Studying the fate of the drug Chlorprothixene and its photo transformation products in the aquatic environment: Identification, assessment and priority setting by application of a combination of experiments and various in silico assessments.

Chlorprothixene (CPTX) is an antipsychotic drug of the thioxanthene class. Although it is widely used as a tranquillizer in psychiatry, anesthesiology, pediatrics, and in general medical practice, there is a gap in knowledge regarding its occurrence and fate in the environment. Therefore, we provide for the first-time data on the environmental fate and ecotoxicity of CPTX and its potential photo-transformations products (PTPs). Firstly, two standardized biodegradation tests (Closed Bottle test (CBT) and Manometric Respiratory test (MRT)) were performed to assess CPTX's environmental biodegradability. Then, its photodegradability was studied using Xenon and UV lamps. Effects of different conditions (initial drug concentration, pH, and temperature) were applied during UV-photodegradation. Subsequently, the time courses of CPTX and dissolved organic carbon (DOC) concentrations were monitored throughout the photodegradation tests. After that, high-resolution mass spectrometry was employed to elucidate the structures of the formed photo-transformation products (PTPs). In addition, biodegradation tests were performed for the photolytic mixtures to assess the biodegradability of the PTPs. Finally, the (eco)toxicity assessment for CPTX and its photolytic mixtures was predicted using different (quantitative) structure-activity relationship ((Q)SAR) software. CPTX was found to be not readily biodegradable in CBT and MRT. CPTX was not eliminated by irradiation with the Xenon lamp, however primarily eliminated using the UV-lamp. The CPTX elimination during UV-irradiation was faster at lower concentrations. CPTX UV-photodegradation was affected by pH value, while not affected by the temperature of the irradiated solution. 13 PTPs were detected in UV-photolysis mixtures. One additional product was detected in CPTX standard solution, and it was degraded simultaneously with CPTX during UV-irradiation. On one hand, Biodegradation assays revealed that UV-photolytic mixtures of CPTX, containing its PTPs, were not better biodegradable than CPTX itself. On the other hand, LC-MS analysis showed some PTPs which were eliminated after the biodegradation tests indicating possible biodegradability of these PTPs. This because those PTPs are present in low concentrations in the photolysis mixture and their effect can be hindered by the effect of CPTX and other non-biodegradable PTPs. QSAR analysis revealed that CPTX and some of its PTPs may have some human and/or eco-toxic properties. In conclusion, the release of CPTX into aquatic environments could be harmful. Therefore, further research focusing on CPTX and its PTPs are strongly recommended.

Biodegradation tests of mercaptocarboxylic acids, their esters, related divalent sulfur compounds and mercaptans.

Mercaptocarboxylic acids and their esters, a class of difunctional compounds bearing both a mercapto and a carboxylic acid or ester functional group, are industrial chemicals of potential environmental concern. Biodegradation of such compounds was systematically investigated here, both by literature search and by experiments (Closed Bottle Test OECD 301D and Manometric Respirometry Test OECD 301F). These compounds were found either readily biodegradable or at least biodegradable to a significant extent. Some related compounds of divalent sulfur were tested for comparison (mercaptans, sulfides, disulfides). For the two relevant monofunctional compound classes, carboxylic acids/esters and mercaptans, literature data were compiled, and by comparison with structurally similar compounds without these functional groups, the influence of COOH/COOR' and SH groups on biodegradability was evaluated. Thereby, an existing rule of thumb for biodegradation of carboxylic acids/esters was supported by experimental data, and a rule of thumb could be formulated for mercaptans. Concurrent to biodegradation, abiotic processes were observed in the experiments, rapid oxidative formation of disulfides (dimerisation of monomercaptans and cyclisation of dimercaptans) and hydrolysis of esters. Some problems that compromise the reproducibility of biodegradation test results were discussed.

Initial fate assessment of teratogenic drug trimipramine and its photo-transformation products - Role of pH, concentration and temperature.

