ANALYSIS OF CYTOSTATIC COMPOUNDS · 1 ANALYSIS OF CYTOSTATIC COMPOUNDS IN AQUATIC ENVIRONMENT...
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ANALYSIS OF CYTOSTATIC COMPOUNDS IN AQUATIC ENVIRONMENT Ľubomíra Kovaľová 1 under the supervision of Juliane Hollender 2 and Wolfgang Dott 1 1 RWTH Aachen University, Institute of Hygiene and Environmental Health 2 EAWAG, Swiss Federal Institute of Aquatic Science and Technology 18 January 2006 AQUAbase Workshop on ANALYTICAL METHODS
Transcript of ANALYSIS OF CYTOSTATIC COMPOUNDS · 1 ANALYSIS OF CYTOSTATIC COMPOUNDS IN AQUATIC ENVIRONMENT...
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ANALYSIS OF CYTOSTATIC COMPOUNDSIN AQUATIC ENVIRONMENT
Ľubomíra Kovaľová1
under the supervision ofJuliane Hollender2 and Wolfgang Dott1
1 RWTH Aachen University, Institute of Hygiene and Environmental Health2 EAWAG, Swiss Federal Institute of Aquatic Science and Technology
18 January 2006 AQUAbase Workshop on ANALYTICAL METHODS
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• CONTEXT• COMPOUNDS
• ANALYTICAL CHALLENGE
• ANALYTICAL METHOD
• OUTLOOK
PHARMACEUTICALS IN THE ENVIRONMENT
- GENERAL AIM:
- find out ENVIRONMENTAL CONCENTRATIONS
- check ECOTOXICOLOGY
- provide ways of ELIMINATION FROM ENVIRONMENT (if necessary)
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• CONTEXT
• COMPOUNDS• ANALYTICAL
CHALLENGE
• ANALYTICAL METHOD
• OUTLOOK
CYTOSTATICS
- COMPOUNDS USED IN CHEMOTHERAPY OF PATIENTS WITH CANCER
- DESIGNED TO CAUSE MUTATION AND KILL CELLS
- PRESCRIPTION RATE:
Denmark 2001 cytostatic kg/year
HYDROXYCARBAMID 180
5-FLUOROURACIL 8.4
CYCLOPHASPHAMIDE 6.8
METHOTREXATE 1.0
IFOSFAMIDE 0.9
ETOPOSIDE 0.7
Uniklinik Aachen 2004cytostatic kg/year
5-FLUOROURACIL 2.9
CYTARABINE 1.0
IFOSFAMIDE 1.0
CYCLOPHOSPHAMIDE 0.8
GEMCITABINE 0.7
MEHOTREXATE 0.6
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• CONTEXT
• COMPOUNDS• ANALYTICAL
CHALLENGE
• ANALYTICAL METHOD
• OUTLOOK Uniklinik Aachen 2004cytostatic kg/year
5-FLUOROURACIL 2.9
CYTARABINE 1.0
GEMCITABINE 0.7
HOSPITAL WASTEWATERµg/l
7.4
2.7
1.8
N
N
O
O
N
O
O
O
N
N
O
O
F
N
O
O
F
N
N
O
O
F
5-FLUOROURACIL CYTARABINE GEMCITABINE
unmetabolised ng/l
740
270
180
10%
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ACIDIC ACIDIC ACIDIC ACIDIC
ACIDIC BASIC BASIC
• CONTEXT
• COMPOUNDS• ANALYTICAL
CHALLENGE
• ANALYTICAL METHOD
• OUTLOOK
N
N
O
O
F
N
N
O
O
N
O
O
O
N
N
O
O
F
N
O
O
F
5-FLUOROURACIL CYTARABINE GEMCITABINE
Metabolites:
5-fluorouridine 5-fluoro- uracil 1-β-D- 2’,2’-difluoro-2′-deoxyuridine arabinofuranoside deoxyuridine
N
N
O
O
O
O
O
O
F
N
N
O
O
O
O
O
F
N
N
O
O
O
O
O
O
N
N
O
O
O
O
O
F
F
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• CONTEXT
• COMPOUNDS
• ANALYTICAL CHALLENGE
• ANALYTICAL METHOD
• OUTLOOK
ANALYTICAL CHALLENGE
- VERY POLAR COMPOUNDS
Log Kow -0.8 to -2.3
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• CONTEXT
