Environ Chem Lett (2005) 2:195198

DOI 10.1007/s10311-004-0096-1

ORIGINAL PAPER

M. Brigante M. DellaGreca L. Previtera

M. Rubino F. Temussi

Degradation of hydrochlorothiazide in water

Received: 18 March 2004 / Accepted: 22 November 2004 / Published online: 21 January 2005

Springer-Verlag 2005

Abstract The photolytic decomposition of hydrochlorothiazide in water was investigated. Irradiation of the drug
was performed in pure water and sewage treatment plant
water with a solar simulator and by direct sunlight. Here
we show that three photoproducts were obtained. Two of
them were formed in yields much higher than 10%. An
environmental risk assessment of hydrochlorothiazide
should take into account also these photoproducts in a
complete analytical and ecotoxicological evaluation.
Keywords Hydrochlorothiazide Drug Photodegradation Phototransformation Water

Introduction
The occurrence of pharmaceutical chemicals in natural
waters has been extensively reported and a growing interest is addressed to their ecological impact (Daughton
and Ternes 1999; Halling-Sorensen et al. 1998; Kmmerer
2001). In 2001, the Scientific Committee on Toxicity,
Ecotoxicity and the Environment (CSTEE 2001) has
recommended that attention should also be paid to degradation and reaction products of pharmaceuticals.
Hydrochlorothiazide (6-chloro-3,4-dihydro-2H-1,2,4benzothiadiazine-7-sulfonamide1,1-dioxide) (1a) is a diuretic and antihypertensive and it is supplied as tablets for
oral use. It is a white, or practically white, crystalline
powder, which is slightly soluble in water, but freely
soluble in sodium hydroxide solution. It is well known
M. Brigante
Dipartimento di Scienze della Vita,
II Universit di Napoli,
Via Vivaldi 43, I-81100 Caserta, Italy
M. DellaGreca L. Previtera M. Rubino F. Temussi ())
Dipartimento di Chimica Organica e Biochimica,
Universit Federico II,
Via Cinthia 4, I-80126 Naples, Italy
e-mail: ftemussi@unina.it
Tel.: +39081674471
Fax: +39081674393

that hydrochlorothiazide is not metabolised and at least

61% of the oral dose is eliminated by the kidney unchanged within 24 h (OGrady et al. 1999).
In a recent study, a survey was done in the Po and
Lambro Rivers in Italy to check the presence of therapeutic drugs in the aquatic environment and hydrochlorothiazide has been detected in concentrations ranging
from 10 to 250 ng/l (Calamari et al. 2003). As stated by
the CSTEE, an environmental risk assessment for a
pharmaceutical drug should take into account also the
transformation products, especially those obtained in
percentages greater than 10% (CSTEE 2001). In this light,
we decided to investigate the photostability of hydrochlorothiazide in water to obtain information on the
phototransformation products that could be formed in
surface waters and sewage treatment plant (STP) waters.
Previous studies on the photostability of hydrochlorothiazide are present in literature. Tamat and Moore (1983)
studied its photocatalytic decomposition. In the work the
drug is reported to decompose upon irradiation with near
UV-light (>310 nm) in methanol and aqueous solutions.
The source of UV light employed was a medium-pressure
mercury vapor lamp with glass filter. The primary photoprocesses were photodehalogenation and photohydrolysis. In the photolysis in the aqueous (5% methanol) solution the dominant photoproduct was the hydrolysed and
dechlorinated 2b, and only small amounts of 2a (4-amino6-chloro-1,3-benzenedisulfonamide) were found (Fig. 1).
The products were characterised by gas chromatography
mass spectrometry (GCMS) using synthesised standards.
Furthermore, the mechanism of photolysis involving a
radical cation formation was discussed. Revelle et al.
(1997) have re-investigated the photolytic decomposition
in methanol. In this case, the source of the ultraviolet
(UV)-light employed was a UV-A fluorescent lamp with
wavelengths ranging from 300 to 400 nm. Photodehalogenation was reported to be the primary degradation
process in which the chlorine of hydrochlorothiazide is
replaced by hydrogen (1b) or by a methoxyl group (1c)
from the methanol solvent (Fig. 1). They did not detect
the hydrolysed product 2a, while they observed the for-

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periments. The heteronuclear chemical shift correlations

were determined by heteronuclear multiple quantum coherence (HMQC) and heteronuclear multiple bond coherence (HMBC) pulse sequences.
Irradiation of hydrochlorothiazide in water

Fig. 1 Hydrochlorothiazide and its photoproducts

mation of 2c and a hydrochlorothiazide photodehydrogenation process that leaded to chlorothiazide (3a) and
derivatives 3b, 3c (Fig. 1). The different photoproduct
distribution for hydrochlorothiazide was attributed to the
UV-A lamps employed and the method of product detection and identification. 1b and 2a were the main
products obtained by Ulvi and Tammilehto (1989) when
hydrochlorothiazide was irradiated in ethanolic solution
with a high-pressure mercury lamp. This report describes
the photochemical transformation of hydrochlorothiazide
in water when irradiated by sunlight.