Trimipramine (TMP) is an antidepressant drug used for the treatment of a variety of depressive states and other psychiatric disorders. It has been already detected in the aquatic environment. Currently, no further knowledge is available on fate and effects of TMP in the aquatic environment. Therefore, we studied the biodegradability of TMP and of its photolysis transformation products (PTPs) generated by irradiation with polychromatic UV light in aqueous solution. Different conditions including initial drug concentration, pH, and temperature were applied during TMP photolysis. Subsequently, the time courses of TMP and dissolved organic carbon (DOC) concentrations were monitored throughout the whole photo-degradation process. Then, high-resolution mass spectrometry was used to identify and elucidate the structures of the resulting PTPs. After that, the two standardized biodegradation tests, Closed Bottle test (CBT; OECD 301 D) and Manometric Respirometry test (MRT; OECD 301 F), were performed for TMP and its photolytic mixtures to assess the biodegradability of TMP and its PTPs. Finally, the toxicity of TMP and its photolytic mixtures was predicted using different quantitative structure activity relationship (QSAR) software. It was found that after 128 min of UV-irradiation, 91.8% of TMP at the initial concentration of 100 mg L-1 was eliminated with only 23.9% removal in the DOC. So, it can be pointed out that more than 65% of the degraded TMP is transformed to new non-mineralized PTPs. 14 new PTPs were detected in TMP's photolytic mixtures. Their supposed structures indicate that the proposed photo-transformation pathway is mainly by hydroxylation. The statistical analysis confirms that the differences in the degradation rates of TMP as a function of concentration, pH, and temperature are statistically significant in most cases investigated here. In biodegradation testing, TMP and its PTPs are classified as not readily biodegradable, while LC-MS analysis revealed some PTPs to be eliminated more than TMP itself. Results from QSAR analysis confirmed that some of the PTPs could be biodegradable, and revealed that some of the non-biodegradable PTPs may be human and/or eco-toxic, posing a risk to the environment. Our findings show that TMP under UV-irradiation could lead to the formation of some more easily biodegradable PTPs and some others toxic and non-biodegradable PTPs. Therefore, further studies should be conducted regarding the fate and effects of TMP and its PTPs elucidated in this study on human health and on the environment.

UV-photodegradation of desipramine: Impact of concentration, pH and temperature on formation of products including their biodegradability and toxicity.

Desipramine (DMI) is a widely used tricyclic antidepressant, and it is the major metabolite of imipramine (IMI) and lofepramine (LMI); IMI and LMI are two of the most commonly used tricyclic antidepressants. If DMI enters the aquatic environment, it can be transformed by the environmental bacteria or UV radiation. Therefore, photolysis of DMI in water was performed using a simulated sunlight Xenon-lamp and a UV-lamp. Subsequently, the biodegradability of DMI and its photo-transformation products (PTPs) formed during its UV photolysis was studied. The influence of variable conditions, such as initial DMI concentration, solution pH, and temperature, on DMI UV photolysis behavior was also studied. The degree of mineralization of DMI and its PTPs was monitored. A Shimadzu HPLC-UV apparatus was used to follow the kinetic profile of DMI during UV-irradiation; after that, ion-trap and high-resolution mass spectrometry coupled with chromatography were used to monitor and identify the possible PTPs. The environmentally relevant properties and selected toxicity properties of DMI and the non-biodegradable PTPs were predicted using different QSAR models. DMI underwent UV photolysis with first-order kinetics. Quantum yields were very low. DOC values indicated that DMI formed new PTPs and was not completely mineralized. Analysis by means of high-resolution mass spectrometry revealed that the photolysis of DMI followed three main photolysis pathways: isomerization, hydroxylation, and ring opening. The photolysis rate was inversely proportional to initial DMI concentration. The pH showed a significant impact on the photolysis rate of DMI, and on the PTPs in terms of both formation kinetics and mechanisms. Although temperature was expected to increase the photolysis rate, it showed a non-significant impact in this study. Results from biodegradation tests and QSAR analysis revealed that DMI and its PTPs are not readily biodegradable and that some PTPs may be human and/or eco-toxic, so they may pose a risk to the environment.

Identification of phototransformation products of thalidomide and mixture toxicity assessment: an experimental and quantitative structural activity relationships (QSAR) approach.