• COMPOUNDS
• ANALYTICAL CHALLENGE
• ANALYTICAL METHOD
• OUTLOOK
ANALYTICAL METHOD
CLEAN-UP/PRECONCEN- SEPARATION DETECTION
TRATION
SPE HPLC MS/MS
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• CONTEXT
• COMPOUNDS
• ANALYTICAL CHALLENGE
• ANALYTICAL METHOD
• OUTLOOK
ANALYTICAL METHOD
HPLC SEPARATION I.RP columns separation retention
Synergi HydroPhenomenex
AquaPhenomenex
Polaris C18-EtherVarian
Synergi PolarPhenomenex
Nucleosil C18Machery-Nagel
MZ - PAHMZ
Symmetry C18Waters
RP – Reversed Phase
CLE
AN
-UP
DET
ECTI
ON
mAU
0
10
20
30
40
50
60
0 2 4 6 min
CytR GemC
5-Fu
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• CONTEXT
• COMPOUNDS
• ANALYTICAL CHALLENGE
• ANALYTICAL METHOD
• OUTLOOK
ANALYTICAL METHOD
HPLC SEPARATION II.
HILIC columns separation retention
Zorbax NH2Agilent
TSK-Gel Amide 80TOSOH Bioscience
ZIC-HILICSeQuant
HILIC – Hydrophilic Interaction Chromatography
CLE
AN
-UP
DET
ECTI
ON
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• CONTEXT
• COMPOUNDS
• ANALYTICAL CHALLENGE
• ANALYTICAL METHOD
• OUTLOOK
ANALYTICAL METHOD
DETECTION
5-Fu CytR GemC
- MS/MS (electrospray ionisation)
- UV
CLE
AN
-UP
SEPA
RA
TIO
N
LOQ [µg/l]
TSQ Quantum 50 5 10
API 4000 0.1 0.5 0.5
HP 1100 300 500 500
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• CONTEXT
• COMPOUNDS
• ANALYTICAL CHALLENGE
• ANALYTICAL METHOD
• OUTLOOK
ANALYTICAL METHOD
CLEAN-UP / PRECONCENTRATION
RP mechanism (pH adjusted to keep analytes neutral)
acidic basicanalytes analytes
ISOLUTE ENV+IST
Strata-XPhenomenex
Oasis HLBWaters
SEPA
RA
TIO
N
DET
ECTI
ON
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• CONTEXT
• COMPOUNDS
• ANALYTICAL CHALLENGE
• ANALYTICAL METHOD
• OUTLOOK
ANALYTICAL METHOD
CLEAN-UP / PRECONCENTRATION
Cation Exchange mechanism (pH adjusted to keep analytes positively charged)
basic analytesBond Elut CertifyVarian
Accubond EvidexAgilent
Bakerbond Narc-2JT Baker
Strata-XCPhenomenex
Empore MPC-SD3M
SEPA
RA
TIO
N
DET
ECTI
ON
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• CONTEXT
• COMPOUNDS
• ANALYTICAL CHALLENGE
• ANALYTICAL METHOD
• OUTLOOK
ANALYTICAL METHOD
CLEAN-UP / PRECONCENTRATION
Cation Exchange mechanism (pH adjusted to keep analytes positively charged)
basic analytesSPEC MP1Ansys
SPEC MP3Ansys
Speedisk H2O-philic SC-DVBJT Baker
SEPA
RA
TIO
N
DET
ECTI
ON
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• CONTEXT
• COMPOUNDS
• ANALYTICAL CHALLENGE
• ANALYTICAL METHOD
• OUTLOOK OUTLOOK
- to complete analytical method:choice of SPE materialsoptimization of SPE procedurechoice of internal standards
SUMMARY
- good HPLC separation and retention using HILIC- satisfactory detection - selection of SPE retention mechanism for basic compounds
ENVIRONMENTAL CONCENTRATIONS
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• CONTEXT
• COMPOUNDS
• ANALYTICAL CHALLENGE
• ANALYTICAL METHOD
• OUTLOOK
thank you for your attention
