Waters white paper Solution 2.3.1. Waters HPLC and Fluorescence Detector 2.3.2. Waters Quadrupole...
32
[ WHITE PAPER ] TOTAL SOLUTIONS FOR ENVIRONMENTAL APPLICATIONS
Transcript of Waters white paper Solution 2.3.1. Waters HPLC and Fluorescence Detector 2.3.2. Waters Quadrupole...
[ white paper ]
total solutions for environmental applications
1. total solut ions for env ironmental ap pl icat ions 3
2. carbamat es 3
2.1.IntroductionofCarbamatePesticides
2.2.MethodofAnalysis
2.3.WatersSolution
2.3.1.WatersHPLCandFluorescenceDetector
2.3.2.WatersQuadrupoleMassSpectrometerwithHPLC
2.3.3.GPCCleanupSystemandEnvirogelColumn
3. polynuclear aromat ic Hydrocarbons (paHs) 7
3.1.IntroductionofPAHs
3.2.EPAMethodsfortheAnalysisofPAHs
3.3.WatersSolution
3.3.1.WatersBreezeSystem
3.3.2.WatersAllianceSystem
3.3.3.WatersACQUITYUPLCSystem
a.withTUVdetector
b.withFluorescencedetector
4. endoc rine disru ptors (edc s) 13
4.1.IntroductionofEndocrineDisruptors
4.1.1.WhatareEndocrineDisruptors?
4.1.2.AnalysisofEmergingContaminantsinWastewater
4.1.3.AnalysisofEDCsinDrinkingWater
4.2.WatersSolution
4.2.1.PharmaceuticalCompoundsintheEnvironment;AnalysisofsurfaceandgroundwaterbyLC/MS/MS
4.2.2.PharmaceuticalCompoundsintheEnvironment;ConfirmationofresiduesbyLC/Q-Tof
4.2.3.AConfirmatoryMethodfortheDeterminationofSyntheticPyretheticinWastewater
5. formaldeHyde analysis 20
5.1.IntroductionofFormaldehyde
5.2.WatersSolution
5.2.1. HPLCAnalysis
5.2.2. WatersACQUITYMethodforFormaldehyde
6. dioxins 24
6.1.IntroductionofDioxins
6.2.USEPAMethod1613
6.3.WatersSolution
6.3.1.DioxinScreeningusingGC/MS/MS(QuattromicroGC)
6.3.2.DioxinConfirmationusingHRMS(AutoSpecPremier)
7. Wat ers sof t Ware solut ions 30
7.1.MassLynxRelatedMSProducts
7.2.EmpowerSoftware
8. Wat ers Global serv ic es 31
3
1. total solut ions for env ironmental ap plicat ions
introduction
The environmental segment is a complex market consisting of a
numberofinstrumentsappliedtoalltypesofmedia.Environmental
instrumentationisprimarilyusedtodetect,monitorandmeasure
the presence of hazardous or potentially hazardous chemicals in
water,soilandair.
Themajorityof analysesperformed in this fieldarequantitative.
Screeningmethodscanbeusedtoidentifyandeliminatesamples
containing high concentrations of target analyte(s). Remaining
samples can thenbe transferred tomore selective techniques to
quantifyandconfirmlowlevelanalytes.Thereisageneraldown-
wardtrendinregulatoryreportinglevels,requiringmoresensitive
instrumentations.
Environmentalcontrolisaregulatedapplication,havinglegaland
eventually financial implications upon the results. Although the
regulationsarenotasstrictastheyareforfoodcontrol,thereis
anobligationtohaveasystemcapableofanalyticalqualitycontrol
thatissubjecttochecksbyapersonwhoisnotunderthecontrol
ofthelaboratoryandwhoisapprovedbythecompetentauthority
forthatpurpose.
in that context, the following are especially important:
n Robustnessandreliabilityofanalyticaltechnique
n Instrumentdown-time
n Frequencyofmaintenance
n Easeofmaintenance
n Rapidserviceresponsewhennecessary
n Sensitivity:asensitiveinstrumentneedsless
samplepreconcentration
n Samplethroughput
n Selectivity:aninstrumentormethodthatisabletoprovide
highresolution
n Globalcostofuse.Environmentalcontrolisahighlycompeti-
tivebusiness.Contractlaboratoriesareoftenincompetition
withuniversitiesandpubliclaboratories,whocanoperate
withminimizedpersonalandinstrumentcosts.Thesestudies
involvecostofequipment,staffingcosts(training,working
hours),costofsamplepreparationandsamplethroughput.
Thatiswhyeaseofoperation,automation,minimalsample
treatmentandanalysisspeedareimportantwhenchoosing
atechnique.
2. carbamat es
2.1. introduction of carbamate pesticides
Carbamate, thiocarbamate and urea-based pesticides are com-
monlyusedasagriculturalpesticides.
The field run-off water transports these analytes into the soil,
ground water and tributaries. Waste treatment does not remove
allpesticidesbeforetheyaredischargedintothetributaries.These
arethesourcesofourdrinkingwater.Pesticideshavedemonstrated
toxicologicaleffectsintheenvironmentandhavebeenimplicated
asendocrinedisruptors.
Thereisastrongneedforanalyticalmethodstoscreenandquantify
carbamatesinrawagriculturalcommodities,surfaceanddrinking
water,andsoil.
u.s. epa regulation on carbamates
The United States Environmental Protection Agency (U.S. EPA)
is required to monitor the quality of drinking water and raw
sourcewater,soilandwastematricesroutinely.TheEPAmethods
include 531.2 for drinking water and 8318 for soil and waste
matrices. Thesemethods requirepost columnderivatizationwith
fluorescence detection. Currently, EPA method 531.2 is used to
regulate two compounds carbofuran and oxamyl. The Maximum
Containment Level (MCL) for carbofuran and oxamyl in drinking
wateris0.04mg/Land0.2mg/L,respectively.
eu regulation on pesticides
EuropeanUnion(EU)regulationsfordrinkingwaterarespecified
intheECDirective98/83/EC.Themaximumadmissibleconcentra-
tionforpesticidesis0.1μg/Lforeachindividualcomponent,total
concentration0.5μg/L.
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2.2. method of analysis
LiquidChromatography(LC)isthepreferredseparationtechnique
for carbamates, thiocarbamates and phenylurea because most of
themarepolarandthermallylabile. InGasChromatography(GC)
analysis,thesecompoundseithershowsignsofthermaldecompo-
sitionorfailtoelutefromthecolumn.
There are three approaches to the analysis of carbamates. The first
approachisusingLCwithaUVdetector.Althoughthismethodissimple
anddirect,itcanonlydetectcompoundsthathaveUVabsorbance.In
addition,itrequiresanotheranalyticalmethodforconfirmation.
ThesecondapproachisusingLCwithfluorescencedetection,which
is a more sensitive method than UV. Compounds that are not UV
activecanbederivatizedtogivefluorescenceproperties.However,
likeUVdetection,itrequiresasecondarymethodforconfirmation.
ThethirdapproachisusingLC/MSorLC/MS/MS.Compoundsthatare
neitherUV-norfluorescence-activecannotbedetectedbyEPAmeth-
ods531.2and8318.Massspectrometryofferstheabilitytoanalyzea
largerdiversityofcompoundswithsufficientsensitivityandselectivity
thatprovidesaone-stepapproachforanalysisandconfirmation.
major considerations with different analytical methods
instrument derivation resolutionsecondary
metHod
larGer ranGe of
compoundssensitivity selectivity
HPLC and Fluorescence
detectorYes Yes Yes No Good Good
LC/MS No No No Yes Very good Very ood
LC/MS/MS No No No Yes Excellent Excellent
Waters application notes/Journals
1.“A Fully Automated LC/MS Method Development and Quantification ProtocolTargeting 52 Carbamates Thiocarbamates and Phenylureas.” AnalyticalChemistry.200375:4101-412.
2.“LC/MSQuantificationof52Carbamates:AFullyAutomatedProtocol.”WatersCorporation,2002:720000176EN.
3.“A Fully Automatic Multi-Analyte Quantification Protocol for Carbamates – AComparisonofLC/MSVSLC/MS/MS.”WatersCorporation,2007:720000672EN.
structures of carbamates
CH3 S C CH N OR
O CH3
CH3
1. Aldicarb sulfoxide
CH3 S C CH N OR
O CH3
CH3O
2. Aldicarb sulfone
(CH3 )2 C C N OR
O
SCH 3
N
3. Oxamyl
CH3 C N OR
SCH3
4. Methomyl
ORHO
CH3 CH3
5. 3-Hydroxycarbofuran
CH3 S C CH N OR
CH3
CH3
6. Alidcarb
OR
OCH3 CH3
7. Propoxur
OR
OCH3 CH3
8. Carbofuran
OR
9. Carbaryl
OH
10. 1-Naphthol
CH3S OR
CH3
CH3
11. Methiocarb
O
Where R =
C NH CH3
O
R-O-C- NH-CH3
O
CH 3 NH 2 + R-OH + H2CO3
CH3NH2
HSCH2CH2OH
C-H
C-H
O
O
N-CH3
SCH2CH2OH
Aq. alkali
+
+
Aq. alkali
N-Methylcarbamate Methylamine
O-Phthalaldehyde (OPA)
2-MercaptoethonalHighly fluorescent derivatives 339 nm ex / 445 nm em
CH3 S C CH N OR
O CH3
CH3
1. Aldicarb sulfoxide
CH3 S C CH N OR
O CH3
CH3O
2. Aldicarb sulfone
(CH3 )2 C C N OR
O
SCH 3
N
3. Oxamyl
CH3 C N OR
SCH3
4. Methomyl
ORHO
CH3 CH3
5. 3-Hydroxycarbofuran
CH3 S C CH N OR
CH3
CH3
6. Alidcarb
OR
OCH3 CH3
7. Propoxur
OR
OCH3 CH3
8. Carbofuran
OR
9. Carbaryl
OH
10. 1-Naphthol
CH3S OR
CH3
CH3
11. Methiocarb
O
Where R =
C NH CH3
O
R-O-C- NH-CH3
O
CH 3 NH 2 + R-OH + H2CO3
CH3NH2
HSCH2CH2OH
C-H
C-H
O
O
N-CH3
SCH2CH2OH
Aq. alkali
+
+
Aq. alkali
N-Methylcarbamate Methylamine
O-Phthalaldehyde (OPA)
2-MercaptoethonalHighly fluorescent derivatives 339 nm ex / 445 nm em
Step 1: Hydrolysis.
Step 2. Derivatization of methyalmine.
Compound without the –NH-R group can not be derivatized, and cannot be analyzed by the post-column fluorescence method.
