PENETRATION OF SOLAR UV IRRADIATION IN COASTAL LAGOONS OF THE SOUTHERN BALTIC SEA AND ITS EFFECT ON PHYTOPLANKTON COMMUNITIES

Abstract Direct measurements of solar UV irradiance at the surface and in the water column were made in several coastal lagoons near the islands of Hiddensee and Rügen in spring and summer 1993 using an internally frosted quartz sphere at the end of a 10 m quartz fiber cable connected to a double monochromator spectroradiometer. The measurements showed a nearly uniform vertical distribution of the absorption coefficient in the whole water column due to effective mixing, shallow depths and little temperature and salinity density stratifications. There was an increase in the absorption coefficient at 310 nm from 5 m⁻¹ in the outer lagoons to 12 m⁻¹ in the open inner lagoons and up to 29 m⁻¹ in the inner lagoons without water exchange with the Baltic Sea as a result of different amounts of UV-absorbing substances such as yellow substance (Gelbstoff) and seston. Depending on the specific concentration of the absorbents, the solar UV-B irradiance decreases to 1% after penetration into the water column between 0.2 m (inner lagoons without water exchange), 0.4 m (inner lagoons with water exchange), and 1 m (outer lagoons), whereas UV-A penetrates on the order of several meters. As phytoplankton populates the top layers of the water column it may be affected by increased solar UV irradiation even in these turbid coastal waters.     

Spatial and temporal variations of the inherent and apparent optical properties in a shallow coastal lake

Apparent and inherent optical properties in the coastal lagoon of Fogliano were measured in three seasonal surveys in 2002. Irradiance data from in situ measurements of ultraviolet and visible wavebands permitted to estimate the related attenuation coefficients. Laboratory extinction measurements on filtered (0.22 microm) and unfiltered water samples were also performed. The integrated approach between in situ and laboratory measurements allowed the determination of the role of the suspended and dissolved matter in the attenuation and extinction of ultraviolet and visible radiation within the water column. As noted, the impact of the suspended and dissolved matter on the lake optical quality was influenced by wind resuspension of particulate matter: the relative role of dissolved matter in the absorption of UV and visible radiation was prevailing at low wind velocity conditions (less than 2.2 m s(-1)), while at high wind velocities (3.9 m s(-1)), particulate matter resuspension strongly influenced the attenuation and the extinction measurements. The extinction in the analyzed wavelengths of filtered and unfiltered water samples and the in situ irradiance measurements allowed us to define new optical parameters and important correlations with limnological and classical optical measurements. By sampling at high spatial resolution (18 stations in 4 km2), it was possible to evidence a spatial gradient of the optical and limnological properties, these distributions showed a consistent pattern in all three surveys, and were important for the characterization of the chromophoric dissolved organic matter that was estimated with the spectral slope of the extinction curve spectra. A relatively higher spectral slope was found in the southern basin with respect to the northern, where the maximum values of the attenuation coefficients and limnological parameters were found. These results suggest different sources of dissolved organic matter and/or a different level of photobleaching.     

MODEL FOR THE GLOBAL IRRADIANCE OF THE SOLAR BIOLOGICALLY-EFFECTIVE ULTRAVIOLET-RADIATION ON INCLINED SURFACES

The body surface area of man is the relevant receiving surface for solar UV radiation. To consider this body surface geometry, the biologically-effective UV radiation of the solar global radiation was measured. This was done at 26 differently aligned measuring points whose orientation was determined by the angle of inclination (vertical) and the azimuth (horizontal). Approximately eight hundred sets of measurement series were carried out at 33 different sites. A simple model, developed from the data obtained, made it possible to calculate relative irradiance as a function of the angle of inclination and the ground reflection (UV albedo). Thus relative risk of solar UV exposure to different regions of the body can be assessed. In addition to this, if the irradiance on a horizontal plane (measured or calculated by a corresponding model) is taken into consideration, the absolute values for UV irradiance on tilted planes can be determined.     

