considered competent for that experiment.

This is evidenced by:


1. Show date of analysis/results at top of page at start of experimental record in logbook,
2. Attach loose pages to logbook securely (use tape or staples),
3. Include raw and analysed results in logbook as appropriate to particular method (see lab manual)
4. Results considered in context of expected results by answering Questions 1-4 in logbook*
5. Clean up lab work area and equipment adequately
6. Student signed off logbook entry once record of results are completed and clean up done

*Questions 1-4
If the experiment is performed and completed, there are two possibilities for obtaining competent performance of the technique:
A. Results are within the expected range OR
B. Results are not within expected range AND a reasonable troubleshooting and reflection process has occurred.

To provide evidence for either A or B (above) answer the following questions in the logbook after each prac:
1. Were your results in the expected range? Yes/No
2. What were the key findings/results?
3. If no, why not - what is the most likely cause of the discrepancy?
4. What would you do differently if you had the opportunity to do the experiment again?

When is a prac performance considered "not yet competent’?
(ie when will a logbook not be signed off?)

A student is considered to be ’not yet competent’ for an experiment if:

* The Student doesn’t attend class long enough to complete the experiment
* The Student doesn’t finish the procedure in a reasonable time
(nominally by the time class finishes - excluding when there are logistic issues that mean few/no one in the class finishes).
* The student doesn’t contribute to the group effort adequately
* Results are a debacle - several errors/extensive lack of following SOP (method).
* The experiment is not conducted in a safe manner
* The lab area and equipment are not adequately cleaned up

In the case of a not yet competent result, it is noted that there is no opportunity to repeat that particular experiment due to scheduling of classes. In this case, to achieve competency other experiments during the course should be done to a ’competent’ level and the student should show a good amount of improvement as semester goes along.

As evidenced by...

* Observations in class by the teacher (record reason for NYC grade)
* The attitude and work ethic of the student while doing experiments is considered appropriate by the teacher

Assessment Matrix

Course Overview: Access Course Overview