CH3 S C CH N OR
O CH3
CH3
1. Aldicarb sulfoxide
CH3 S C CH N OR
O CH3
CH3O
2. Aldicarb sulfone
(CH3 )2 C C N OR
O
SCH 3
N
3. Oxamyl
CH3 C N OR
SCH3
4. Methomyl
ORHO
CH3 CH3
5. 3-Hydroxycarbofuran
CH3 S C CH N OR
CH3
CH3
6. Alidcarb
OR
OCH3 CH3
7. Propoxur
OR
OCH3 CH3
8. Carbofuran
OR
9. Carbaryl
OH
10. 1-Naphthol
CH3S OR
CH3
CH3
11. Methiocarb
O
Where R =
C NH CH3
O
R-O-C- NH-CH3
O
CH 3 NH 2 + R-OH + H2CO3
CH3NH2
HSCH2CH2OH
C-H
C-H
O
O
N-CH3
SCH2CH2OH
Aq. alkali
+
+
Aq. alkali
N-Methylcarbamate Methylamine
O-Phthalaldehyde (OPA)
2-MercaptoethonalHighly fluorescent derivatives 339 nm ex / 445 nm em
5
2.3. Waters solutions
2.3.1. Waters® Hplc and fluorescence detector
chromatographic conditions:
Instrument: Alliance®systemforcarbamateanalysis
Column: Carbamateanalysis,3.9mmIDx150mm
Mobilephase:A-water,B-methanolC-acetonitrile
time %a %b %c curve
0.00 88 12 0 -
5.30 88 12 0 1
5.40 68 16 16 5
14.00 68 16 16 3
16.10 50 25 25 7
20.00 50 25 25 6
22..00 88 12 0 5
Temperature: 30°C
Flowrate: 1.5mL/min
Sample: 10ngofeachanalyteoncolumn
Injection: 400μL
Postcolumnreagent1: NaOHat0.5mL/min
Postcolumnreagent2: Orthophthaladehyde(OPA)/
2-Mercaptoethanolat0.5mL/min
Detection: Excitation:339nm,Emission:445nm
epa method 531.2 analytes
Minutes2.00 6.00 10.00 14.00 18.00 22.00
1
34
5
6
78
9
10
2
11
12
1500 E
U
Alliance HPLC system for carbamate analysis consists of a 2695 Separations Module with column heater assembly, post-column reaction module, temperature control module, two reagent man-gers, 2475 Scanning Fluorescence detector, Empower software and Waters’ carbamate analysis column.
peak analyte ppb
1 Aldicarb sulfoxide 25
2 Aldicarb sulfone 25
3 Oxamyl 25
4 Methomyl 25
5 3-Hydroxycarbofuran 25
6 Aldicarb 25
peak analyte ppb
7 Propoxur 25
8 Carbofuran 25
9 Carbaryl 25
10 1-Naphthol 25
11 Methiocarb 25
12 BDMC 25
6
2.3.2. Waters Quadrupole mass spectrometer with Hplc
chromatographic conditions:
Instrument: AllianceHT2795SeparationModulewith
MassLynx™software,2996PhotoDiodeArray
detectorandMicromass®ZQ2000massdetector
Column: Symmetry®C83.5um2.1mmIDx150mm
Sample: 50pgofeachanalyteoncolumn
Mobilephase:A-10mMNH4OAcinwater,pH5.0,
B-10mMNH4OAcinacetonitrile
binary Gradient:
time %a %b curve
0.00 95.0 5.0 1
40.0 30.0 70.0 6
50.0 0.0 100.0 1
64.0 95.0 5.0 1
Temperature: 40°C
Flowrate: 0.3mL/min
Injection: 50μL
ms conditions:
Ionization: ESI+
Capillaryvoltage: 3.5kv
Sourcetemperature: 140°C
Desolvationtemperature: 350°C
Desolvationgasflow(L/Hr): 650
Conegasflow(L/Hr): 0
LMresolution: 14.5
HMresolution: 14.5
Ionenergy: 1.5
Dwelltime(s): 0.02
Interchanneldelay(s): 0.02
Interscandelay(s): 0.02
%
%
28.59
17.71
16.959.07
5.22 7.78
5.46
16.72
11.75 12.14
15.71
20.86
20.46
17.96
19.03
26.86
24.8422.5122.46
23.12
25.86
27.82
28.64
32.50
31.4730.18
37.9835.03
33.2437.16 38.35
40.94
23.1
17.9912.15
11.74
9.097.75
6.69
16.95
16.72
13.15
22.50
21.54
20.8719.05
28.59
24.80 27.8427.57
30.22
32.5231.42
38.00
35.90 38.39
28.59
25.86
28.64
.
Time
2: Diode Array240
1 ppb100
100
8.00 12.00 16.00 20.00 24.00 28.00 32.00 36.00 40.00
1: Scan ES+TIC
Figure 1. Full Scan TIC of 38 carbamates.
Waters Alliance system with the 2996 PDA detector, Micromass ZQ mass detector and MassLynx software.
7
2.3.3. Gpc cleanup system and envirogel column
The Envirogel™ high-efficiency GPC cleanup columns are spe-
cificallydesignedtoremovelowvolatility,highmolecularweight
interferences,suchaslipidsandnaturalresins,fromenvironmental
samplesasspecifiedinEPAmethod3640A:
n Removeslow-volatilitycompounds
n MandatoryinCLPwork
n Semi-volatiles
n Pesticides
3. polynuclear aromat ic Hydrocarbons (paHs)
3.1. introduction of paHs
PAHsareagroupofmoleculesthataremalodorous,andmadeupof
manyringsofonlyhydrogenandcarbon.Therearetwocharacter-
isticsthatdefinethedifferenttypesofPAHmolecules:howmany
ringsithas;andhowthoseringsarearranged.
number of rings:SomeofthemorecommonPAHmoleculeshave
two,three,orfourrings.
shape of molecules:PAHmoleculeswithmorethantworingscan
have the rings arranged in different ways. The rings in the mol-
ecules can be in a straight line, bent in a curve, or compressed
together inablock.Benzo(a)pyrene,or“BaP”forshort, isoneof
mostdangerousPAHmoleculesasitiswell-knowncarcinogen.
0.00 5.00 10.00 15.00 20.00 25.00 30.00
0.00
500.00
1000.00
1500.00
2000.00
AU
Minutes
1 2 3
4
5
Columns: Envirogel 19mm x (30cm + 15cm)Mobile Phase: 100% Methylene Chloride at 5 mL/minDetection: UV at 254 nmSample Load: Compound 2 mL 5 mL
1. Corn Oil 62.5 mg/mL 25.0 mg/mL2. Phthalate 2.5 mg/mL 1.0 mg/mL3. Methoxychlor 0.5 mg/mL 0.2 mg/mL4. Perylene 0.05 mg/mL 0.02 mg/mL5. Sulfur 0.2 mg/mL 0.08 mg/mL
2 mL
5 mL
COLLECT
GPC Cleanup 2 mL vs 5 mL injection.
Naphthalene
Anthracene
Phenanthrene
Benzo(a)pyrene
Breeze GPC cleanup system.
GPC Cleanup system with Envirogel column.
8
summary:
2 rings: 3 rings:
Naphtalene Phenanthrene
Acenaphtylene Anthracene
Acenaphtene Fluoranthene
Fluoren Benzo(c)fluranthene
4 rings: 5 rings:
Pyrene Benzo(a)pyrene
Benzo(a)anthracene Bibenzo(a,h)anthracene
Benzo(b)anthracene Benzo(g,h,l)perylene
Indeno(1,2,3-cd)pyrene Benzo(k)fluranthene
TherearemanywayshumanactivitiesincreasethereleaseofPAHs
intotheenvironment.Themostcommoninstancesare:
oil spills:OilcontainsPAHs.Whenoilispipedupfromtheground,
spillsmayoccurresultinginPAHsbeingmovedfromdeepwithin
thesoilandreleasedontothesurface.Oilspillsalsocomeindif-
ferent forms, such as spills from the sinking or collisions of oil
tankers,orthedumpingofusedmotoroilbyautomechanics.These
directlycontributetoPAHpollutionofwaterandoil.
car exhaust:Whencarsortrucksburngasoline,PAHmoleculesare
releasedfromthecarexhaust.Thesameappliestodieselburning
trucks.Haveyou ever beenbehind a truck at a traffic light? The
cloud of black effluent released from the exhaust pipe when the
truckmovesoffisfullofPAHmolecules.
soot & smoke: When we burn wood, charcoal, or coal, we make
ashandsoot.BlacksootcontainsmanyPAHmolecules.Bigpower
plantsthatburncoalproducelotsofsoot.Eventhoughthebarbe-
cuegrillsattheparkmakeminisculeamounts–botharesourcesof
PAHpollution.PAHsthatcomefromsmokestacksalsocontribute
toPAHpollutionoftheair,whereasPAHsfromfireplacesandgrills
willcontributetothePAHpollutionofsoil.
method of analysis
PAHs,whichresultfromtheincompletecombustionoffossilfuels
and organic matter, are among the most frequently monitored
environmentalcontaminants.Becauseoftheircarcinogenicnature,
many of these methods specify HPLC, usually with fluorescence
and/orUVdetectionastherecommendedanalyticalprocedure.
Subpartsperbillion(ppb)detectionlimitsforthetargetanalytes
arenecessary,whichmeansthatthesamplepreparationrequiresa
pre-concentrationstep(EPAMethods610&8310.).Differentcon-
figurationsbasedontheWatersBreeze™binarypumpsorAlliance
Separationmodulescombinedwitha2475Fluorescencedetector
canbeusedtorunthiskindofanalysis.
The 2996 PDA may be used as a possible alternative when UV
spectrum identification is required. Separations are run on dedi-
catedC18columns.
eu regulation on paHs
Benzo(b)fluoranthene, benzo(k)fluoranthene, benzo(ghi)perylene,
andindenol(1,2,3-cd)pyreneareregulatedbytheEuropeanUnion
(EU)andtheirtotalsafetyconcentrationslimitfordrinkingwater
issetat0.10μg/L.
Naphthalene
Anthracene
Phenanthrene
Benzo(a)pyrene
u.s. epa regulation on paHs
The U.S. Environmental Protection Agency (EPA), and European
administrationregulatetheconcentrationofPAHsinsoils,drink-
ing water, wastewater, food, and solid waste. Currently, the EPA
regulates Benzo(a)pyrene in drinking water, and the maximum
contaminantlevel(MCL)is0.0002mg/L.
�
3.2. epa methods for the analysis of paHs
sample matrix sample preparation Hplc anaysis
TO-13 Ambient air Soxhlet extraction C18 column 40% -100%CH3CN linear gradient
550.1Drinking water
Table waterSuckling water
Extract with C18 solid phasecartridge elute with MeCl2
C18 column 30% -100%CH3OH linear gradient
610Waste water
Underground waterSurface water
Liquid/liquid extractioncartridge elute with MeCl2
C18 column 40% -100%CH3OH linear gradient
8310 Solid sedimentsSludge waste MeCl2 extraction C18 Column 40% -100%
CH3OH linear gradient
Waters application notes
1.Waters Breeze HPLC System, High Performance and Application Versatility,PerformancePerspective-720000373EN.
