new technologies addressing challenges in fruit juice … · new technologies addressing challenges...
TRANSCRIPT
©2008 Waters Corporation
New Technologies Addressing
Challenges in Fruit Juice Testing
Antonietta Gledhill
Waters Corporation
©2008 Waters Corporation 2
Waters CorporationA Brief Introduction
Founded in 1958
Number of Employees: >5,000
Operate in 27 countries
—11 Manufacturing Facilities
—Products available >50 countries
©2008 Waters Corporation 3
Waters CorporationTechnology Focus
©2008 Waters Corporation 4
Company’s Philosophy: Innovate
©2008 Waters Corporation 5
Company’s Philosophy: Innovate
©2008 Waters Corporation 6
Company’s Philosophy: Innovate
©2008 Waters Corporation 7
Application Focus
Biopharmaceuticals
Chemical materials
Clinical
Environmental
Food & Beverage
Pharmaceutical
Proteomics
©2008 Waters Corporation 8
Use of Analytical Tools to Support Business Decisions
Research & Innovation
Manufacture Quality
Food safety & QA
Established
products
Reformulated
products
New
products
Incoming
Material
Production
monitoring
Product
release
Breakthrough
innovation
Product
stability
Functional
benefit
©2008 Waters Corporation 9
Routine Analysis
Research & Innovation
Manufacture Quality
Food safety & QA
©2008 Waters Corporation 10
Routine Analysis Workflow
SAMPLE PREPARATION
Generic, L-L, SPE…
CHROMATOGRAPHY
Alliance, ACQUITY UPLC, GC
DETECTOR
UV, FLR, ELSD, MS
SOFTWARE
Empower, TargetLynx
SEPARATION CHEMISTRY
SunFire, XBridge, ACQUITY
©2008 Waters Corporation 11
Routine Analysis Separation Chemistry
BEH Amide Column (2009)
— Carbohydrates & Sugars
Detector: ELSD
* *
5 Food Sugars1
2
3
0.0
Minutes
2.0 4.0 6.0 8.0 10.0 12.0 14.0 16.0 18.0 20.0
4
5
Low Salt Retention = No Salt Interference
1) Fructose, 2) Glucose, 3) Sucrose, 4) Maltose, 5) Lactose (each 1mg/mL), * Salt Interferences
with Salts
©2008 Waters Corporation 12
Gain Switching
Routine Analysis Separation Chemistry
Minutes
0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0
Malto-tri, tetra, penta, hexa, hepta-ose
Double Chocolate Stout
Food Sugar Standard
Xylo
se
Arab
inose
Glu
cose
Malt
ose
Lacto
se
Fu
cose
Malt
otr
iose
Malt
ote
traose
Malt
op
en
taose
Malt
oh
exaose
Malt
oh
ep
taose
1.3µL injection volumeSamples: 100% Beer (No dilution or filtering)
©2008 Waters Corporation 13
Routine AnalysisLiquid Chromatography: Acquity UPLC®
Soft drinks
Example 1: Established HPLC methods
Speed Advantage
©2008 Waters Corporation 14
Routine AnalysisLiquid Chromatography: Acquity UPLC®
PolyphenolsVitamin C
Advantages: Resolution and Speed
Soft drinks
©2008 Waters Corporation 15
Routine AnalysisLiquid Chromatography: Acquity UPLC®
Benefits of Acquity UPLC
Chromatographic resolution
— Interferences / closely eluting
peaks
— …Selectivity from Core Detectors
Reduction in run-times
— Routine analysis
— Method development
Financial savings
— Solvent
— No of Instruments in lab
(servicing…)
©2008 Waters Corporation 16
Routine AnalysisDetectors - MS
ACQUITY SQD
Complementary tool for core detector
applications
Mass analysis for chromatographer
Selective & sensitive detector
IntelliStart with automated
— Set-up
— Calibration
— Method development
— Instrument diagnostics
©2008 Waters Corporation 17
Time0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50
AU
0.0
5.0e-3
1.0e-2
1.5e-2
2.0e-2
2.5e-2
3.0e-2
3.5e-2
4.0e-2
4.5e-2
5.0e-2
5.5e-2
6.0e-2
6.5e-2
7.0e-2
1.73
1.52
0.35
0.22
1.24
0.51
0.84
2.35
2.02
1.90
2.17
2.66
2.46
5.50
3.17
3.08
2.79
2.95
3.41
3.28
5.16
4.81
5.73
Routine AnalysisDetectors - MS
Additional security of selectivity & confirmation using MS
m/z100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500
%
0
100
%
0
100443
293
257
191
147134131121105
179173149 187
209193 256214
233229 247 264
277293
385
341294
323315299 311
329
355347 367 375
407386
406396 442408 433421
444
445
491475458 483 497
353
191
176
141
119109125 161
158173 179
192353201
198217 218 333233 319301269
239253 283 286
315
375
354
355
358
433411408
376 395431 451438
452463
491481 493
Advantage 1: Selectivity of MS in full scan mode
©2008 Waters Corporation 18
Routine AnalysisDetectors - MS
Time0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80
%
0
1.08
MS: m/z 271
Additional security of selectivity & confirmation using MS
Advantage 1: Selectivity of MS in SIR mode
©2008 Waters Corporation 19
Routine AnalysisDetectors – ELSD and MS
Stevia(5mg/mL)
Stevia Standards
Iso
ste
vio
l
Ste
vio
sid
e
Reb
au
dio
sid
e A
Reb
au
dio
sid
e C
Minutes0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0
0100200300400500600700800900
1000
ELSD (77.5% ACN with 0.2% TEA)
MS (77.5% ACN with 0.1% NH4OH) SIR of 5 Channels ES-TIC
1.15e6
Minutes
0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.00.0
%
0
100
m/z = 966.1(Rebaudioside A)
m/z = 950.1(Rebaudioside C)
m/z = 803.8(Stevioside)
Stevia(50µg/mL)
m/z = 318.5(Isosteviol)
Advantage 2: Sensitivity of MS
©2008 Waters Corporation 20
Summary for Routine Analysis
Technological Solutions to support Beverage Analysis
Separation Chemistries – Continual
research and development efforts here to
support challenging applications
ACQUITY UPLC – Excellent resolution &
increased speed of analysis
SQD MS – Additional confirmatory tool
benefitted from selectivity & sensitivity of
MS
©2008 Waters Corporation 21
Research:
Investigative / Troubleshooting
Research & Innovation
Manufacture Quality
Food safety & QA
©2008 Waters Corporation 22
Research: Investigative Analysis, Troubleshooting
Examples include:
New Product Development
— Ingredient / product profiling and characterisation
— Structural elucidation
— Degradation / transformation process
— Stability studies
Troubleshooting
— Authenticity and origin
— Manufacturing: Process optimisation
— Final Product: Contaminant screening
— Customer complaints & Competitor analysis
©2008 Waters Corporation 23
Research: Investigative Analysis…Published Data
“…To advance our understanding of the benefits
of GADF8 it is important to thoroughly
characterize its putatively active phenolic
components.