The fate of thalidomide (TD) was investigated after irradiation with a medium-pressure Hg-lamp. The primary elimination of TD was monitored and structures of phototransformation products (PTPs) were assessed by LC-UV-FL-MS/MS. Environmentally relevant properties of TD and its PTPs as well as hydrolysis products (HTPs) were predicted using in silico QSAR models. Mutagenicity of TD and its PTPs was investigated in the Ames microplate format (MPF) aqua assay (Xenometrix, AG). Furthermore, a modified luminescent bacteria test (kinetic luminescent bacteria test (kinetic LBT)), using the luminescent bacteria species Vibrio fischeri, was applied for the initial screening of environmental toxicity. Additionally, toxicity of phthalimide, one of the identified PTPs, was investigated separately in the kinetic LBT. The UV irradiation eliminated TD itself without complete mineralization and led to the formation of several PTPs. TD and its PTPs did not exhibit mutagenic response in the Salmonella typhimurium strains TA 98, and TA 100 with and without metabolic activation. In contrast, QSAR analysis of PTPs and HTPs provided evidence for mutagenicity, genotoxicity and carcinogenicity using additional endpoints in silico software. QSAR analysis of different ecotoxicological endpoints, such as acute toxicity towards V. fischeri, provided positive alerts for several identified PTPs and HTPs. This was partially confirmed by the results of the kinetic LBT, in which a steady increase of acute and chronic toxicity during the UV-treatment procedure was observed for the photolytic mixtures at the highest tested concentration. Moreover, the number of PTPs within the reaction mixture that might be responsible for the toxification of TD during UV-treatment was successfully narrowed down by correlating the formation kinetics of PTPs with QSAR predictions and experimental toxicity data. Beyond that, further analysis of the commercially available PTP phthalimide indicated that transformation of TD into phthalimide was not the cause for the toxification of TD during UV-treatment. These results provide a path for toxicological assessment of complex chemical mixtures and in detail show the toxic potential of TD and its PTPs as well as its HTPs. This deserves further attention as UV irradiation might not always be a green technology, because it might pose a toxicological risk for the environment in general and specifically for water compartments.

Aquatic photochemistry, abiotic and aerobic biodegradability of thalidomide: identification of stable transformation products by LC-UV-MS(n).

Thalidomide (TD), besides being notorious for its teratogenicity, was shown to have immunomodulating and anti-inflammatory activities. This is why recently TD became a promising drug for the treatment of different cancers and inflammatory diseases. Yet nothing is known about the environmental fate of TD, which therefore was assessed experimentally and by in silico prediction programs (quantitative structure activity relationship (QSAR) models) within this study. Photolytic degradation was tested with two different light sources (medium-pressure mercury lamp; xenon lamp) and aerobic biodegradability was investigated with two OECD tests (Closed Bottle test (CBT), Manometric Respirometry test (MRT)). An additional CBT was performed for TD samples after 16 min of UV-photolysis. The primary elimination of TD was monitored and the structures of its photo-, abiotic and biodegradation products were elucidated by HPLC-UV-Fluorescence-MS(n). Furthermore, elimination of dissolved organic carbon was monitored in the photolysis experiment. LC-MS revealed that new photolytic transformation products (TPs) were identified, among them two isomers of TD with the same molecular mass. These TPs were different to the products formed by biodegradation. The experimental findings were compared with the results obtained from the in silico prediction programs where e.g. a good correlation for TD biodegradation in the CBT was confirmed. Moreover, some of the identified TPs were also structurally predicted by the MetaPC software. These results demonstrate that TD and its TPs are not readily biodegradable and not fully mineralized by photochemical treatment. They may therefore pose a risk to the aquatic environment due to the pharmacological activity of TD and unknown properties of its TPs. The applied techniques within this study emphasize the importance of QSAR models as a tool for estimating environmental risk assessments.

Oxidation-coagulation of ß-blockers by K2FeVIO4 in hospital wastewater: assessment of degradation products and biodegradability.

This study investigated the degradation of atenolol, metoprolol and propranolol beta-blockers by ferrate (K2FeO4) in hospital wastewater and in aqueous solution. In the case of hospital wastewater, the effect of the independent variables pH and [Fe(VI)] was evaluated by means of response surface methodology. The results showed that Fe(VI) plays an important role in the oxidation-coagulation process, and the treatment of the hospital wastewater led to degradations above 90% for all the three ß-blockers, and to reductions of aromaticity that were close to 60%. In addition, only 17% of the organic load was removed. In aqueous solution, the degradation of the ß-blockers atenolol, metoprolol and propranolol was 71.7%, 24.7% and 96.5%, respectively, when a ratio of 1:10 [ß-blocker]:[Fe(VI)] was used. No mineralization was achieved, which suggests that there was a conversion of the ß-blockers to degradation products identified by liquid chromatography/mass spectrometry tandem. Degradation pathways were proposed, which took account of the role of Fe(VI). Furthermore, the ready biodegradability of the post-process samples was evaluated by using the closed bottle test, and showed an increase in biodegradability. The use of the ferrate advanced oxidation technology seems to be a useful means of ensuring the remediation of hospital and similar wastewater.

Photolysis of sulfamethoxypyridazine in various aqueous media: aerobic biodegradation and identification of photoproducts by LC-UV-MS/MS.