Experimental
Equipment and methods
High-performance liquid chromatography (HPLC) experiments were carried out on an Agilent 1100 HPLC
system equipped with an ultraviolet (UV) detector. Irradiations were performed using a 150-W solar simulator
equipped with a Xenon lamp with a spectral output 200
2,400 nm using a filter to simulate irradiation at the earth
surface (Oriel Instruments). Elemental analysis was performed on a 2400 Series II CHNS/O Analyzer (PerkinElmer). Mass spectra were recorded using a Finnigan
LCQ equipped with an electrospray ionisation (ESI)
probe operating in negative ion mode. The scan range was
802,000 m/z.
Nuclear magnetic resonance (NMR) spectra were
recorded at 500 MHz for [1H] and 125 MHz for [13C] on a
Fourier Transform NMR Varian 500 Unity Inova spectrometer. The carbons multiplicity was evidenced by
distortionless enhancement by polarisation transfer
(DEPT) experiments. The proton couplings were evidenced by 1H1H correlation spectroscopy (COSY) ex-

In a typical procedure a suspension of hydrochlorothiazide (1a) (100 mM) in distilled water was irradiated at
room temperature for 200 h by a 150-W solar simulator.
To follow the irradiation experiment, an aliquot was
withdrawn at various times, concentrated and the residue
dissolved in methanol, and injected in a HPLCUV system. The column used was a RP-18 column (Phenomenex
HYDRO RP-18, 4 m, 250 mm4.5 mm. The column was
eluted with a mixture of A (H2O containing 1% acetic
acid) B (MeOH containing 1% acetic acid) 90:10 (v/v),
detection was at 260 nm and the flow rate was 0.7 ml/min.
Experiments in the same irradiation conditions were
run also in distilled water under argon atmosphere, and in
water of a sewage treatment plant (STP). STP water was
obtained from Mercato S. Severino treatment plant
(Salerno, Italy).
Irradiation of hydrochlorothiazide in distilled water
(preparative scale)
A solution of hydrochlorothiazide (1a) (0.7 mM) in distilled water was irradiated at room temperature for 200 h
by a 150-W solar simulator. The water was evaporated to
dryness. The mixture (100 mg) separated by silica gel
flash column chromatography eluting with hexaneethylic etheracetone 3:3:4, yielded two fractions: A (70 mg)
and B (26 mg). Fraction A contained hydrochlorothiazide
and compound 2a. Compound 2a (29 mg) was separated
from hydrochlorothiazide on a preparative TLC plate
(0.5 mm) eluting with hexaneethylic etheracetone 3:3:4
(TLC: thin layer chromatography). Fraction B was purified from reverse phase (RP)HPLC (for experimental
conditions see above) and compounds 1d (20 mg) and 2d
(2 mg) were obtained.
Spectral data of photoproducts
Compound 2a: ESI-MS: m/z 278 [M-1]. 1H NMR
(CD3OD) d 8.34 (1H, s, H-2), 6.97 (1H, s, H-5). 13C NMR
(CD3OD) d 150.5 (C-4), 137.0 (C-1), 132.1 (C-2), 128.2
(C-6), 123.0 (C-3), 119.2 (C-5).
Compound 1d: Elemental analysis (found: C, 30.28; H,
3.44; N, 15.22; O, 28.70; S, 23.12; calcd: C, 30.24; H,
3.50; N, 15.18; O, 28.78; S, 23.06). ESI-MS: m/z 278 [M1]. 1H NMR (CD3OD) d 7.92 (1H, s, H-8), 6.24 (1H, s, H5), 4.70 (2H, s, H-3). 13C NMR (CD3OD) d 160.0 (C-6),
150.0 (C-40 ), 127.0 (C-8), 120.2 (C-7), 114.6 (C-80 ), 101.9
(C-5), 56.0 (C-3).

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Fig. 2 Photodegradation of hydrochlorothiazide (1a) in water.

Analysis by HPLC showing the increase of the relative concentration of photoproducts (1d, 2a, 2d) with time of exposure

Compound 2d: Elemental analysis (found: C, 27.15; H,

3.73; N 15.80; O, 30.15; S, 24.06; calcd: C, 27.09; H,
3.65, N, 15.87; O, 30.08; S, 24.10). ESI-MS: m/z 266 [M1]. 1H NMR (CD3OD) d 8.07 (1H, s, H-2), 6.18 (1H, s, H5). 13C NMR (CD3OD) d 152.6 (C-6), 145.6 (C-4), 131.9
(C-2), 119.5 (C-1), 116.5 (C-5),116.2 (C-3).

