Upload
cpsa-20125-minutes-fame
View
336
Download
2
Tags:
Embed Size (px)
DESCRIPTION
5 minutes of fame
Citation preview
Agilent Restricted1
CE Capillary
N2Gas
Sheath Liquid
Microsampling/DBSLatest Innovations from Agilent Technologies
CE-MS Chip-MS DBS-MS
Pre Extraction Image
April 13, 2023
New Polaris-Chip for highest resolution
Stationary Phase
Enrichment column
Sprayer-tip
Analytical column
Inert polyimide
• Polaris C18-A, polar embedded phase,
3 µm, 180 Å, 200 m2/g
• 150 mm x 75 µm ID
• 1 µg sample loading
Loading capacity for new Polaris Chip
• Shown: E.coli TIC and EIC‘s of 500 ng injection superimposed
• Determined using 9 representative peptides across elution range
2x10
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
+ESI EIC(858.7801) Scan Frag=175.0V Load_500.d
Counts (%) vs. Acquisition Time (min)6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40
P1 P2 P3 P4 P5 P6 P7 P8 P9
April 13, 2023
Agilent CE-ESI-MS Setup
MassHunter SW control:
- CE instrument- MS instrument- LC makeup flows
Page 5
Small anionic molecules in Metabolomics by CE-ToF-MS
CE-MS is ideally suited to handle ionic or polar metabolites in complex mixtures reproducibly
04/13/2023CPSA Shanghai
6
Dried Blood Spots (DBS) Analysis
• µLs sample• Stable on card at ambient temp• Easy to ship• Non-biohazardous
• mLs sample• Must store frozen• Difficult to ship• Biohazardous
04/13/2023CPSA Shanghai
7
An Automated Online Card Extraction
LC/MS System for Dried Blood Spots
Analysis
DBS CardRacks
Robotic Gripper
SwitchingValvesClamp Module
Camera
ISTD Dilutor
04/13/2023CPSA Shanghai
8
Agilent Automated Card Extraction (AACE) LC/MS System Flow Schematic
Clamped DBS Card
10-PortValve
6-PortValve
Extraction Pump (1)1260 Binary LC
Dilution Pump1200 Isocratic LC
Analytical Pump (2)1260 Binary LC
ISTD
ISTDLoop
Trap 1 Column
Trap 2 Column
Analytical Column
A B A A BWaste
6460 QQQMaster Control
Prolab Card Extration Instrument
Agilent Restricted9
MRM chromatograms of clozapine, noreclozapine, and clozapine-N-oxide
Reproducible and sensitive DBS/MS systemLOQs for all compound 0.5 ng/mL in blood
Clozapine Norclozapine Clozapine-N-oxide
04/13/2023CPSA Shanghai
10
Summary: AACE System vs. Offline DBSComparable Quantitation Performance Parameters
AACE LC/MS System
Offline Extraction DBS
CompoundLOQ
(ng/mL in blood)Range
(ng/mL)Linearity
(R2)Accuracy
(%)Reproducibility
(%)Precision
(%)
Clozapine 0.5 0.5 – 10000 0.9996 92.1 – 106.8 0.61 – 5.61 4.82
Norclozapine 0.5 0.5 – 5000 0.9988 85.2 – 109.9 0.84 – 6.05 8.94
Clozapine N-oxide 0.5 0.5 - 5000 0.9981 93.2 – 104.1 0.68 – 6.15 3.82
Analysis Time ~ 1 hr
CompoundLOQ
(ng/mL in blood)Range
(ng/mL)Linearity
(R2)Accuracy
(%)Reproducibility
(%)Precision
(%)
Clozapine 0.5 0.5 – 10000 0.9997 88.0 – 103.2 0.61 – 8.92 4.89
Norclozapine 0.5 0.5 – 10000 0.9991 89.3 – 107.1 0.08 – 8.98 8.36
Clozapine N-oxide 0.5 0.5 - 10000 0.9991 91.8 – 108.4 0.25 – 8.66 6.64
Analysis Time ~ 5 hrs
04/13/2023CPSA Shanghai
11
DBS Summary – fast and “fit for purpose”
Agilent Automated Card Extraction (AACE) • Automated flow-through DBS analysis • Fully integrated with MassHunter LC/MS
Data Processing and Reporting• Pre- and post- sampling DBS card images• Processed quantitative data• Chain of custody
Clozapine study • Sensitivity: 0.5 ng/mL LLOQ • Linear assay over 4 orders of magnitude• Comparable results with offline method• Time savingss, 1 hour versus 5 hours