2.TheScienceofACQUITYUPLC®AppliedtoEnvironmentalAnalysesofPAHandExplosivesinWater-720001398EN.
3.3. Waters solution
3.3.1. Waters breeze system
chromatographic conditions:
Column: PAHcolumn,3.0mmx250mm,5μm
Mobilephase: Waterandacetonitrile
Flowrate: 0.5mL/min
Columntemperature: 30°C
Sample: EPA610mixture(1/10000dilution)
Injectionvolume: 5μL
12
3
4 5
6
7
8
9
1011
1213
14
15 16
UV at 254 nmBreeze system
6.00 10.00 14.00
Minutes
22.00 26.0018.00
Figure 2. EPA 610 standard mix of PAHs on Breeze HPLC system with UV detection.
peak analyte ppm
1 Napthalene 20
2 Acenaphthylene 40
3 Acenaphthene 20
4 Fluorene 4
5 Phenanthrene 2
6 Anthracene 2
7 Fluoranthene 4
8 Pyrene 2
peak analyte ppm
9 Benzo(a)anthracene 2
10 Chrysene 2
11 Benzo(b)fluoranthene 4
12 Benzo(k)fluoranthene 2
13 Benzo(a)pyrene 2
14 Dibenzo(a,h)anthracene 4
15 Benzo(g,h,l)perylene 4
16 Indeno (1,2,3-cd) pyrene 2
10
3.3.2. Waters alliance system
chromatographic conditions:
Instrument: AllianceHPLCsystemwithEmpower™2software,
2996PhotoDiodeArraydetector,2475
Fluorescencedetector(connectedinseries)
Column: PAH(4.6x250mm,5μm)
Temperature: 27°C
EluentA: Water
EluentB: Acetonitrile
Gradient: 60%Bto100%B,curve9,for12mins,Holdat
100%Bfor11mins,returntoinitialconditions
Flowrate: 1.2mL/min
UVdetection:Scan200to350nm,extractedat254nm
Fluorescence:Time-programmedwavelengthchanges
Sample: EPA610mixwithsevenotheranalytes(*),
20μLinjection
Alliance HPLC system with Empower 2 software, PDA detector, separation module with sample manager, column heater, and fluorescence detector.
12
34
5
6
7
8
9 10
11
12
13
14
15
16
17
1819 20
21
22
23
UV at 254 nm
0.16
AU
4
Minutes
6.00 10.00 14.00 18.00 22.00 26.00 30.00 34.00
Figure 3. HPLC analysis of 23 PAHs with UV detection.
peak analyte ppm
1 Napthalene 20
2 Acenaphthylene 40
3 1-Methylnaphthalene* 20
4 2-Methylnaphthalene* 20
5 Acenaphthene 20
6 Fluorene 4
7 Phenanthrene 2
8 Anthracene 2
9 Fluoranthene 4
10 Pyrene 2
11 Benzo(a)anthracene 2
12 Chrysene 2
peak analyte ppm
13 Benzo(e)pyrene* 20
14 Benzo(b)fluoranthene 4
15 Perylene* 20
16 Benzo(k)fluoranthene 2
17 Benzo(a)pyrene 2
18 Dibenzo(a,h)anthracene 4
19 Benzo(g,h,I)perylene 4
20 Indeno(1,2,3-cd)pyrene 2
21 Benzo(b)chrysene* 20
22 Anthanthrene* 20
23 Coronene* 20
Regulated compound in red
*EPA 610 mix with seven other analytes
13
45
6
7
8
9 10
11
12
13
14
15
16
17
18
1920
21
22 23
3000 S
EU
SEU-Sample EnergyUnits
6.00 10.00 14.00 18.00 22.00 26.00 30.00 34.00
Figure 4. HPLC analysis of 23 PAHs with fluorescence detection.
time ex em
Initial 224 330
7.5 270 323
10.0 250 390
12.8 250 420
time ex em
14.8 270 385
16.6 280 410
17.4 385 500
17.8 280 410
time ex em
22.9 305 480
26.0 275 420
29.0 305 480
32.0 300 450
peak analyte ppm
1 Napthalene 2.0
3 1-Methylnaphthalene* 2.0
4 2-Methylnaphthalene* 2.0
5 Acenaphthene 2.0
6 Fluorene 0.4
7 Phenanthrene 0.2
8 Anthracene 0.2
9 Fluoranthene 0.4
10 Pyrene 0.2
11 Benzo(a)anthracene 0.2
12 Chrysene 0.2
13 Benzo(e)pyrene* 2.0
peak analyte ppm
14 Benzo(b)fluoranthene 4.0
15 Perylene* 20.0
16 Benzo(k)fluoranthene 2.0
17 Benzo(a)pyrene 0.2
18 Dibenzo(a,h)anthracene 0.4
19 Benzo(g,h,I)perylene 4.0
20 Indeno(1,2,3-cd)pyrene 2.0
21 Benzo(b)chrysene* 20.0
22 Anthanthrene* 20.0
23 Coronene* 20.0
Regulated compound in red
*EPA 610 mix with seven other analytes
11
0.010
0.020
0.030
226.9
264.7 296.6
0.010
0.020
0.030
226.9
264.7 296.6
-0.00001
0.00000
0.00001
0.00002
Wavelength230.00 250.00 270.00 290.00 310.00
211.7 237.5271.7 312.1
Benzo(a)pyrene
Library match(es)
Difference(s)
Figure 6. Library match triple plot for benzo(a)pyrene.
0.00
10.00
20.00
30.00
40.00
50.00
60.00
70.00
80.00
90.00
20.75 20.85 20.95 21.05 21.15 21.25
AU
Deg
rees
20.75 20.85 20.95 21.05 21.15 21.25
-------
-------
0.002
0.006
0.010
0.014
0.018
0.022
0.026
0.030
0.034
nm220.00 240.00 260.00 280.00 300.00
226.9
264.7
296.6
226.9
264.7296.6
sample spectrum
library spectrum
AU
226.9
264.7
296.6
226.9
264.7296.6
Minutes
PA: 0.166 TH: 3.207Peak: Benzo(a)pyrenePurityNoise
Ben
zo(a
)pyr
ene
- 20.9
40
PurityNoise + Solvent (1.00)
0.009
0.008
0.007
0.006
0.005
0.004
0.003
0.002
0.001
0.000
Purity plot for benzo(a)pyrene PDA identification for benzo(a)pyrene
Figure 5. Purity angle of 0.166 is less than threshold of 3.207.
12
ACQUITY PDA and ACQUITY UPLC FLR detectors with Empower 2 software.
Naphth
ale
ne
- 2.8
66
Ace
naphth
ene
- 4.6
24
Fluore
ne
- 4.8
36
Phen
anth
rene
- 5.7
31
Anth
race
ne
- 5.9
65
Fluora
nth
ene
- 6.5
14
Pyr
ene
- 6.7
63
Chry
sene
- 7.8
00
Ben
zo(a
)anth
race
ne
- 7.8
83
Ben
zo(b
)flu
oranth
ene
- 9.1
65
Ben
zo(k
)flu
ora
nth
ene
- 9.2
90
Ben
zo(a
)pyr
ene
- 9.5
78
Dib
enz(
a,h)a
nth
race
ne
- 10.2
98
Inden
o(1
,2,3
-cd)p
yren
e -
10.7
77
Ben
zo(g
,h,i)p
eryl
ene
- 10.9
38E
U
0.00
200.00
400.00
600.00
800.00
1000.00
1200.00
1400.00
1600.00
1800.00
2000.00
2200.00
2400.00
2600.00
Minutes0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00
Figure 8. PAHs on ACQUITY UPLC are completed up to three times faster than HPLC with acceptable resolution.
3.3.3. Waters acQuity uplc system
a. tuv detector
chromatographic conditions:
Instrument: ACQUITYUPLCwithTUVdetector
Column: ACQUITY™BEHC181.7mm,2.1mmIDx100mm
Mobilephase:Waterandacetonitrilegradient
Flowrate: 400μL/min
Detection: UVmaxplot
Sample: PAHstandardmixtureof10ppmeachanalyte
Injection: 5μLFluorescencedetector
b. fluorescence detector
chromatographic conditions:
Instrument: ACQUITYUPLCsystemwithACQUITYFLR
Sample: PAH610-QC,100:1inacetonitrile
Column: 2.1mmx150mmACQUITYBEHRP181.7μm
Flowrate: 600μL/min
Temperature: 45°C
MobilephaseA:water
MobilephaseB: acetonitrile
peak analyte
1 Napthalene
2 Acenaphthalene
3 1-Methylnaphthalene
4 2-Methylnaphthalene
5 Fluorene
6 Acenapthene
7 Phenanthrene
8 Anthracene
9 Decafluorobiphenyl
10 Fluoranthene
11 Pyrene
peak analyte
12 p-Terphenyl-d14
13 Chrysene
14 Benzo(a)anthracene
15 Decafluorobiphenyl
16 Benzo(b)fluoranthene
17 Benzo(k)fluoranthene
18 Benzo(a)pyrene
19 Dibenzo(a,h)anthracene
20 Indeno(1,2,3-cd)pyrene
21 Benzo(g,h,i)perylen
Regulated compound in red
Minutes2.00 3.00 4.00 5.00 6.00 7.00
1
2
3
4
5
6
7
8
9
1011 12
13
14
15
16
1718
19
20
21
Figure 7. ACQUITY analysis of 21 PAHs with UV detection.
13
4. endoc rine disruptors (edc s)
4.1. introduction of endocrine disruptors
4.1.1. What are endocrine disruptors?
Until the early 1990s, non-polar hazardous compounds, i.e.
persistent organic pollutants (POP) and heavy metals, were the
focusofinterestandawarenessasprioritypollutantsand,conse-
quently,werepartofintensivemonitoringprograms.Today,these
compoundsarelessprevalentfor industrializedcountriessincea
drastic reduction of emissions has been achieved because of the
adoptionofappropriateregulatorymeasuresandtheeliminationof
thedominantpollutionsources.