THE INACTIVATION OF A NATURAL POPULATION OF COLIFORM BACTERIA BY SUNLIGHT

Abstract— The 'natural purification' of water, processes by which pathogenic organisms disappear from contaminated waters, is of great importance regarding water resources. The relative importance of the various processes which remove undesirable organisms is yet to be determined. Wastewater was exposed to sunlight by placing 120 m/samples in UV transparent or absorptive cylindrical vessels which were then placed in a 50/ container filled with water from a lagoon system, thus simulating actual exposure conditions in the lagoon system. Samples were exposed to natural sunlight for 3 days and received different wavelengths of sunlight, depending upon the presence of predetermined filters. The dynamics of the coliform population was observed. It was noted that coliform bacteria demonstrated a significant growth potential when held in darkness or during the night. An entire day of full unfiltered sunlight was a potent bacteriocidal agent, reducing coliform survival to less than 0.01% of the initial value. Removing the shorter wavelengths (Λ. < 325 nm) by optical filters or by the optical properties of the wastewater itself still left components of sunlight capable of killing coliforms. Our studies support the concept that solar UV radiation makes a substantial contribution to the natural purification of water.     

Damage to DNA in bacterioplankton: a model of damage by ultraviolet radiation and its repair as influenced by vertical mixing

A model of UV-induced DNA damage in oceanic bacterioplankton was developed and tested against previously published and novel measurements of cyclobutane pyrimidine dimers (CPD) in surface layers of the ocean. The model describes the effects of solar irradiance, wind-forced mixing of bacterioplankton and optical properties of the water on net DNA damage in the water column. The biological part includes the induction of CPD by UV radiation and repair of this damage through photoreactivation and excision. The modeled damage is compared with measured variability of CPD in the ocean: diel variation in natural bacterioplankton communities at the surface and in vertical profiles under different wind conditions (net damage as influenced by repair and mixing); in situ incubation of natural assemblages of bacterioplankton (damage and repair, no mixing); and in situ incubation of DNA solutions (no repair, no mixing). The model predictions are generally consistent with the measurements, showing similar patterns with depth, time and wind speed. A sensitivity analysis assesses the effect on net DNA damage of varying ozone thickness, colored dissolved organic matter concentration, chlorophyll concentration, wind speed and mixed layer depth. Ozone thickness and mixed layer depth are the most important factors affecting net DNA damage in the mixed layer. From the model, the total amplification factor (TAF; a relative measure of the increase of damage associated with a decrease in ozone thickness) for net DNA damage in the euphotic zone is 1.7, as compared with 2.1-2.2 for irradiance weighted for damage to DNA at the surface.     

UVB-INDUCED SUPPRESSION OF THE MIXED EPIDERMAL CELL LYMPHOCYTE REACTION IS CRITICALLY DEPENDENT ON IRRADIANCE

The mixed epidermal cell lymphocyte reaction (MECLR) is a commonly used method to study the effects of ultraviolet B (UVB) radiation on the skin immune system. In UVB experiments dosimetry is very important. The influence of irradiance on the MECLR was studied in vitro using Philips FS40 lamps with variable UV intensities. Irradiation of isolated epidermal cells with high irradiance impaired the alloactivating capacity more than irradiation with low irradiance. In vivo, the influence of long-term UVB exposure on the MECLR was studied by treating normal healthy volunteers with suberythemagenic doses of UVB thrice weekly during 4 weeks. The first set of experiments, using low irradiance Sylvania UV-21 F75/85 W lamps, resulted in a decrease of MECLR responses of 83.1%. In the second set of experiments performed a year later, employing an identical protocol except for the use of high irradiance Waldmann UV-21 F85/100 W lamps, an increase of MECLR responses of 99.7% was observed. Volunteers of both sets of experiments received equal doses of UVB. In conclusion, this study shows that in vitro UVB-induced suppression of the MECLR is critically dependent on irradiance and therefore might explain contradictory results described in the literature. The in vivo data suggest that, comparable to the in vitro experiments, irradiance may influence the effects of UVB irradiation in vivo. Further experiments should prove whether this is indeed the case.     