Sulfonamides are one of the most frequently used antibiotics worldwide. Therefore, mitigation processes such as abiotic or biotic degradation are of interest. Photodegradation and biodegradation are the potentially significant removal mechanisms for pharmaceuticals in aquatic environments. The photolysis of sulfamethoxypyridazine (SMP) using a medium pressure Hg-lamp was evaluated in three different media: Millipore water pH 6.1 (MW), effluent from sewage treatment plant pH 7.6 (STP), and buffered demineralized water pH 7.4 (BDW). Identification of transformation products (TPs) was performed by LC-UV-MS/MS. The biodegradation of SMP using two tests from the OECD series was studied: Closed Bottle test (OECD 301 D), and Manometric Respirometry test (OECD 301 F). In biodegradation tests, it was found that SMP was not readily biodegradable so it may pose a risk to the environment. The results showed that SMP was removed completely within 128 min of irradiation in the three media, and the degradation rate was different for each investigated type of water. However, dissolved organic carbon (DOC) was not removed in BDW and only little DOC removal was observed in MW and STP, thus indicating the formation of TPs. Analysis by LC-UV-MS/MS revealed new TPs formed. The hydroxylation of SMP represents the main photodegradation pathway.

Captopril and its dimer captopril disulfide: photodegradation, aerobic biodegradation and identification of transformation products by HPLC-UV and LC-ion trap-MS(n).

In some countries effluents from hospitals and households are directly emitted into open ditches without any further treatment and with very little dilution. Under such circumstances photo- and biodegradation in the environment can occur. However, these processes do not necessarily end up with the complete mineralization of a chemical. Therefore, the biodegradability of photoproduct(s) by environmental bacteria is of interest. Cardiovascular diseases are the number one cause of death globally. Captopril (CP) is used in this study as it is widely used in Egypt and stated as one of the essential drugs in Egypt for hypertension. Three tests from the OECD series were used for biodegradation testing: Closed Bottle test (CBT; OECD 301 D), Manometric Respirometry test (MRT; OECD 301 F) and the modified Zahn-Wellens test (ZWT; OECD 302 B). Photodegradation (150 W medium-pressure Hg-lamp) of CP was studied. Also CBT was performed for captopril disulfide (CPDS) and samples received after 64 min and 512 min of photolysis. The primary elimination of CP and CPDS was monitored by LC-UV at 210 nm and structures of photoproducts were assessed by LC-UV-MS/MS (ion trap). Analysis of photodegradation samples by LC-MS/MS revealed CP sulfonic acid as the major photodegradation product of CP. No biodegradation was observed for CP, CPDS and of the mixture resulting from photo-treatment after 64 min in CBT. Partial biodegradation in the CBT and MRT was observed in samples taken after 512 min photolysis and for CP itself in MRT. Complete biodegradation and mineralization of CP occurred in the ZWT.

Development and validation of a stability-indicating RP-HPLC method for the determination of paracetamol with dantrolene or/and cetirizine and pseudoephedrine in two pharmaceutical dosage forms.

A stability-indicating reversed-phase high-performance liquid chromatography (RP-HPLC) method has been developed which can separate and accurately quantitate paracetamol, dantrolene, cetirizine and pseudoephedrine. The method was successfully validated for the purpose of conducting stability studies of the four analytes in quality control (QC) laboratories. The stability-indicating capability of the method was demonstrated by adequate separation of these four analytes from all the degradant peaks. A gradient mobile phase system consisting of (A) 50 mmol L(-1) sodium dihydrogen phosphate, 5 mmol L(-1) heptane sulfonic acid sodium salt, pH 4.2 and (B) acetonitrile was used with Discovery reversed-phase HS C(18) analytical column (250 mm x 4.6 mm i.d., 5 microm particle size). Quantitation was achieved with UV detection at 214 nm, based on peak area. The proposed method was validated and successfully applied for the analysis of pharmaceutical formulations and laboratory-prepared mixtures containing the two multicomponent combinations.

New validated liquid chromatographic and chemometrics-assisted UV spectroscopic methods for the determination of two multicomponent cough mixtures in syrup.