However, theemissionofso-called“emerging”or“new”unregu-
lated contaminants have emerged as an environmental problem
andthereisawidespreadconsensusthatthiskindofcontamination
mayrequirelegislativeintervention.Thisgroupofcontaminantsis
mainlycomprisedofproductsusedinlargequantitiesineveryday
life,suchashumanandveterinarypharmaceuticals,personalcare
products, surfactants and surfactant residues, plasticizers and
different industrialadditives.Thesecontaminantsdonotneed to
be persistent in the environment to cause negative effects since
their high transformationand removal rates canbe compensated
bytheircontinuousintroductionintoenvironment.Oneofthemain
sourcesofemergingcontaminantsisuntreatedurbanwastewater,
and wastewater treatment plant (WWTP) effluents. Most current
WWTPswerenotdesignedtotreatthesesubstances,andthehigh
portionofemergingcompoundsandtheirmetabolitescanescape
elimination in the WWTP and enter the aquatic environment via
sewageeffluents.
The partial or complete closing of water cycles is an essential
part of sustainable water resources management. The increasing
scarcity of pristine waters for drinking water supply and
increasing use of water by industry and agriculture needs to be
counteredbytheefficientandrationalutilizationofresources.One
optionistorecycletheeffluentsforvariouspurposes,especially
inindustrialandagro/foodproductionactivities.However,dueto
the high costof end-of-pipeapproach (drinkingwater treatment),
thefutureofindirectportablereuserequiresanefficienttreatment
of wastewaters prior to their discharge. Thus, the occurrence of
traceorganic contaminants inwastewaters, theirbehaviorduring
wastewater treatment and drinking water production are the key
issuesthatrequirefurtherstudy.
4.1.2. analysis of emerging contaminants in Wastewaters
Oneofthemajorlimitationsintheanalysisofemergingcontami-
nantsisthelackofanalyticalmethodsforquantificationofthese
contaminantsatlowconcentrations.Theprerequisiteforproperrisk
assessment and monitoring of waste, surface and drinking water
quality is the availability of a multi-residual analytical method
thatpermitsmeasurementat the low (orevenbelow)ng/L level.
However,thefactthatthesecompoundsarenotontheregulatory
lists as environmental pollutants has resulted in comparatively
little attentionbeing received. Consequently, analyticalmethod-
ology for different groups of emerging contaminants is evolving
and the number of methods described in the literature for the
determinationofemergingcontaminantshasgrownconsiderably.
Still, the analysis of this group of contaminants requires further
improvementsintermsofsensitivityandselectivity,especiallyfor
verycomplexmatrices,suchaswastewater.
14
methods for the analysis of acidic pharmaceuticals in wastewaters
compounds extraction derivatizationcHromatoGrapHic
metHoddetection od(ng/l)
Bezafibrate, diclofenac, ibuprofen, gemfibrozil, carbamezapine
Sequential SPE (C18 + polymeric sorbent) - LC MS 2
Salicylic acid, ibuprofen, ketoprofen, naproxen, bezafibrate, diclofenac SPE (polymeric sorbent) - LC MS 5-56
Bezafibrate, clofibric acid, diclofenac, fenoprofen, gemfibrozil, ibuprofen, inomethacin, ketoprofen, naproxen
SPE (C18) - LC MS/MS 5-20
Bezafibrate, clofibric acid, ibuprofen SPE (MCX or polymeric sorbent) - LC MS/MS 0.016-2.18
Ibuprofen, clofibric acid, ketoprofen, naproxen, diclofenac SPE (HLB) diazomethane GC MS 0.3-4.5
Clofibric acid, diclofenac, ibuprofen, phenazone, propyphenazone SPE (C18) Pentaflorobenyl bromide GC MS 0.6-20
Clofibric acid, naproxen, ibuprofen SPE (HLB)BSTFA (bis(trimethylsilyl)-triflouroacetamide)
GC MS 0.4-2.6
Ibuprofen, naproxen, ketoprofen, tolfenamic acid, diclofenac, meclofenamic acid
SPE (HLB)MTBSTFA (N-methyl-N- (tert-butyldimethylsiy) trifluoroacetamide)
GC MS 20
4.1.3. analysis of edcs in drinking Water
Manysyntheticchemicalshavebeensuspectedtohaveendocrine
disrupting properties, and further studies have shown develop-
mental, reproductive and other health problems in wildlife and
laboratory animals. These compounds include pesticides, includ-
inginsecticides,herbicides,fungicidesandnematocides,products
associated with plastics (bisphenol A, phthalates), pharmaceuti-
cals (drugestrogens suchasbirth control pills,DES, cimetidine),
ordinary household products, including breakdown products of
detergentsandassociatedsurfactants,includingnonylphenoland
octylphenol; and industrial chemicals (polychlorinated biphenyls
(PCBs), dioxin and benzo(a)pyrene). The environmental impact of
these compounds is of great concern, particularly in the aquatic
environment,wherefeminizationofmalefishhasbeenseen.Intake
ofsomeofthesecompoundsbyhumansviafoodordrinkingwater
hasbeenthoughttocauseinfertility.
EDCs have become an emerging area of concern. The endocrine
system produces hormones that regulate development, growth,
reproduction andbehavior. Chemicals that have beendetermined
to be estrogenic include synthetic estrogens, steroids, pesti-
cides, phthalates, alkylphenol ethoxylate surfactants, dioxins,
polychlorinated biphenyls (PCBs), andnatural estrogens, such as
phytoestrogens.Besidessyntheticestrogens,otherpharmaceuticals
havealsobecomeaconcernfortheenvironment,suchasibuprofen
andantibiotics,sulfonamides,penicillinsandtetracyclines.
Anenvironmentalendocrineorhormonedisruptormaybedefined
asanexogenousagentthatinterfereswiththesynthesis,secretion,
transport, binding, action, or elimination of natural hormones in
thebodythatareresponsibleforthemaintenanceofhomeostasis,
reproduction,development,and/orbehavior.Forthepurposeofthis
document, the term “endocrine disruptor” will be used synony-
mouslywithhormonedisruptor.Ofimportancehereistheconcept
thatendocrinedisruptorsencompassmorethanjustenvironmental
estrogens,andincludeanyagentthatadverselyaffectsanyaspect
of the entire endocrine system. Endocrine disruptors are usually
eithernaturalproductsorsyntheticchemicalsthatmimic,enhance
(an agonist), or inhibit (an antagonist) the action of hormones.
Undersomecircumstances,theymayactashypertrophic14(stimu-
latory) agents and tumor promoters. Dose, body burden, timing,
anddurationof exposure at critical periods of life are important
considerations forassessingadverseeffectsofanendocrinedis-
ruptor.Effectsmaybereversibleorirreversible,immediate(acute)
orlatentandnotexpressedforaperiodoftime.
15
Theendocrinesystemincludesanumberofcentralnervoussystem
(CNS)-pituitary-targetorganfeedbackpathwaysinvolvedinregu-
latingamultitudeofbodilyfunctionsandmaintaininghomeostasis.
Assuch, therearepotentiallyseveral targetorgansitesatwhich
a given environmental agent could disrupt endocrine function.
Furthermore, because of the complexity of the cellular processes
involved in hormonal communication, any of these loci could be
involved mechanistically in a toxicant’s endocrinerelated effect.
Thus, impairedhormonalcontrolcouldoccurasaconsequenceof
altered hormone: synthesis, storage/release, transport/clearance,
receptorrecognition/binding,orpostreceptorresponses.
regulatory requirements
All European Union (EU) member states have to enforce the EU
directives.CountrieswishingtojointheEUmustcomplywiththeEU
standards.Directive2000/60/ECgivestheframeworkforwaterpoli-
cy.Thisisaverygeneraltext.TheDecision24555/2001/ECamends
thedirectiveandlists theprioritysubstances tobemonitored.This
appliestoallkindofwater.Directive98/83/EUismorespecific to
drinkingwater.
edcs metHod reportinG limit (ppt)
SteroidsAndrostenedione, Estradiol, Ethynylestradiol, Progesterone, Testosterone
LC/MS 1
PAHsAnthracene, Benzo(a)pyrene GC/MS or LC/MS 1
PesticidesAtrazine, Lindane, DDD, DDE, DDT, DEET, Methoxychlor
GC/MS 0.5-1
PlasticizerBisphenol A GC/MS or LC/MS 5
Fire retardantTCEP GC/MS or LC/MS 1
4.2. Waters solution
4.2.1. pharmaceutical compounds in the environment
analysis of surface and groundwater by lc/ms/ms
Hundreds of active substances are being used in human and
veterinary drug formulas. Because of the broad application field
ofpharmaceuticals,theirresiduescanreachtheenvironmentfrom
several pathways. The main pathway is through urinary or fecal
excretion. However, pharmaceutical manufacturing discharges
should also be considered and, even more, direct introduction
occurs when antibiotics are introduced into the aquacultures.
Pharmaceutical compounds are not completely eliminated in the
sewagetreatmentplantsthus,variableamountsreachsurfaceand
groundwatersamples.Inthelastfewyears,theinterestinassess-
ingthepresenceofpharmaceuticalsintheenvironmenthasbeen
growing.Thereisaspecialneedformethodsfortheirrapid,sensi-
tiveandselectiveanalysisofpharmaceuticalsinwater.
LC/MS/MS systems.
16
SPE (Oasis MCX) condition step:
methanol: water pH=3 wash step: water pH=3
elution step: methanol/ammonia=95/5
REGULAR DRUGS TETRACYCLINES
SamplePre-treatment
Extraction
LC separation
MS detection
Adjust pH of 100 mL sample to pH=3
SPE (Oasis HLB) condition step:
methanol + water wash step: water
elution step: methanol
LC on XterraRP Mobile phase: MeOH/H2O +
ammonium accetate
MS/MS detection Micromass Quattro Premier XE
ESI (+) and ESI (-) mode
LC on Symmetry Shield C8 Mobile phase: MeOH/H2O +
ammonium acetate + formic acid
200 mL sample + 5 mL NaEDTA/MClIVaine buffer
Flow diagram of workflow.
analyte recovery (n=5) in % repeatability (n=5) %rds reproducibility (n=10) %rsd lod in ng/l
Acetylsalicylic acid 195 2 87 81
Bezafibrate 80 3 16 18
Bisprolol 87 5 15 9
Carbamazepine 89 2 4 14
Chloramphenicol 82 5 14 13
Chlortetracycline 95 6 22 300
Clofibric acid 78 2 18 13
Dehydro-erythromycine 96 5 29 20
Diclofenac 80 3 6 14
Doxycycline 91 14 41 100
Fenofibrate 36 7 18 22
Ibuprofen 62 9 16 15
Metoprolol 97 2 27 13
Oxytetracycline 97 8 32 100
Paracetamol 83 3 17 45
Sulphamethoxazole 63 12 29 15
Tetracycline 98 6 21 100
Method characteristics.
