Course Title: Apply routine chromatographic techniques

Part B: Course Detail

Teaching Period: Term2 2013

Course Code: ONPS5663C

Course Title: Apply routine chromatographic techniques

School: 155T Vocational Health and Sciences

Campus: City Campus

Program: C5282 - Diploma of Laboratory Technology (Biotechnology)

Course Contact: Amberlee Mitton

Course Contact Phone: +61 3 9925 8053

Course Contact Email:

Name and Contact Details of All Other Relevant Staff

Kirsten Balding

contact by email, or talk in class

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

MSL974003A Perform chemical tests and procedures
MSL973007A Perform microscopic examination
MSL974006A Perform biological procedures
MSL974001A Prepare, standardise and use solutions

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:

MSL975009A Apply routine chromatographic techniques


1. Establish client needs and schedule analysis

Performance Criteria:

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

Performance Criteria:

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


3. Set up and optimise instrument

Performance Criteria:

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

Performance Criteria:

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

Performance Criteria:

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

Performance Criteria:

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

Performance Criteria:

7.1. Enter approved data and results into laboratory information management system (LIMS)
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 & theory activities
•  Worksheets including Revision
• Laboratory experiments
• Record keeping of experiments
• Presentations
• Research activities

See the Teaching Schedule (below) for details.

Teaching Schedule

Date (Mon) Week Theory (online) Prac (4 hr) Assessment
8/7 1 * Course Overview Safety in the Chromatography Lab
Making solutions
 Safety checks

  * Paper Chromatography Theory - concepts & calcs    
15/7  2  Paper Chromatography Theory  Paper Chromatography of food dyes  
22/7  3  TLC Theory concepts & calcs  Thin Layer Chromatography of Vegetable pigments  

Week 3 Quiz
29/7  4  Gel Chromatography (Size Exclusion Chromatography (SEC)) Theory  SEC Chromatography Group 1 oral prac report*
5/8 5  Ion Exchange (IE) Chromatography Theory  Ion Exchange Chromatography Week 5 Quiz Group 2 oral prac report
12/8 6 Affinity chromatography & Multistep chromatography - concepts

Case study: CSL 
 Multistep chromatography - Rainbow chromatography: isolating a mixture of food dyes

Week 6 Quiz -
CSL case study

Group 3 oral prac report*

19/8  7  Revision no prac class  
Week of 26/8  8  Exam Week  Test 1 (theory)   Test 1
2/9  9  HPLC theory & calculations - peak area, retention time, internal standards

Solid Phase Extraction (SPE)
Solution Making for HPLC
GFP &/or BFP from E. coli cells.

Week 9 Quiz

Group 4 oral prac report*

 9/9  10  HPLC - isocratic v’s gradient chromatography theory HPLC of caffeine in drinks Group 5 oral prac report*

Prac Report 2 - caffeine in drinks (experiment)
 16/9 11  HPLC - isocratic v’s gradient chromatography theory  HPLC of caffeine in drinks -
interpreting results
 Prac Report 2 - caffeine in drinks (analysis)
Week of  23/9  

Student vacation

 29/9  12 Example calcs for analgesics HPLC prac,  interpreting results, practising calcs & answering Q’s   HPLC of Analgesics  (Painkiller) Tablets

TLC of aspirin, caffeine & paracetamol
 Week 12 Quiz
7/10  13  Example calcs for analgesics HPLC prac,  interpreting results, practising calcs & answering Q’s  HPLC of Analgesics  (Painkiller) Tablets  interpreting results  Submit Logbook
14/10 14  GC theory & calculations
Internal standards
 GC @ Applied Chemistry
Gas Chromatography of Xylene
(to be confirmed)
Week 14 Quiz
 21/10  15  Revision    
 Week of 28/10
 16  Exam Week  Test 2 Test 2
 Week of 4/11
 17  Alternative assessment (special consideration only)    
     * 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 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


Other Resources

Overview of Assessment

Assessment for this course may include laboratory reports, assignments, exams and practival assessment in the laboratory

Assessment Tasks

1. Safety Checks
Week done: 1
Due: in class
%: satisfactory (ungraded)

2. Prac Report 1 - Oral group report (powerpoint presentation to class)
Week done: One group per week only at start of class
Topic:  for experiments done during weeks 2, 3, 4, 5, and 6
Week due:  Talk to be done 2 weeks after the experiment
%:  15%

3. Quizzes
Week done: 3,5,6,9,12,14
Topic:  see lab manual ’assessment guide’ section
Week due:  various - see blackboard
%:  20 (calculated from average grade)

4. Prac Report 2  (spreadsheet assignment)
Week done: 10/11
Topic:  HPLC of caffeine in drinks
Week due:  11 (in class activity)
%:  15%

5. Exam 1
Week done: 8
Topic:  Chromatography (Paper, TLC, SEC, IEC, multistep)
%:  25%

6. Exam 2
Week done: 16
Topic:  HPLC & GC only
Week due:  n/a
%:  25%

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 13 (monday week 14 is last day to avoid penalty to grades)
%:  satisfactory (ungraded)

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


Assessment Matrix

Other Information

*The topic of the presentation is 2 weeks 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 5. The presentation should contain a brief explanation of how the separation works, results and 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.


· 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 (a spreadsheet activity) is due in the analysis session.
· Each quiz has a published due date. Quizzes are to be done before the class in the same week to help you prepare for the class. Quizzes may include questions on safety.
· To pass the quizzes component you must get an average pass for all quizzes (ie. 50% average). The score for quizzes is calculated from the average grade of all quizzes. If you miss a quiz and have submitted a medical certificate you may do it later than the due date as long as you contact the teacher asap after your absence.
· 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 should be submitted online via blackboard.
· Please read the online anti-plagiarism statement given in blackboard submission of work via Blackboard incorporates student agreement to these conditions.


Course Overview: Access Course Overview