POTENTIAL ERRORS IN THE USE OF CELLULOSE DIACETATE AND MYLAR FILTERS IN UV-B RADIATION STUDIES

Abstract— The increase in UV-B radiation(290–320 nm) penetrating to the earth's surface as a result of the chemical depletion of the stratospheric ozone layer is an important environmental concern. In most studies using artificial UV-B sources, the determination of enhanced UV-B radiation effects on plants relies on equivalent UV-A radiation(320–400 nm) from the experimental UV-B fluorescent lamp source, filtered with either cellulose diacetate (CA) to create UV-B treatments, or with type S Mylar or polyester (PE) to create controls (no UV-B). The spectral irradiance in the UV-A was measured in the dark below lamps at two daily UV-B irradiance levels (14.1 and 10.7 W m^-2) with CA and PE at two ages. Highly significant differences in UV-A radiation (P 0.01) were measured below the treatment/control pairs at both fluence rates and filter ages. Filter aging was observed, which reduced the UV-A irradiance, especially for PE. The total daily ambient UV-A irradiance was also determined in the glasshouse at three seasons: the fall equinox, summer and winter, from which the total daily UV-A (lamp + ambient) irradiances were calculated. The addition of low to moderate ambient irradiance removed the treatment/control differences in the longwave UV-A(350–400 nm); however, the treatment/contro1 differences remained in the shortwave UV-A(320–350 nm), which was restricted by the glass, and in the total UV-A. The treatment/control differences persisted in the shortwave UV-A for the higher irradiance level, even under high summer ambient light. Also, spectral ratios (UVB:UV-A and shortwave: longwave UV-A) for all treatment groups decreased as the ambient UV-A radiation increased. Therefore, a range of experimental conditions exist where PE-covered lamps do not provide adequate control for UV-A irradiance, relative to the CA treatment, for glasshouse/growth chamber experiments. Potential complications in the interpretation of plant response exist for UV-B experiments conducted under low ambient light conditions (e.g. growth chambers; glasshouse in winter) or high daily UV-B irradiances (e.g. 14 kJ m^-2) for those plant responses that are sensitive to UV-A radiation.     

IDENTITY OF THE PHOTORECEPTORS FOR THE PERCEPTION OF IRRADIANCE IN PHOTOSYNTHETIC LIGHT ACCLIMATION IN TOMATO*

The photoreceptors involved in the photosynthetic acclimation of tomato (Lycopersicon esculentum Mill.) to increased irradiance were investigated. Plants were transferred from 100 p.mol m^?2 s^?1 cool white fluorescent light to higher irradiances of white light or white light supplemented with blue, red, green or yellow light. In these experiements light of all wavelengths tested was capable of causing acclimation as measured by the rate of light-saturated photosynthesis. It was concluded that the photosynthetic system rather than the blue-absorbing photoreceptor or phytochrome system acts as the photoreceptor for increased irradiance. No acclimation was observed in response to increased CO₂ levels, but increasing light integral at a constant irradiance was effective in bringing about acclimation. We conclude that acclimation is a response to increased photosynthetic light capture rather than increased photosynthetic carbon fixation, and involves a photon counting mechanism.     

Hydrochemistry of an ancient traditional irrigation reservoir in Padaviya,Sri Lanka

Padaviya Wewa Reservoir constructed in 2nd century BC is an important traditional reservoir in Sri Lanka. The construction and the use of these traditional Sri Lankan reservoirs are different from modern large-scale hydroelectric and multi-purpose reservoirs in Sri Lanka and the rest of the world. The hydrochemistry of these unique water bodies has been rarely studied and understood. In this study, depth-wise distribution of Fe, Mn, Zn, Cd, and As and their correlation with pH, dissolved oxygen, conductivity, temperature, BOD, COD, and total phosphate were investigated to identify the mechanisms that drive the water quality in Padaviya Wewa Reservoir. There is a well distinguishable chemical gradient from the surface to the bottom of the water column suggesting that Padaviya is not a well-mixed water body even though the maximum water depth during the study period was 4.75 m. There was a strong negative correlation between Fe, Mn, Zn, and Cd and DO in both surface and bottom waters. In addition, a strong positive correlation between Fe, Mn, Zn, Cd, and As, and total phosphate was observed in bottom waters. The suggested driving force behind the depth-wise distribution of chemical species is intense biological activities in the reservoir. The weak thermal stratification is strengthened by photosynthesis at the surface waters and organic matter degradation at bottom of the reservoir. The end-result of these biological activities are oxygen rich surface waters with low concentrations of nutrients and metal ions and oxygen depleted bottom waters with higher concentrations of nutrients and metal ions.     