17
4.2.2. pharmaceutical compounds in the environment
confirmation of residues of lc/Q-tof
Liquid chromatography and quadrupole-orthogonal acceleration
time-of-flight tandem mass spectrometry are used in order to
confirm the proposed identity of pharmaceuticals monitored in
surfaceandgroundwatersamples.Basedontheaccuratemass,the
elementalcompositionsfortheprecursorandproductionscanbe
calculated.Forthefinalconfirmationoftheidentityoftheanalyte,
two product ions (two MS/MS ions) are monitored and the ratios
betweentheionsarecalculatedandcomparedwiththoseofstan-
dardsandfortifiedsamples.Theratiosarecomparablewithinthe
tolerancesoftheEUcriteriacommissiondecision2002/657.
confirmation
LC/Q-Tofisaverypowerfultechniqueforsimultaneousscreening,
identificationandconfirmationof(drug)residues.
ACQUITY UPLC system with the QTof Premiermass spectrometer.
2318
13+14
1516
17 20
1
2
3+45
6
7
8+9
10
11
12
1921
22
Time
%
1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.000
100
UPLC-TOF total ion chromatogram showing the separation of 23 pharmaceutical compounds analyzed in PI mode (100 ng/mL standard solution)
1-Acetaminophen2-Sotalol3-Famotidine4-Atenolol5-Ranitidine6-Trimethoprim7-Ofloxacine8-Sulphametaxozole9-Metoprolol10-Azithromycin11-Propranolol12-Pravastatin
13-Carbamazepine14-Propyphenazone15-Erythromycin16-Bezafibrate17-Ketoprofen18-Paroxetine19-Lansoprazole20-Fluoxetine21-Mefenamicacid22-Loratadine23-Mevastatin
Time0
1007
4
1
6
5
12
10
911
2
3
8
%
1.00 2.00 3.00 4.00 5.00 6.00 7.00
1-Acetaminophen2-Famotidine3-Clofibricacid4-Naproxene5-Ketoprofen6-Bezafibrate7-Diclofenac8-Ibuprofen9-Indomethacin10-Mefenamicacid11-Lansoprazole12-Gemfibrozil
UPLC-TOF total ion chromatogram showing the separation of 12 pharmaceutical compounds analyzed in NI mode (100 ng/mL standard solution).
Time2.00 4.00 6.00 8.00
0
100 1.43
2.00 4.00 6.00 8.000
100
2.00 4.00 6.00 8.000
100
2.00 4.00 6.00 8.000
100 5.84
5.66
5.00
Time2.00 4.00 6.00 8.00
0
1005.78
Carbamezapine
2.00 4.00 6.00 8.000
1003.12
m/z80 100 120 140 160 180 200
%
0
100 93.0344
110.0602
m/z100 200 300 400 500
0
100230.1160
123.0665
261.1346
275.1155
291.1435%
m/z100 200 300 400 500 600 700
%
0
100 591.4233
158.0944 573.5130 749.5177592.4300
750.5252
m/z100 200 300 400 500 600 700
%
0
100 576.3754
158.0938
558.3660 734.4704577.3840
100 150 200 250 300 350m/z
%
0
100 189.1033
231.1504
201.1030
100 150 200 250 300 350m/z
%
0
100 194.1585
192.1551179.0875
237.1036
Time2.00 4.00 6.00 8.00
0
100 1.43
2.00 4.00 6.00 8.000
100
2.00 4.00 6.00 8.000
100
2.00 4.00 6.00 8.000
100 5.84
5.66
5.00
Time2.00 4.00 6.00 8.00
0
1005.78
2.00 4.00 6.00 8.000
1003.12
Time2.00 4.00 6.00 8.00
0
100 1.43
Time2.00 4.00 6.00 8.00
0
100 1.43
2.00 4.00 6.00 8.000
100
2.00 4.00 6.00 8.000
100
2.00 4.00 6.00 8.000
100 5.84
5.665.66
5.00
Time2.00 4.00 6.00 8.00
0
1005.78
2.00 4.00 6.00 8.000
1003.12
2.00 4.00 6.00 8.000
1003.12
m/zTime2.00 4.00 6.00 8.00
0
100 1.43
2.00 4.00 6.00 8.000
100
2.00 4.00 6.00 8.000
100
2.00 4.00 6.00 8.000
100 5.84
5.66
5.00
Time2.00 4.00 6.00 8.00
0
1005.78
2.00 4.00 6.00 8.000
1003.12
m/z80 100 120 140 160 180 200
%
0
100 93.0344
110.0602
m/z100 200 300 400 500
0
100230.1160
123.0665
261.1346
275.1155
291.1435%
m/z100 200 300 400 500 600 700
%
0
100 591.4233
158.0944 573.5130 749.5177592.4300
750.5252
m/z100 200 300 400 500 600 700
%
0
100 576.3754
158.0938
558.3660 734.4704577.3840
100 150 200 250 300 350m/z
%
0
100 189.1033
231.1504
201.1030
100 150 200 250 300 350m/z
%
0
100 194.1585
192.1551179.0875
237.1036
Time2.00 4.00 6.00 8.00
0
100 1.43
2.00 4.00 6.00 8.000
100
2.00 4.00 6.00 8.000
100
2.00 4.00 6.00 8.000
100 5.84
5.66
5.00
Time2.00 4.00 6.00 8.00
0
1005.78
2.00 4.00 6.00 8.000
1003.12
Time2.00 4.00 6.00 8.00
0
100 1.43
Time2.00 4.00 6.00 8.00
%
0
100 1.43
Acetaminophen
2.00 4.00 6.00 8.00
%
0
100
2.00 4.00 6.00 8.00
%
0
100
2.00 4.00 6.00 8.00
%
0
100 5.84
5.665.66
5.00
Azithromycin
Erythromycin
Propyphenazone
Time2.00 4.00 6.00 8.00
%
0
1005.78
2.00 4.00 6.00 8.000
1003.12
2.00 4.00 6.00 8.00
%
0
1003.12
Trimethoprim
m/z
Confirmation of identity of target com-pounds in real wastewater samples.
Leftpanel:Narrowwindow(20mDa)extractedionchromato-gramsof[M+H]obtainedintheTOFmodefor:m/z152.071(acetaminophen)m/z291.146(trimethoprim)m/z749.516(azithromycin)m/z734.468(erythromcyn)m/z231.150(propyphenazonem/z237.103(carbamazepine).
Rightpanel:ProductionspectraobtainedintheQ-TOFmode.
18
4.2.3. a confirmatory method for the determination of
synthetic pyrethroids in Wastewater
n Syntheticpyrethroidswereintroducedtoprovidealesshazard-
ousandlesspersistentalternativetoorganochlorineand
organophosphateinsecticides.
n Theyareamongthesafestinsecticidesavailablebecauseof
theirlowacutetoxicitytomammals.Pyrethroidformulations
areusedinagriculture,animalhusbandry,horticulture,and
inthehome.
n Theyareextensivelyusedformothproofingwoolenmaterials
inthetextileindustryandastheactiveingredientincommer-
cialsheepdips.
eu regulation
some of the synthetic pyrethroids are listed in the dangerous
substances directive (76/464/eec):
list 1.Substanceshavingthepotentialtocausethemost
harmtoaquaticlifeduetotheirpersistence,toxicityor
bioaccumulation.Thedirectiverequirestheeliminationof
dischargesofthesesubstancestotheenvironmentinorder
toreducepollution.
list 2.Substancesthatarethoughttobeharmful,butnotto
thesamedegreeasList1substances.Dischargesofthese
substancesmustbereduced.
cyfluthrin and permethrin are classified under list 2
the discharge of these two compounds into the aquatic environ-
ment should be reduced if:
n Thereisaneedforenvironmentalmonitoring
n EnvironmentalQualityStandards(EQS)cyfluthrin,
0.001mg/L,permethrin,0.01mg/L
n Othersyntheticpyrethroids
n Bifenthrin,l-cyhalothrin,cypermethrin,
fenvalerateanddeltamethrin
sample analysis
1. extraction of wastewater samples
n 10mLhexaneaddedto1Lofsample
n Sampleisshakenfor20minutes
n Theorganiclayerisremoved
n Hexanefractionisthenpassedthroughaconfidentialamino
SPEcleanupprocedure
n SPEcartridgeiswashedwithpentanetoremovenon-polar
matrixinterferences
n Syntheticpyrethroidselutedwithhexane/IPA
n 100timesconcentrationfactorintheextractionmethod
2. chromatographic conditions:
Instrument:MicromassQuattromicro™GC
(tandemquadrupoleGC/MS/MS)
Injector: CryocooledPTVinsolventventmode,10mL,injected
Column: AgilentJ&W,DB-5ms,
30mx0.25mm,IDx0.25m
Flowrate: 1.0mL/minhelium,constant
temperature program:
n 0min100°C
n 2min100°C
n 6min200°Cat25°C/min
n 16min300°Cat10°C/min
Quattro micro GC.
1�
MRM method parameters for EI mode.
MassLynx method window showing how MRM transitions are arranged into five function windows. These functions can be overlapped slightly to allow for small changes in retention time.
Total Ion Chromatogram (TIC) of seven pyrethroids in EI mode.Sensitivity of MRM in EI mode for permethrin and cyflurthrin at a concentration equivalent.
ms method - negative chemical ionization
syntHetic pyretHroid mrm transition dWell time collision enerGy
Bifenthrin 181→166181→165
0.1s0.1s
15eV20eV
λ-Cyhalothrin 197→141181→152
0.1s0.1s
10eV20eV
Permethrin 183→153183→168
0.05s0.05s
12eV12eV
Cyfluthrin/Cypermethrin
163→127163→91
0.05s0.05s
5eV10eV
Fenvalerate 167→125125→89
0.1s0.1s
10eV15eV
Deltamethrin 253→93183→152
0.1s0.1s
15eV20eV
Permethrin≈0.01µg/L10µLinjected
Cyfluthrin≈0.001µg/L10µLinjected
20
5. formaldeHyde analysis
5.1 introduction of formaldehydes
Formaldehyde is an important industrial chemical used to make
other chemicals, building materials, and household products. It
isoneof the large familyof chemical compounds calledvolatile
organic compounds or ‘VOCs’. The term volatile means that the
compounds vaporize, that is, become gaseous, at normal room
temperatures.Formaldehydeservesmanypurposesinproducts.
it is used as a part of:
n Glueoradhesiveinpressedwoodproducts(particleboard,
hardwood,plywood,andmediumdensityfiberboard(MDF)
n Preservativesinsomepaints,coatings,andcosmetics
n Coatingthatprovidespermanentpressqualitytofabrics
anddraperies
n Finishusedtocoatpaperproducts
n Certaininsulationmaterials(urea-formaldehydefoam
andfiberglassinsulation)
Formaldehydeisreleasedintotheairbyburningwood,keroseneor
naturalgas,fromautomobiles,andfromcigarettes.Formaldehyde
canbereleasedfrommaterialsmadewithit.Itisalsoanaturally
occurringsubstance.