Multivariate spectrophotometric calibration and liquid chromatographic (LC) methods were applied to the determination of 2 multicomponent mixtures containing diprophylline, guaiphenesin, methylparaben, and propylparaben (Mixture 1), or clobutinol, orciprenaline, saccharin sodium, and sodium benzoate (Mixture 2). For the multivariate spectrophotometric calibration methods, principal component regression (PCR) and partial least-squares regression (PLS-1), a calibration set of the mixtures consisting of the components of each mixture was prepared in 0.1 M HCl. Analytical figures of merit such as sensitivity, selectivity, limit of quantitation, and limit of detection were determined for both PLS-1 and PCR. The LC separation was achieved on a reversed-phase C18 analytical column by using isocratic elution with 20 mM potassium dihydrogen phosphate, pH 3.3-acetonitrile (55 + 45, v/v) as the mobile phase and UV detection at 260 and 220 nm for Mixture 1 and Mixture 2, respectively. The proposed methods were validated and successfully applied to the analysis of pharmaceutical formulations and laboratory-prepared mixtures containing the 2 multicomponent combinations.

HPLC and chemometrics-assisted UV-spectroscopy methods for the simultaneous determination of ambroxol and doxycycline in capsule.

High-performance liquid chromatography (HPLC) and multivariate spectrophotometric methods are described for the simultaneous determination of ambroxol hydrochloride (AM) and doxycycline (DX) in combined pharmaceutical capsules. The chromatographic separation was achieved on reversed-phase C(18) analytical column with a mobile phase consisting of a mixture of 20mM potassium dihydrogen phosphate, pH 6-acetonitrile in ratio of (1:1, v/v) and UV detection at 245 nm. Also, the resolution has been accomplished by using numerical spectrophotometric methods as classical least squares (CLS), principal component regression (PCR) and partial least squares (PLS-1) applied to the UV spectra of the mixture and graphical spectrophotometric method as first derivative of the ratio spectra ((1)DD) method. Analytical figures of merit (FOM), such as sensitivity, selectivity, analytical sensitivity, limit of quantitation and limit of detection were determined for CLS, PLS-1 and PCR methods. The proposed methods were validated and successfully applied for the analysis of pharmaceutical formulation and laboratory-prepared mixtures containing the two component combination.

Development and validation of chemometrics-assisted spectrophotometry and liquid chromatography methods for the simultaneous determination of the active ingredients in two multicomponent mixtures containing chlorpheniramine maleate and phenylpropanolamine hydrochloride.

Multivariate spectrophotometric calibration and liquid chromatography (LC) methods were used for the simultaneous determination of the active ingredients in 2 multicomponent mixtures containing chlorpheniramine maleate and phenylpropanolamine hydrochloride with ibuprofen and caffeine (mixture 1) or with propyphenazone (mixture 2). For the multivariate spectrophotometric calibration methods, principal component regression (PCR) and partial least squares (PLS-1), a calibration set of the mixtures consisting of the components of each mixture was prepared in distilled water. A leave-1-out cross-validation procedure was used to find the optimum numbers of latent variables. Analytical parameters such as sensitivity, selectivity, analytical sensitivity, limit of quantitation, and limit of detection were determined for both PLS-1 and PCR. The LC method depends on the use of a cyanopropyl column with the mobile phase acetonitrile-12 mM ammonium acetate, pH 5.0 (25 + 75, v/v), for mixture 1 or acetonitrile-10 mM potassium dihydrogen phosphate, pH 4.7 (45 + 55, v/v), for mixture 2; the UV detector was set at 212 nm. In spite of the presence of a high degree of spectral overlap of these components, they were rapidly and simultaneously determined with high accuracy and precision, with no interference from the matrix excipients. The proposed methods were successfully applied to the analysis of pharmaceutical formulations and laboratory-prepared mixtures containing the 2 multicomponent combinations.

High performance liquid chromatographic determination of etofibrate and its hydrolysis products.

High performance liquid chromatographic (HPLC) method is presented for the determination of etofibrate (EF) and its hydrolysis products. The method was based on HPLC separation of EF from its hydrolysis products using cyanopropyl column at ambient temperature with mobile phase consisting of acetonitrile-10 mM potassium dihydrogen phosphate, pH was adjusted to 4.1 using phosphoric acid (50:50, v/v). Quantitation was achieved with UV detection at 221 nm based on peak area. The flow rate was 1.5 ml min(-1). The proposed method was used to investigate the kinetics of acidic hydrolysis process of EF at different temperatures and the apparent pseudo first-order rate constant, half-life and activation energy were calculated. The kinetics of alkaline hydrolysis process of EF using 0.01 M sodium hydroxide at different temperatures cannot be studied as the drug is rapidly hydrolyzed in alkaline medium. The pH-rate profile of hydrolysis of EF in Britton-Robinson buffer solutions within the pH range 2-10 were studied.