acquity uplc & xevo tq-s (waters) - wur · xevo tq-s acquity uplc accurate and sensitive...
TRANSCRIPT
UPLC-triple Quad for targeted quantitation
Acquity UPLC & Xevo TQ-S (Waters)
Plant Sciences group & RIKILT
Xevo TQ-S
Acquity UPLC
Accurate and sensitive detection and quantitation of small biomolecules, such as:
• Metabolites, Hormones
• Toxins, elicitors
• Drugs, pesticides
• Peptides (proteins)
specific expertise for:
• development of targeted methods for protein quantitation based on proteomics marker analyses
• targeted detection of plant hormones
SRM / MRM principle
[Protein digest]
SPE sample prep
LC-MS separation
Precursor MS/MS
Fragment m/z
SRM transition
● Q1: precursor m/z
● Q2: fragmentation
● Q3: fragment m/z
Target extract SPE
Proteomics MS: peptide quantification
Compounds/Peptides elute from LC elution profile
MRM signal is integrated over peak area
Peak area is directly correlated to peptide load
Relative quantitation between samples
Synthetic peptides as (quantitative) reference
Absolute quantitation possible
Keywords
Targeted detection
● Selective
● Sensitive
● Quantitative (accurately)
● Broad concentration range
● Multiplexed
● Diverse
● Fast
Targets: Metabolites Hormones Toxins, elicitors Drugs, pesticides Peptides (proteins)
Method comparison
Untargeted LC-MSE
● Identification and (semi)-quantitation of a large number
of proteins in complex samples: thousands of peaks
● Optimise separation and resolution( long runs, ~2 /day)
● Highly detailed quantitative comparison
● Determine optimal marker peaks & fragments
● Transfer targeted method to QQQ
Targeted Triple Quadrupole MS (QQQ)
● Selected set of peptides
● Increased separation speed (typically 10min.)
● Increased sample throughput (up to 150/day)
LC-MS for identification and quantitation