Formaldehydeisacolorless,strongsmellinggas.Whenpresentin
theairatlevelsabove0.1ppm(partsinamillionpartsofair),it
cancausewateryeyes,burningsensations in theeyes,noseand
throat, nausea, coughing, chest tightness, wheezing, skin rashes,
andallergicreactions.Ithasalsobeenobservedtocausecancerin
scientificstudiesusinglaboratoryanimals,andmaycausecancer
inhumans.Typicalexposurestohumansaremuchlower;thusany
riskofcausingcancerisbelievedtobesmallatthelevelatwhich
humansareexposed.
regulation limit of each country:
ASHRAE* 0.1ppm
USHUD** 0.4ppm
WHO***Europe 0.08ppm
Sweden 0.1ppm
Japan 0.08ppm
*ASHRAE(AmericanSocietyofHeating,Refrigerating&AirConditioningEngineers)
**HUD(DepartmentofHousingandUrbanDevelopment)
***WHO(WorldHealthOrganization)
analytical method
epa metHod summary
sample matrix field samplinG sample prep Hplc
To-11 Ambient airSep-Pak DNPH-silica cartridge
Backflush cartridge with CH3CN
Two C18 columns 60-100% CH3CN linear gradient
554 Drinking water -DNPH reagent extract
with C18 cartridge
C18 column70-100% CH3OH
linear gradient
8315 Solid waste water -
Same as water or MeCl2
extraction option
Two C18 columns 60-100% CH3CN linear gradient
21
2,4-dinitrophenylhydrazine (dnpH) derivatization method
Waters cartridges for aldehydes and ketones analysis in air
air sampling flow schematic with ozone scrubber cartridge
derivatization reaction (same derivatization used in niosH
method 2532, epa to-11, ip-6a and astm d51�7)
Waters dnpH/xposure™ cartridges air sampling
procedure for aldehydes
n Connectfemaleleur-tippfcartridgetointakeportofasuitable
pumpanddrawairsamplethroughcartridge.
n Fillaleur-tippedsyringewith10mLacetonitrileandconnect
femaleendofcartridgetothesyringetip.
n Pushtheacetonitrilethroughattherateoflessthan
3mL/min.
n Collectinto10mLvolumetricflaskandmakevolume
uptoppingto10mLacetonitrile.
n AnalyzebyHPLC.Air
sample in
Ozone scrubbercartridge
Vacuumsampling
pump
DNPH-Silicaor XPoSurecartridge
Ozone negatively impacts on the analysis of carbonyl compounds in air samples when drawn through cartridges containing silica coated with DNPH. Waters Ozone Scrubber cartridges are designed to remove the ozone interference.
H2 CHNN = NNHH+ R1
R
R1
RNO2
NO2 NO2
NO2 + H2OC = O
DNPH derivative(aldehydeor ketone)
2,4-Dinitrophenylhydrazine(DNPH)
Sep-Pak DNPH cartridge
22
5.2. Waters solution
5.2.1. Hplc analysis
Instrument: BreezeBinaryGradientorAlliance
Column: Nova-Pak®C18,3.9mmx75mm
Mobilephase: Water/acetonitrile/tetrahydrafuran65/35/5/vv
Flowrate: 1.5mL/min
UV/Visdetector: 360nm
chemistry
Sep-PakDNPHcartridge
WatersXPoSurealdehydesamplercartridge
Ozonescrubber
Breeze binary gradient system
0 2 4 6 8 10 12 14Time (minutes)
1
2
3
4
5 6Lab Air Sample
Cartridge Blank
Low level examples aldehyde profile from laboratory airs 100 L sample collected at 0.65 L/min.
lab air
peak analyte ppbv
1 DNPH
2 Formaldehyde-DNPH 4.8
3 Acetaldehyde-DNPH 1.2
4 Acetone-DNPH 118.0
5 Butanone-DNPH 0.8
6 Isovaleraldehyde-DNPH 0.7
blank (calculated for 100l sample)
peak analyte ppbv
2 Formaldehyde-DNPH 0.35
3 Acetaldehyde-DNPH 0.27
4 Acetone-DNPH 0.34
Alliance HPLC system.
23
5.2.2. Waters acQuity uplc method for formaldehyde
chromatographic conditions
Instrument: WatersACQUITYUPLC
Injectionmode: Fullloop
Loopsize: 5μl(5μlinjectionvolume),use15μlneedle
Weakwash: 5%acqueousacetonitrile-800μl
Strongwash: 50%acqueousacetonitrile-500μl
Sampletemp: 25°C
Detection: UVat360nm
allow two minute equilibration between injections
Column: WatersACQUITYUPLCBEHPhenyl2.1x100
mm,1.7μmat35°C
Eluent: A-90:10water-THF(stabilized)*
B-acetonitrile
*Mix900mlwaterand100mlstabilizedtetrahydrofuran(THF),filteranddegas.
Flowrate: 0.5ml/min
reference
“Fast Analysis of Aldehydes and Ketones by ACQUITY UPLC.”
2006,WatersCorporationApplicationNote:720001500EN.
1
1514
1312
11
1098
7
65
4
3
2
0.2
0 A
U
Minutes
4.00 6.00 12.00 16.00 20.00
1-Formaldehyde 9-Isovaleraldehyde
2-Acetaldehyde 10-Pentanal
3-Acetone 11-o-Tolualdehyde
4-Acrolein 12-p-Tolualdehyde
5-Propanal 13-m-Tolulaldehyde
6-Crotonaldehyde 14-Hexanal
7-Butanal 15-2-5Dimethylbenzaldehyde
8-Benzaldehyde
EPA method 8315 A-Opt. 2 analytes 20 ppm as DNPH derivatives.
1
1211
1098
76
5
43
2
0.2
0 A
U
Minutes
4.00 8.00 12.00 16.00 20.00
1-Formaldehyde 7-Pentanal
2-Acetaldehyde 8-Hexanal
3-Propanal 9-Heptanal
4-Crotonaldehyde 10-Octanal
5-Butanal 11-Nonanal
6-Cyclohexanone 12-Decanal
EPA method 554, 8315 A- Opt. 1 analytes 20 ppm as DNPH derivatives.
ACQUITY UPLC with TUV detector.
24
6. dioxins
6.1. introduction of dioxins
dioxin basic chemistry
“Dioxins”referstoagroupofchemicalcompoundsthatsharecer-
tainsimilarchemicalstructuresandbiologicalcharacteristics.
several hundred of these toxic compounds exist and are members
of three closely related families:
n Thechlorinateddibenzo-p-dioxins(CDDs)
n Thechlorinateddibenzofurans(CDFs)
n Certainpolychlorinatedbiphenyls(PCBs)
CDDsandCDFsarenotproducedintentionallyordeliberately,but
are formed as a by-product of chemical processes. These range
fromnaturalprocessessuchasvolcanoeruptionsandforestfires
toman-madeprocessessuchasmanufacturingofchemicals,pesti-
cides,steelandpaints,pulpandpaperbreaching,exhaustemissions
andincineration.PCBsareman-made,butarenolongerproduced
intheUnitedStatesorEurope.
Dioxinsarereleasedintotheairfromcombustionprocessessuchas
commercialormunicipalwasteincinerationandfromburningfuels
likewood,coaloroil.Dioxinscanalsobeformedwhenhousehold
trashisburnedandduringforestfires.Chlorinebleachingofpulp
andpaper,certaintypesofchemicalmanufacturingandprocessing,
and other industrial processes all can create small quantities of
dioxins.Cigarettesmokealsocontainssmallamountsofdioxins.
Over the past decade, worldwide regulatory bodies have been
workingtogetherwithindustrytodramaticallyreducedioxinemis-
sions.Becausedioxinsareextremelypersistentcompounds,levels
ofdioxinsstillexistintheenvironmentfrombothman-madeand
natural sources, and will take years to decline. A large part of
the current exposures to dioxins in the United States are due to
man-made dioxins from releases that occurred in the past, even
decadesago.Evenifallhuman-generateddioxinscouldsomehow
beeliminated,lowlevelsofnaturallyproduceddioxinswillremain.
Countries worldwide are continuing to look for ways to reduce
dioxinlevelsenteringtheenvironmentandtoreducehumanexpo-
suretothem.
Dioxins are a colorless organic compound containing carbon,
hydrogen,oxygenandchlorine.Polychlorinateddioxinsandfurans
(PCDDs & PCDFs) are a group of 210 single compounds, which
typicallyoccurasamixtureofthedifferentcongeners.Seventeen
of these are regarded as being of toxicological concern, and
2,3,7,8-tetrachlordibenzo-p-dioxin(TCDD)isthemosttoxicofthe
dioxin-likesubstances.Itismeasuredinpartspertrillion(ppt).
Therelativetoxicityofthese17compoundsisintherangeofafac-
torof1000.Thetoxicityofallotherdioxinsisexpressedrelativeto
2,3,7,8-TCDDviaToxicEquivalenceFactors(TEFs).2,3,7,8–TCDD
ToxicEquivalents (TEQs)aredeterminedbymultiplying the com-
poundconcentrationsbytheirrespectiveTEFandsummingthem.
Air from combustion processes
DIOXIN FORMATION
Fuel components Industrial waste incineration
25
Dioxins are not soluble in water and are highly soluble in fat.
Thismeans that theybind to sedimentandorganicmatter in the
environmentandareabsorbedintoanimalandhumanfattytissue.
Inaddition,theyarenotbiodegradablesotheyarepersistentand
bio-accumulateinthefoodchain.Oncereleasedintotheenviron-
ment,throughtheairorwater,theyaccumulateinthefattissueof
animalsandhumans.
PCBs,orpolychlorinatedbiphenylsareanothergroupofchemicals.
They are chlorinated aromatic hydrocarbons that are synthesized
bydirectchlorinatingofbiphenyls.TechnicalPCBmixturesarestill
widespread and present today, such as in transformers, building
materials,lubricants,coatings,plasticizersandinks.Someofthe
PCB compounds have toxicological properties that are similar to
dioxinsandarethereforeoftentermed“dioxin-like”PCBs.
First listed in the Second Annual Report on Carcinogens as
“ReasonablyAnticipatedtobeaHumanCarcinogen”–changedto
“KnowntobeaHumanCarcinogen”intheJanuary2001addendum
totheNinthReportonCarcinogens.TherevisedprofilelistingTCDD
as“KnowntobeaHumanCarcinogen”waspublishedasaresultofa
rulingbytheU.S.CourtofAppealsfortheDistrictofColumbiacircuit
dismissingtherequestforaninjunctiontopreventthelistingofTCDD
asa“KnownHumanCarcinogen”intheNinthReport.
regulation
EPAregulates2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD)asahaz-
ardouswasteandtoxicpollutantundertheCleanWaterAct(CWA),the
SafeDrinkingWaterAct(SDWA),theFederalInsecticide,Fungicide,
and Rodenticide Act (FIFRA), the Resource Conservation and
RecoveryAct(RCRA),theComprehensiveEnvironmentalResponse,
CompensationandLiabilityAct(CERCLA),andtheToxicSubstances
ControlAct (TSCA).A reportablequantityof1 lb. (0.454kg)has
beenestablished for TCDDunderCERCLA. Themaximumcontami-
nantlevelforthechemicalindrinkingwateris3*10-8mg/L.
