Photodegradation, Aerobic Biodegradability and
Identification of Transformation Products of
Thalidomide by LC–MS/MS
Waleed M. M. Mahmoud1,2,Christoph Trautwein3, Klaus Kümmerer1,*
1.
Institute of Sustainable and Environmental Chemistry, Faculty of Sustainability, Leuphana University Lüneburg, Scharnhorststraße 1/C13, DE-21335 Lüneburg, Germany
2. Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Suez Canal University, Ismailia 41522, Egypt
3. Department of Environmental Health Sciences, University Medical Center Freiburg, Breisacher Strasse 115B, D-79106 Freiburg, Germany
Introduction
Pharmaceutical compounds are released into the environment in a variety of ways: via wastewater effluent as a result of incomplete metabolism in the body after use in human
therapy, through improper disposal by private households or hospitals or through insufficient removal by sewage treatment plants (Gomez et al., 2007; Gros et al., 2006).
Thalidomide (TD), the drug notorious for its teratogenicity which can be developed after a single dose, has immunomodulating and anti-inflammatory activity. In 1998, TD was
approved by the FDA for the treatment of erythema nodusum leprosum associated with leprosy. TD has many adverse effects. (Sweetman, 2009).
TD is sensitive to hydrolytic decomposition, its hydrolysis products which contain the intact phthalimide moiety showed teratogenic activity (Reist et al., 1998). Hence, it is very
important to monitor the behavior of TD and its degradation or metabolic products in the aquatic environment.
Objective
Microorganisms
UV- light
N
O
NH
O
• TD was not readily biodegraded in Closed bottle test (CBT, OECD 301D) and Manometric
respiratory test (MRT, OECD 301F). But it has a partial degree of degradation in both CBT and
MRT leading to formation of new biodegradation products.
• TD photodegradation lead to intermediary compounds, although presents a partial degree of
mineralization, so it can be an important loss process in limiting TD persistence in the aquatic
environment. Toxicological studies will be conducted for TD and its photodegradation products in
order to perform a risk assessment.
micro
O
TD
Conclusion
The objective of this study is to assess the
stability of TD during photoirradiation (Fig. 1, 2)
and ready biodegradability tests (Fig 3, 4 ) and
to correlate this information for use in predicting
its environmental behaviour and consequences
thereof.
O
???
Materials and Methods
Table 1. Tests for Biodegradability
OECD
Compound
Bacterial
(biodegradability) Concentration density
Test
MRT
OECD
301F
“ready”
surface water
Increase
“ready”
Increase
CBT
OECD
301D
“Simulated”
environmental
compartment
sewage treatment
plant effluent, and
aerobic sediment
Bottles from CBT (Clear),
and MRT (Brown)
Primary elimination of TD was monitored
and structures of photo-products were
assessed by Shimadzu LC-UV at 220nm
and HPLC 1100 Agilent connected to
Bruker Daltonic Esquire 6000 plus ion-trap
mass spectrometer (IT-MS).
Photodegradation tests for TD
were performed with mediumpressure mercury lamp (TQ150,
UV Consulting Peschl).
100,00
100
90
80
70
60
50
40
30
20
10
0
DOC
LC-UV 220nm
LC-FL 225ex,398em
80,00
Degradation (%)
%Recovery
Results
60,00
40,00
20,00
0,00
0
7
14
21
28
-20,00
Time (days)
0
2
4
8
16
32
64
128
Thalidomide
Mercury lamp exposure time (min)
Fig.1. Time course of recovery % of TD concentration by DOC, LC-UV at 220nm and
LC-FL (EX 225, EM 398) during photodegradation. TD conc. 10mg L-1 (n=2).
Quality control
Toxicity Control Calculated
Toxicity Control measured
Fig.3. Closed bottle test (CBT) of Thalidomide. (n=3)
120
Degradation (%)
100
80
60
40
20
0
0
2
4
6
8
10
12
14
16
18
20
22
24
26
28
-20
Time (days)
Fig.2. a) Extracted ion chromatogram of mass 259.1 m/z. B) MS2 of PP1, PP2
and TD. PP (Photodegradation product).
Thalidomide
Measured toxicity control
Fig.4. Manometric respiratory test (MRT) of Thalidomide.
References
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Acknowledgments
Gomez, M.J., Martinez Bueno, M.J., Lacorte, S., Fernandez-Alba, A.R., Agüera, A., 2007. Chemosphere 66 (6),
993–1002.
Gros, M., Petrović, M., Barceló, D., 2006. Anal Bioanal Chem 386 (4), 941–952.
Reist, M., Carrupt, P.-A., Francotte, E., Testa, B., 1998. Chem. Res. Toxicol 11 (12), 1521–1528.
Sweetman, S.C., 2009. Martindale: The complete drug reference, 36th ed. Pharmaceutical Press, London, P. 2397.
[email protected]
Quality control
Measured toxicity calculated
[email protected]
Waleed M.M. Mahmoud Ahmed thanks the Ministry of Higher
Education and Scientific Research of the Arab Republic of
Egypt (MHESR) and the German Academic Exchange Service
(DAAD) for their sponsorship and financial support (GERLS
program).
[email protected]