Results and discussion

In order to understand the fate of hydrochlorothiazide
when exposed to sunlight in surface and sewage treatment
plant waters, we investigated the photochemical behaviour in water (Fig. 2). For the purpose of simulating the
photodegradation under environmental conditions, hydrochlorothiazide has been suspended in pure water and
irradiated at different times with a solar simulator. The
experiment has been followed injecting an aliquot of the
water sample withdrawn at various times in a HPLCUV
system. After 5 h, we observed a new peak in the HPLC
chromatogram (Fig. 2), this peak grew with the irradiation
time and became constant after 30 h of irradiation. Only
after 30 h we observed the appearance of two other peaks
in minor amounts and after 200 h it was evident that three
main photoproducts (2a, 1d, 2d) were present.
To isolate and characterise the three photoproducts
obtained, we performed an irradiation experiment in a
preparative scale. Hydrochlorothiazide (0.7 mM), suspended in pure water, was irradiated for 200 h with the
solar simulator. The irradiated solution was evaporated to
dryness and the mixture was submitted to flash chromatography yielding three compounds along with hydrochlorothiazide. The three photoproducts were further purified by TLC and HPLC. The most abundant compound
has been determined to be 4-amino-6-chloro-1,3-benzenedisulfonamide (2a). NMR data were consistent with
literatura data (Revelle et al 1997). Further structural information was obtained by ESI-MS analysis. Thus, the

photolysis of hydrochlorothiazide in pure water leads to

hydrolysis of the thiadazine ring as dominant product.
Compound 2a is known as a degradation product of hydrochlorothiazide (Franolic et al 2001), so we performed
an experiment in the dark to check the influence of light
in this process. After 200 h in the dark, hydrochlorothiazide was quantitatively recovered, indicating that light is
necessary in the photohydrolysis of hydrochlorothiazide.
Compound 1d, formed in 15% after 200 h irradiation,
was identified as 6-hydroxy-3,4-dihydro-2H-1,2,4-benzothiadazine-7-sulfonamido-1,1-dioxide. To the best of
our knowledge, this compound was only suggested as
intermediate in the mechanism of photodegradaton by
Tamat and Moore (1983), but it was never isolated and
described before now. In the ESI-MS spectrum the molecular peak at 278 was present in agreement with the
molecular formula C7H9N3O5S2. The 1H NMR spectrum
shows two aromatic protons, as singlets at d 6.24 and
7.92, and two protons of a methylene as a singlet at d
4.70.
In the 13C NMR spectrum seven carbon signals can be
identified. Three protonated carbons at d 127.0, 101.9 and
56.0 and four aromatic quaternary carbons at d 160.0,
150.0, 120.2, and 114.6 were detectable. All these data
are consistent with the structure proposed. This compound
was formed by photosubstitution from hydrochlorothiazide in a process where the chlorine is replaced by OH
from the solvent. To verify that oxygen was not involved
in this process, we irradiated the hydrochlorothiazide
suspension under argon atmosphere. After 200 h, we
observed the formation of compound 1d in 50% yield.
The structure 4-amino-6-hydroxy-1,3-benzenedisulfonamide was attributed to compound 2d (5%). The molecular peak at 266, along with the elemental analysis defined the molecular formula C6H9N3O5S2. The 1H NMR
spectrum exhibited two aromatic protons, as singlets at d
6.18 and 8.07. In the 13C NMR spectrum six carbon
signals were present: two protonated carbons at d 131.9,
116.5 and four aromatic quaternary carbons at d 152.6,
145.6, 119.5 and 116.2.
All compounds were used as standards to evaluate the
phototransformation yields by HPLC analysis. The yields
were also confirmed by 1H NMR integration analysis of
the mixture after irradiation. The yields of photoproduct
2a, 1d and 2d after 200 h, were 35, 15 and 5%, respectively.
To verify the phototransformation in a simulated
aquatic environment an irradiation experiment was also
performed suspending hydrochlorothiazide in sewage
treatment plant (STP) water. The same photoproducts 2a,
1d, 2d were obtained after 200 h. The only difference
observed in this case was the little yield of compound 2d
(1%). Finally, we performed an experiment irradiating the
hydrochlorothiazide suspension (pure water) under sunlight for 5 days in January, surprisingly hydrochlorothiazide was almost completely transformed and the main
product was compound 1d (75% yield).
The occurrence of the same three photoproducts in all
irradiation conditions tested suggests a unique photolysis

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pathway for hydrochlorothiazide in water, where the main

processes are the photohydrolysis of the thiadazine ring
and the photosubstitution of the chlorine with the hydroxyl group.

Conclusion
Hydrochlorothiazide was irradiated in biomimetic conditions for 200 h leading to three photoproducts that were
isolated and characterised. Two of them were isolated in
percentage significantly higher than 10%. For an environmental risk assessment of hydrochlorothiazide, further
studies on the distribution of hydrochlorothiazide including its phototransformation products in the aquatic
environment as well as eco-toxicological data are needed.
Acknowledgements This work was supported by MIUR (Ministero
Istruzione Universit e Ricerca) in the frame of PRIN 2002 (Progetto Ricerca Interesse Nazionale). NMR experiments were performed at CIMCF (Centro Interdipartimentale di metodologie
chimico-fisiche) on a Varian 500 Inova spectrometer of Consortium
INCA (Interuniversitario Nazionale Consorzio Ambiente) (Cluster
11-A, Project P0, L.488). The authors wish also to thank Dr. G.
Giacometti.

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