The U.S. Food and Drug Administration (FDA) regulates TCDD in
bottledwater;theallowableconcentrationisalso3*10-8mg/L.
NIOSHhasrecommendedthattheexposurelimitofTCDDbethelow-
est feasible concentration. OSHA regulates TCDD under the Hazard
CommunicationStandardandasahazardouschemicalinlaboratories.
O
O
OCl
Cl Cl
Cl
2,3,7,8-Tetrachlorodibenzo-p-dioxin
3,3',4,4',5,5'-Hexachlorobiphenyl
2,3,7,8-Tetrachlorodibenzofuran
Dioxin structure
Furan structure
Biphenyl structure
26
6.2. u.s. epa method 1613
tetra- through octa-chlorinated dioxins and furans by isotope
dilution HrGc/Hrms
EPAmethod1613wasdevelopedbytheUSEPA’sOfficeofScience
and Technology for isomerspecific determination of the 2,3,7,8-
substituted,tetra-throughocta-chlorinated,dibenzo-p-dioxinsand
dibenzofurans in aqueous, solid, and tissue matrices by isotope
dilution, high resolution capillary column gas chromatography
(HRGC)/highresolutionmassspectrometry(HRMS).
extraction
summary of method
n Aqueoussamples(samplescontaininglessthan1%solids)
Stableisotopicallylabeledanalogsof15ofthe2,3,7,8-
substitutedCDDs/CDFsarespikedintoa1Lsample,andthe
sampleisextractedbyoneofthreeprocedures:
Samplescontainingnovisibleparticlesareextractedwith
methylenechlorideinaseparatoryfunnelorbythesolid-
phaseextractiontechniqueasdescribedinSection1.1.3.
Theextractisconcentratedforcleanup.
Samplescontainingvisibleparticlesarevacuumfiltered
throughaglassfiberfilter.Thefilterisextractedina
Soxhlet/Dean-Stark(SDS)extractor(Reference7),andthe
filtrateisextractedwithmethylenechlorideinasepara-
toryfunnel.Themethylenechlorideextractisconcentrated
andcombinedwiththeSDSextractpriortocleanup.
Thesampleisvacuumfilteredthroughaglassfiberfilter
ontopofasolidphaseextraction(SPE)disk.Thefilterand
diskareextractedinanSDSextractor,andtheextractis
concentratedforcleanup.
n Solid,semi-solid,andmulti-phasesamples(butnottissue)
–Thelabeledcompoundsarespikedintoasamplecontaining
10g(dryweight)ofsolids.Samplescontainingmultiplephas-
esarepressurefilteredandanyaqueousliquidisdiscarded.
Coarsesolidsaregroundorhomogenized.Anynon-aqueous
liquidfrommulti-phasesamplesiscombinedwiththesolids
andextractedinanSDSextractor.Theextractisconcentrated
forcleanup.
n Fishandothertissue–Thesampleisextractedbyoneof
twoprocedures:
SoxhletorSDSextraction–A20galiquotofsampleis
homogenized,anda10galiquotisspikedwiththelabeled
compounds.Thesampleismixedwithsodiumsulfate,
allowedtodryfor12-24hours,andextractedfor18-24
hoursusingmethylenechloride:hexane(1:1)inaSoxhlet
extractor.Theextractisevaporatedtodryness,andthe
lipidcontentisdetermined.
HCldigestion–A20galiquotishomogenized,anda10
galiquotisplacedinabottleandspikedwiththelabeled
compounds.Afterequilibration,200mLofhydrochloric
acidand200mLofmethylenechloride:hexane(1:1)are
added,andthebottleisagitatedfor12-24hours.The
extractisevaporatedtodryness,andthelipidcontentis
determined.
after extraction
Cl-labeled 2,3,7,8-TCDD is added to each extract to measure
the374efficiencyof the cleanup process. Sample cleanupsmay
includeback-extractionwithacidand/orbase,andgelpermeation,
alumina, silica gel, Florisil and activated carbo chromatography.
High-performance liquid chromatography (HPLC) can be used for
further isolation of the 2,3,7,8-isomers or other specific isomers
orcongeners.Prior to thecleanupprocedurescitedabove, tissue
extracts should be cleaned up using an anthropogenic isolation
column,abatchsilicageladsorption,orsulfuricacidandbaseback-
extraction,dependingonthetissueextractionprocedureused.
27
after cleanup
The extract is concentrated tonear dryness. Immediately prior to
injection,internalstandardsareaddedtoeachextract,andanaliquot
oftheextractisinjectedintothegaschromatograph.Theanalytes
are separated by the GC and detected by a high resolution mass
spectrometer.Twoexactm/z’saremonitoredforeachanalyte.
identification
An individual CDD/CDF is identified by comparing the GC reten-
tion time and ion abundance ratio of two exact m/z’s with the
corresponding retention time of an authentic standard and the
theoreticaloracquiredion-abundanceratioofthetwoexactm/z’s.
The non-2,3,7,8 substituted isomers and congeners are identi-
fiedwhen retention timesand ion-abundance ratiosagreewithin
predefinedlimits.Isomerspecificityfor2,3,7,8-TCDDand2,3,7,8-
TCDFisachievedusingGCcolumnsthatresolvetheseisomersfrom
theothertetra-isomers.
Quantitation
Quantitativeanalysisisperformedusingselectedioncurrentpro-
file(SICP)areas,inoneofthreeways:
n Forthe152,3,7,8-substitutedCDDs/CDFswithlabeled
analogs,theGC/MSsystemiscalibrated,andtheconcentra-
tionofeachcompoundisdeterminedusingtheisotope
dilutiontechnique.
n For1,2,3,7,8,9-HxCDD,OCDF,andthelabeledcompounds,
theGC/MSsystemiscalibratedandtheconcentrationofeach
compoundisdeterminedusingtheinternalstandardtechnique.
n Fornon-2,3,7,8-substitutedisomersandforallisomersata
givenlevelofchlorination(i.e.totalTCDD),concentrations
aredeterminedusingresponsefactorsfromcalibrationofthe
CDDs/CDFsatthesamelevelofchlorination.
Qualitation
Thequalityoftheanalysisisassuredthroughreproduciblecalibra-
tionandtestingoftheextraction,cleanupandGC/MSsystems.
6.3. Waters solution
The Micromass AutoSpec Premier™ is the market-leading instru-
ment for High Resolution Gas Chromatography (HRGC) coupled
withHighResolutionMassSpectrometry(HRMS).Itoffersultimate
sensitivity,quantitativelinearityandreproducibilitynecessaryfor
regulatorydioxinandfuranmonitoring.TheAutoSpecPremierpro-
videsunmatchedperformance for theanalysesofdioxins, furans,
polychlorinatedbiphenyls(PCBs),polybrominateddiphenylethers
(PDBEsfrombrominatedflameretardants)andrelatedcompounds.
Theuseofascreeningmethodpriortoconfirmatoryanalysiscan
greatlyreducetheworkloadofaHRGC/HRMSlaboratorybyhighlight-
ingsamplesthatareeithernon-detectable,orhaveconcentrations
atextremelyhighlevelsthatmayfalloutsidethequantifiablerange
oftheHRGC/HRMSmethod.Arapidscreeningmethodforallnon-
toxicandtoxicPCDD/Fsinasingleinjectionhasbeendevelopedby
GCTripleQuadrupoleMS/MS.Wecurrentlyrecommendthat10%of
screenedsamplesareverifiedbyHRGC/HRMS.
Quattro micro GC.
28
6.3.1. dioxin screening using Gc/ms/ms (Quattro micro Gc)
QuattromicroGCisatandemquadrupoleinstrumentforsuperior
quantitative performance. It provides the exceptional sensitivity,
selectivity,androbustnessoftandemquadrupoleGC/MS/MS.The
instrument produces accurate and reproducible quantification of
targetcompoundsatlowlevelsincomplexmatricessuchasfruits,
vegetables and animal products. It provides complete range of
automatedreal-timeMS/MSexperiments:
MultipleReactionMonitoring(MRM),production,precursorionand
neutralloss.QuattromicroGCconsistsofnewionopticanddedi-
catedEIandCIsourceswithisolationvalveforrapidchangeover.
Quattro micro Gc conditions:
GCcolumn:
20m0.18mmI.D.0.18mmdfAgilentJ&W,DB-5ms
0.6mL/minHeflow
GCtemperatureramp:
140°Cfor1.5mins
18.4°C/minto220°C
3.7°C/minto255°C
10.4°C/minto310°Chold1min
Splitlessinjector,2mmIDdeactivatedquartzlinerinjector
temperature280°C
1.6minpurgetime30mL/minpurgeflow
1mLinjectionvolume
QUADRUPOLE(MS1)
HEXAPOLECOLLISION CELL
QUADRUPOLE(MS2)
PHOTOMULTIPLIER
PHOSPHOR
PRE-FILTERPOST-FILTER
CONVERSIONDYNODE
Detector
REMOVEABLEION SOURCE
GC INTERFACE
GC OVEN
Schematic diagram of the Quattro micro GC showing the key components of the tandem quadrupole mass spectrometer.
Mid point calibration standard, TIC and extracted mass chro-matograms for TCDF, 13C12-TCDF.
Mid point calibration standard, TCDF transition 306>243 and 13C12-TCDF transition 318>254.
Separation of 1,2,3,4,7,8-HxCDF and 1,2,3,6,7,8-HxCDF showing <25% valley as required by EU legislation.
