RI-INBRE Workforce Development Training (WDT) Summer 2023 Syllabus
Module Title: Small Molecule Separation
Instructor: Ang Cai, PhD, and Sam Howarth
Course Overview
This 2.5-day RI-INBRE Workforce Development Training (WDT) Module provides an introduction to small molecules separation and chemical structure analysis methods. The module includes lectures on High Performance Liquid Chromatography (HPLC), Nuclear Magnetic Resonance (NMR), and Liquid Chromatography-Mass spectrometry (LC/MS) methods, as well as hands-on
laboratory experience with these techniques. This module is designed for students new to separation and analysis of small molecules and provides sufficient background to pursue further training in related research areas. Upon completion, participants will receive an RI-INBRE Certificate of Completion.
Learning Outcomes
- Develop kno0wledge of basic chemical separation techniques
- Small molecule sample preparation
- Proper operation of instrumentation including HPLC, NMR, and LC/MS
- Analysis of chemical structures
- Outcomes
Lab Report
Students will be expected to maintain detailed laboratory notes to include:
- Sample preparation
- Chemical analysis protocols
- Results
Resources Used in this Module:
- Ibuprofen
- Caffeine
- Aspirin (acetylsalicylic acid)
- Benzoic Acid
- Acetominophen (paracetamol)
The chosen small molecules are readily available from commercial sources. In addition, these
molecules have different chemical properties that will ensure successful HPLC separation. Each
compound is also commonly used in pharmaceutical research, making them relevant to a wide
range of scientific disciplines. Again, the structures of these molecules are well-known, which
will ensure straightforward structural characterization with NMR, as well as accurate
identification of their molecular weights using mass spectrometry.
Standard Operating Procedure – Thermo Ultimate 3000 HPLC
Standard Operating Procedure – Brucker 300 MHz NMR
Standard Operating Procedure – ABSciEX TOF 4600 or Thermo LTQ-XL Mass Spectrometer
Timeline
| Day 1 | Day 2 | Day 3 | |
|---|---|---|---|
| 9:00 AM-10:00 AM | Introduction to HPLC (Lecture) | Introduction to mass spectrometry and demonstration of how to use the instrument (Lecture) | Conclusion and discussion of the experiment results (Open Discussion) |
| 10:00 AM-11:00 AM | Preparation of samples for HPLC analysis (Hands-On) | Analysis of the separated compounds from LC with MS (Hands-On) | Conclusion and discussion of the experiment results (Open Discussion) |
| 11:00 AM-12:00 PM | HPLC Separation (Hands-On) | Analysis of the separated compounds from LC with MS (Hands-On) | Conclusion and discussion of the experiment results (Open Discussion) |
| 12:00 - 1:00 PM | Break | Break | Closing Group Lunch |
| 1:00 PM-2:00 PM | NMR Spectroscopy introduction (Lecture) | Interpretation of HPLC, NMR, and MS data to identify the two unknown compounds (Lecture/Hands- On) | |
| 2:00 PM-3:00 PM | Samples preparation, structure acquisition and Analysis (Hands- on) | Interpretation of HPLC, NMR, and MS data to identify the two unknown compounds (Lecture/Hands- On) | |
| 3:00 PM-4:00 PM | Review and discussion of HPLC and NMR data (Hands-on) | Interpretation of HPLC, NMR, and MS data to identify the two unknown compounds (Lecture/Hands- On) |