LC–MS Workflows — HILIC RP & Ion Modes — Hands-on

Build practical LC–MS metabolomics skills from a method-development perspective. Learn how to choose between RP and HILIC, design gradients and buffers, optimize ionization, and manage positive/negative polarity modes. This module helps you create reproducible LC–MS methods that integrate cleanly with downstream peak picking, alignment and quantitation workflows.

LC–MS Workflows — HILIC RP & Ion Modes

Help Desk · WhatsApp

Session Index

Session 1 — LC–MS Principles & Components Session 2 — RP LC–MS Method Development Session 3 — HILIC & Ion Source / Polarity Strategies Session 4 — Mini Capstone: LC–MS Method & Injection Plan

Session 1

Fee: Rs 8800

LC–MS Principles & Components

Liquid chromatography for metabolomics: overview

analytical vs UHPLC systems pumps, autosamplers, columns backpressure and dwell volume

Mass spectrometer overview for LC–MS

quadrupole, TOF, Orbitrap basics resolution and dynamic range MS1 vs MS/MS acquisition

Connecting LC and MS: interfaces & plumbing

ESI source layout capillaries and fittings dead volume and peak broadening

Session 2

Fee: Rs 11800

RP LC–MS Method Development

Reversed-phase principles for small molecules

C18 and alternative chemistries retention vs hydrophobicity column temperature control

Mobile phases, buffers and gradients (RP)

water / acetonitrile / methanol formic acid, ammonium formate/acetate gradient shapes and re-equilibration

Injection parameters and carryover control

injection volume and solvent strength needle washes and blanks column conditioning and stability

Session 3

Fee: Rs 14800

HILIC & Ion Source / Polarity Strategies

HILIC principles and method setup

polar stationary phases high organic starting conditions longer column equilibration

Ionization source tuning and parameters

ESI voltage and temperature gas flows and nebulization minimizing adducts and in-source fragments

Polarity and ion-mode strategies

positive vs negative ion mode polarity switching considerations coverage vs duty cycle trade-offs

Session 4

Fee: Rs 18800

Mini Capstone: LC–MS Method & Injection Plan

Choosing RP vs HILIC for a use case

clinical, microbiome or cell culture study

Designing gradient, source and polarity settings

mobile phase and buffer recipe source tuning notes single vs dual polarity runs

Injection order, QC layout and method checklist

blanks, QCs and standards system suitability tests method & maintenance log template

PDF

LC–MS Workflows — HILIC RP & Ion Modes Training | LC–MS Methods, Chromatography & Ionization

LC–MS Workflows — HILIC RP & Ion Modes — Hands-on