Determination of organophosphorus pesticides by gas chromatography with mass spectrometry using a large‐volume injection technique after magnetic extraction

A fast and efficient method was developed for the extraction and determination of organophosphorus pesticides in water samples. Organophosphorus pesticides were extracted by solid‐phase extraction using magnetic multi‐walled carbon nanotubes and determined by gas chromatography with ion‐trap mass spectrometry. Parameters affecting the extraction were investigated. Under optimum conditions of the method, 10 mg magnetic multi‐walled carbon nanotubes were added into 10 mL sample. After 2 min, adsorbent particles settled at the bottom of test tube with a magnet. After removing aqueous supernatant, the analytes were desorbed with acetonitrile. Then, 70 μL of acetonitrile phase was injected into the gas chromatography and mass spectrometry system that had an ion‐trap analyzer. To achieve high sensitivity, the large‐volume‐injection technique was used with a programmed temperature vaporization inlet, and the ion‐trap mass spectrometer was operated in single ion storage mode. Under the best conditions, the enrichment factors and extraction recoveries were in the range of 113–124 and 74–103%, respectively. The limits of detection were between 3 and 15 ng/L, and the relative standard deviations were < 10%. This method was successfully used for the determination of organophosphorus pesticides in dam water, lagoon water, and river water samples with good reproducibility and recovery.     

Determination of Temperature Gradients with Micrometric Resolution by Local Open Circuit Potential Measurements at a Scanning Microelectrode

A method to determine localized temperature profiles using a scanning electrochemical microscopy (SECM) setup and potentiometry is presented. A Pt microelectrode was first calibrated to correlate the open circuit potential (OCP) with temperature in an electrolyte containing ferri/ferrocyanide. Using the calibration graph, the temperature at a given position and a time could be derived. For dynamic measurements, the thermal expansion of the surface was initially determined using shear force mode SECM. Following the OCP at the microelectrode static as well as dynamic temperature gradients above the heated surface were successfully probed and visualized with vertical micrometric resolution and with precision in temperature determination below 1 °C.     

A LABORATORY INVESTIGATION OF TWO ULTRAVIOLET SPECTRORADIOMETERS

Abstract— Spectral measurements of the solar UV irradiance at the earth's surface depend critically on the characteristics of the spectroradiometers and the procedures used in their calibration. Two UV spectroradiometers were subjected to a series of laboratory tests to investigate which factors were most significant in limiting the reliability of the absolute irradiance measurements. Three independent standards of spectral irradiance were scanned by both instruments under a range of bench conditions. The results were consistent to within about 3%, most of the uncertainty being due to scattered light in the laboratory. An UV laser was used to determine the slit function of each spectrometer and the influence of internal stray light. Significant departures from the ideal cosine and azimuth responses were measured by a xenon lamp. Both spectroradiometers were kept indoors throughout the experiments. The relevance of these laboratory results is discussed with respect to the task of measuring solar UV radiation in the field.     