Mass spectrometry coupled to liquid
chromatography (LC/MS) has been increasingly
used in the structural characterization…”
©2008 Waters Corporation 24
Research: Investigative / Troubleshooting
SAMPLE PREPARATION
Generic, L-L, SPE
CHROMATOGRAPHY
ACQUITY UPLC, GC
DETECTOR
Tof (Time of flight) MS
SOFTWARE
ChromaLynx, MarkerLynx, MetaboLynx, MassFragment
SEPARATION CHEMISTRY
ACQUITY BEH, ACQUITY HSSGENERIC
METHOD
CONDITIONS
©2008 Waters Corporation 25
Research: Xevo QTof MS
Xevo QTof MS
DATA ACQUISITION
MODE
Low collision energy mode produces M+H or M-H
Ramp collision energy mode produce fragmentation data
©2008 Waters Corporation 26
Research: Software Tools
• Deconvolute Chromatograms: ID components in samples
• Sample Comparison: common & unique components
• Library search
QTof MS
DATA
• Multivariate statistical tool (PCA, PLS-DA)
• Perform database search (e.g. ChemSpider, user database) to ID potential markers
• Enables rapid metabolite identification (expected & unexpected)
• Integration with MassFragment for structural elucidation
• Structural Elucidation of observed exact mass ion fragments
©2008 Waters Corporation 27
Research: Investigative / Troubleshooting
SAMPLE PREPARATION
Generic, L-L, SPE
CHROMATOGRAPHY
ACQUITY UPLC, GC
DETECTOR
Tof (Time of flight) MS
SOFTWARE
ChromaLynx, MarkerLynx, MetaboLynx, MassFragment
SEPARATION CHEMISTRY
ACQUITY BEH, ACQUITY HSS
• Dilute and shoot
• ACQUITY column
• ACQUITY UPLC
• Xevo QTof MS (MSE mode)
• ChromaLynx XS• MassFragment
©2008 Waters Corporation 28
Research: Investigative / Troubleshooting
• Deconvolute Chromatograms: List components in sample
• Sample Comparison: Common & unique components
• Library search
©2008 Waters Corporation 29
Research: Investigative / Troubleshooting
[C16H11N2O4S]-
Precursor Information
©2008 Waters Corporation 30
Research: Elemental Composition & Chemspider
[C16H11N2O4S]-
Structural Elucidation Tools
©2008 Waters Corporation 31
Research: MassFragment
Product Ion Fragments
[C16H11N2O4S]-
Structural Elucidation Tool
©2008 Waters Corporation 32
Research:Investigative / Troubleshooting
SAMPLE PREPARATION
Generic, L-L, SPE
CHROMATOGRAPHY
ACQUITY UPLC, GC
DETECTOR
Tof (Time of flight) MS
SOFTWARE
ChromaLynx, MarkerLynx, MetaboLynx, MassFragment
SEPARATION CHEMISTRY
ACQUITY BEH, ACQUITY HSS
• Dilute and shoot
• ACQUITY BEH C18
• ACQUITY UPLC
• Xevo QTof MS (MSE mode)
• MarkerLynx• MassFragment
©2008 Waters Corporation 33
Research: MarkerLynx XS (Chemometric Software)
Sample List
Marker Table
S-Plot results
Trend-PlotChromatogram
©2008 Waters Corporation 34
Research:Chemometrics visual result
Kenya
Colombia
Colombia
Ethiopia
JavaBlend
Nicaragua
Costa Rica
©2008 Waters Corporation 35
Research: Grouping within a region
©2008 Waters Corporation 36
Research: Marker Identification
©2008 Waters Corporation 37
Research:Marker Database
©2008 Waters Corporation 38
Summary for Research Analysis
Technological Solutions to support Beverage Analysis
ACQUITY UPLC – Excellent resolution
and speed of analysis / method
development
Xevo QTof MS – Confirmatory exact
mass tool allowing experimental diversity
Software Tools – Provide intelligent
information to support investigative analysis
in research and troubleshooting environments
©2008 Waters Corporation 39
Thank-you for your attention
Any Questions?