2�
temperature ramp optimized to satisfy eu legislation:
n Separationof1,2,3,4,7,8-HxCDFand1,2,3,6,7,8-HxCDFmust
be<25%valley.A13%valleyseparationisobtainedunder
theseconditions.Thischromatogramshowsinjectionnumber
80fromthebatchruns.
n OCDFelutesinlessthan20minutes
n Rapidanalysistime,maintainingrequiredseparation,maxi-
mumthroughput
n Midpointcalibrationstandard,TICandextractedmasschro-
matogramsforTCDF,13C12-TCDF
n Midpointcalibrationstandard,TCDFtransition306>243and
13C12-TCDFtransition318>254
Quattro micro Gc reproducibility
conGener prf %rsd ion ratio %rsd rt %rsd
2378-TCDF 6.35 5.9 0.14
12378-PeCDF 6.39 3.94 0.16
23478-PeCDF 6.81 4.53 0.13
123478-HxCDF 5.34 4.52 0.14
123678-HxCDF 4.78 4.23 0.15
234678-HxCDF 5.21 3.68 0.14
123789-HxCDF 5.06 3.75 0.13
1234678-HpCDF 4.88 5.27 0.11
1234789HpCDF 4.07 4.01 0.09
OCDF 3.48 2.96 0.09
2378-TCDD 6.51 4.99 0.13
12378-PeCDD 5.85 3.75 0.14
123478-HxCDD 4.13 4.36 0.14
123678-HxCDD 6.16 3.92 0.14
123789-HxCDD 4.09 3.53 0.14
1234678-HpCDD 3.6 3.46 0.08
OCDD 3.53 2.24 0.09
6.3.2. dioxin confirmation using Hrms (autospec premier)
TheAutoSpecPremier isbasedonWatersMicromassestablished
AutoSpectechnology.Withan installedbaseofover650instru-
ments, the AutoSpec family features patented tri-sector EBE
technology,ultrahighresolution(>80,000),aconstantgain,long
life detector, plug-in sources, unrivalled dynamic range >105),
advancedautotuneanduncompromisingsensitivity.
next generation informatics for complete dioxin confidence
The AutoSpec Premier is powered by Waters MassLynx software.
MassLynx automates instrument tuning, control and data acquisi-
tionintoasingleplatform. Inaddition,ourTargetLynx™Application
Manager, designed in collaboration with the world’s leading GC/MS
laboratories,providesanintegrateddataprocessingenvironmentfor
dioxin,environmentalandotherrelatedGC/MSquantitativeanalyses.
In addition, our TargetLynx Application Manager, designed in col-
laborationwith theworlds leadingGC/MS laboratories, provides an
integrated data processing and reporting environment for dioxin,
environmentalandotherrelatedGC/MSquantitativeanalyses.
TheTargetLynxapplicationnowsupportsreportingofresultscompliant
withUS EPAmethod1613 reporting protocols. TheReporting tools
allowtheusertoinputawiderangeoftraceabilityparametersdirectly
intoa fully customisable report,withparameters suchas labname,
samplereceiptdate,sampleextractionmethodandmanymore.During
thereportgeneration,itispossiblefortheusertoaddcommentsto
any of the fields on the report,with all comments appended to the
relevant‘table’ofthereport.
Afterinput,alloftheseparametersarethenstoredintheTargetLynx
dataset,maintaininggoodtraceabilityandallowingregenerationofa
reportwithease.
AllUSEPA1613reporttemplatesaresuppliedasstandard,andcanbe
modifiedtogenerateanyreportrequired.
AutoSpec Premier.
30
7. Wat ers sof t Ware solut ions
7.1 masslynx related ms products
masslynxistheheartofthedatasystem.Thefollowingapplication
managersaresuitableforenvironmentalapplications.QuanLynx™
istheMassLynxApplicationManagerdedicatedtohighperformance
quantification. QuanLynx includes the QuanOptimize™ software
tool.AttheheartofQuanLynx,linkingtheextracteddata,calibra-
tions,chromatogramsandresultsisthesummarybar.Thesummary
barshowsallofyourinformationinapowerful,userconfigurable,
spreadsheet-like grid. Using the summary bar, you can navigate
through your data and rapidly check peaks areas, concentrations
andresidualsandmore.Fromthetoolbar,youcanenablethepop-
upstatisticswindowwhereyoucanviewbatchstatisticsincluding
mean,standarddeviationand%RSDofyourcompounds.Thecom-
pleteexperimentalrecordisalsopartoftheQuanLynxcohesivefile
architecture,whichincludesarecordofMSandsampleinformation
importedfromthemethodfilesandsamplelistwhentherawdata
wasfirstexportedintoQuanLynx.
Quanoptimize(includedwiththeQuanLynxoption)generatesMS
andMS/MSparametersbyoptimizingtheconevoltage,parention
and collision energy parameters. QuanOptimize then takes these
MS methods and performs automated acquisition and processing
usingprocessingmethodsdevelopedonthefly.
targetlynx application manager is designed for quantification
withcomprehensivetargetingandconfirmation.TargetLynxincludes
theQuanLynxandQuanOptimizetools.Forregulatoryfoodsafety
monitoring,evidenceisrequiredbeforethepresenceandconcen-
trationofananalytemaybereportedconfirmatory.Retentiontime,
S/Nratioandsecondaryionratioshavetobeverified.TargetLynx
is designed to automatically fulfill the requirements of the EU
Council Directive 96/23/EC and subsequent SANCO regulations.
TargetLynxgeneratesareport,whichclearlyhighlightsthepositive
samples,andflagstheQCparameterswhenoutoflimits.
Wheninjectingaseriesofsamples,theconcentrationsarereported
asfollows:
n Theconcentrationsthatarebelowthereportinglimit(user
defined,i.e.LOQ)arenotreported.
n Theconcentrationsbetweenthereportinglimitandthemaxi-
mumallowedconcentration(MRL)arereported.
n TheconcentrationsabovetheMRLarereportedinbold.
n IftheQCparametersareoutoflimits,theyareflaggedinred,
andtheexplanationappearsasatooltip.
Right click to add comments.
US EPA 1DFA samplesummary report.
Cal curve summary: five injections, 33 compounds, one page.
31
chromalynx™ApplicationManagerisdesignedfortheautomated
processingofGC/MSorGC/MS/MSdata.Thehighchromatographic
separation efficiency and narrow chromatographic peak widths
typicalofGC/MSoftenincreasethechallengetoidentifyco-eluting
components.ThekeytotheexceptionalperformanceofChromaLynx
is thenew,proprietarychromatographicdeconvolutionalgorithm.
Thisprovidesexceptionalefficiencyforthelocation,peakdetection
andgenerationof ‘clean’massspectraof closelyelutingcompo-
nentsincomplexmixtures.Auniquefeatureofthealgorithmisits
abilitytoretainexactmassmeasurements (whenusedwithexact
mass instruments such as the Waters Micromass GCT), enabling
calculationofelementalcompositionfromthe‘clean’deconvoluted
massspectra.Dataisthenavailableforeasyreviewinthededicated
ChromaLynxdeconvolutionbrowser.Deconvolutedmassspectracan
beautomatically librarysearchedagainstcommerciallyavailable
libraries. Using instruments providing exact mass measurement,
theresultingelementalcompositioncalculationsfromexactmass
spectracangiveconfirmationofthelibrarysearchresults.
7.2 empower software
Empower2softwareisWaters’chromatographydatasoftware(CDS)
packageforadvanceddataacquisition,management,processing,and
reporting.Empower2capabilitiesincludecustomizabledatareports,
integrated custom calculations, and the ability to control Waters
UPLC, HPLC, and LC/MS systems, as well as third-party HPLC and
GCinstruments.CombinedwithEmpower2’sintuitiveuserinterface,
thesepowerfulfeatureshelpmaximizelaboratoryefficiency.Options
forEmpower2includeEmpower2MethodValidationManager(MVM)
andEmpower2EnterpriseDataManager(EDM).
Empower 2 MVM allows you to perform chromatographic method
validation, fromprotocolplanning through final reporting,within
oneapplication.Capabilitiesincludestorageofyourmethodvali-
dation SOP requirements in Empower 2 so that Empower 2 can
automaticallyguideyouthroughthemethodvalidationworkflow,
calculatemethodvalidationresults,determineifthedataadheres
toSOPrequirementsandiftheresultsarewithinspecification,and
generatecustomreports–allwhilemanagingthedatainasecure,
audittrailed,compliantenvironment.
Empower 2 EDM is an enterprise option designed for networked
environmentsthatoffersenhanceddatamanagementcapabilities
forchromatographicresults. Itprovidesautomatedprojectarchive
capabilitieswith advancedarchive logic. Fully configurable, EDM
scales from a single lab to the global enterprise without major
modifications toexisting IT infrastructures. Empower2EDMpro-
videsaccesstoCDSreportsanddataviaawebbrowser–enabling
QualityAssurance staff to review and approve electronic reports
withoutadditionalsoftware.
8. Wat ers Global serv ic es
WatersGlobalServicesoptimizesyourentirelaboratoryoperation
andinformationmanagementprocess.Wehelpyoutoachievenew
levelsofefficiencyandproductivitybyintegratingourbreadthof
knowledgein instrumentation,training,support,GxPcompliance,
andinformatics.
With94officesinmorethan50countries,ourlocalizedcustomer
service teams offer highly skilled and experienced professionals
whose mission is to provide you with the level of expertise and
responsivenessthatyouneed.
Ourportfolioincludes:
n Assetmanagement
n Relocationservices
n Educationandtraining
n GxPcomplianceservices
n Instrumentandsoftwareservices
n Enterprisesolutions
n Onlinesupportcenter
n WatersQualityParts®
Waters corporation 34 Maple Street Milford, MA 01757 U.S.A. T: 1 508 478 2000 F: 1 508 872 1990 www.waters.com
Waters,ACQUITYUPLC,Alliance,Sep-Pak,Micromass,Nova-Pak,Symmetry,ConnectionsINSIGHT,andBreezeareregisteredtrademarksofWatersCorporation.AutoSpec,AutoSpecPremier,ChromaLynx,Empower,Envirogel,GCT,MassLynx,Micromass,Q-Tof,Q-TofPremier,QuanLynx,QuanOptimize,QuattromicroGC,QuattroPremier,Sep-Pak,TargetLynx,XpoSure,andZQaretrademarksofWatersCorporation.Allothertrademarksarethepropertyoftheirrespectiveowners.
©2007WatersCorporation.PrintedintheU.S.A.June2007720002163ENJH-AC
SAleS OFFiceS:
austria 43 1 877 18 07
australia 61 2 9933 1777
belgium and luxembourg 32 2 726 1000
brazil 55 11 4134 3788
canada 1 800 252 4752
china 0086 10 8586 8899
czech republic 420 2 617 11384
denmark 45 46 59 8080
finland 358 9 5659 6288
france 33 1 30 48 72 00
Germany 49 6196 400 7000
Hong kong 852 2964 1800
Hungary 36 1 350 5086
india 91 80 2837 1900
ireland 353 1 448 1500
italy 39 02 265 0983
Japan 81 3 3471 7191
korea 82 2 820 2700
mexico 52 55 52 00 1860
the netherlands 31 76 508 7200
norway 47 6 384 6050
poland 48 22 833 4400
puerto rico 787 747 8445
russia/cis 495 727 4490 / 290 9737
singapore 65 6273 1221
spain 34 93 600 9300
sweden 46 8 555 115 00
switzerland 41 21 983 1818
taiwan 886 2 2543 1898
uk 44 208 238 6100
us 1 800 252 4752