The Distribution of Arsenic Compounds in the Ocean: Biological Activity in the Surface Zone and Removal Processes in the Deep Zone

The vertical profies of inorganic arsenic [As(III)+As(V)], monomethylarsonic acid (MMAA) and dimethylarsinic acid (DMAA) were investigated at four sampling stations in the Pacific Ocean and a sampling station in the southern Tasman Sea. In addition, the concentrations of those compounds in surface waters of the Pacific Ocean and Tasman Sea have been determined. The vertical profiles of inorganic arsenic showed the low concentrations in both the surface and deep/bottom zones. The depleted concentrations in the surface zone varied from 1000 to 1700 ng dm⁻³ and that in the deep/bottom zone varied from 1300 to 2050 ng dm⁻³. The maximum concentrations that varied from 1500 to 2450 ng dm⁻³ were usually observed at a depth of about 2000 m. Both MMAA and DMAA were observed throughout the water column at sampling stations in the north-western and equatorial regions of the Pacific Ocean. At the sampling station in the central northern Pacific gyre, DMAA was the only methylated arsenic compound observed throughout the water column. On the contrary, at the sampling station in the southern Tasman Sea, the only detected methylated arsenic compound throughout the water column was MMAA. Their vertical profiles showed maximum concentrations in the surface water which abruptly dropped with depth from 0 to 200 m. The concentration in the surface water was close to 10 ng dm⁻³ for MMAA and varied from 27 to 185 ng dm⁻³ for DMAA. At depths greater than 100 m, MMAA and DMAA were at comparable concentrations which varied from 0.7 to 14 ng dm⁻³. The low inorganic arsenic concentration in the surface zone was due to biological activity. This activity resulted in the uptake of As(V) and subsequent reduction and methylation to MMAA and DMAA. DMAA was the main predominant arsenic compound resulting from biological activity in surface waters. The low inorganic arsenic concentrations in the deep and bottom zones were likely to be caused by the adsorption of dissolved inorganic arsenic onto sinking particulates rich in iron and manganese oxides.     

Early phase laser induced plasma diagnostics and mass removal during single-pulse laser ablation of silicon

The electron number density and temperature during the early phase (<300 ns) of laser-induced plasmas from silicon using a 266-nm, 3-ns Nd:YAG laser were deduced via spectroscopic methods. These parameters were measured as a function of delay time vs. irradiance in the range of 2–80 GW/cm², and compared with crater volume measurements. A dramatic change in plasma characteristics (electron number density, temperature, and degree of ionization) as well as a sharp increase of mass removal was observed when the irradiance was increased beyond a threshold of 20 GW/cm². Possible mechanisms such as inverse bremsstrahlung and self-regulation were used to describe these data in the low irradiance region. Laser self-focusing and critical temperature are discussed to explain the dramatic changes after the irradiance reaches the threshold.     

THE ACTION SPECTRUM OF DAPHNIA MAGNA (CRUSTACEA) PHOTOTAXIS IN A SIMULATED NATURAL ENVIRONMENT

Abstract— The action spectrum of phototaxis in Daphnia magna (Crustacea) was measured in a chamber which simulated a natural angular distribution of underwater light. A 17% step-down in irradiance was used to stimulate the phototactic response at all wavelengths and irradiances tested. Peaks in the spectral response curves depended on the fluence rate to which the zooplankton were acclimated. The wavelength of maximum response (Z_max) shifted from yellow-green at the highest acclimation fluence rate (5.1 × 10⁻² Wm⁻²) to blue-violet at moderate rates. At low acclimation fluence rates, the blue-violet maximum was retained and another maximum developed in the red. At the lowest fluence rate (1.6 × 10⁻⁵ Wm⁻²), the blue-violet and red maxima were lost and another maximum developed in the near ultraviolet. The action spectrum indicates the presence of three, and possibly four, photopigments with Z_max, at ∼405, 440, 570 and 690nm. The 440 and 690nm maxima may belong to the same photopigment; however, this was not tested. Changes in zooplankton swimming speed, caused either by large changes in irradiance or by mechanical stimuli, were accompanied by changes in the strength of the phototactic response to the −17% stimulus at any irradiance level for white and monochromatic light, and indicated the presence of a mechanism connecting swimming speed and photosensitivity.     

THE STUDY OF ADAPTABILITY OF Drepanopus bispinosus TO TEMPERATURE AND SALINITY IN THE ANTARCTIC BURTON LAKE

The tolerance to temperautre and salinity of a Calanoida copepod: Drepanopus bispinosus from the Burton Lake in the Vestfold Hills, Antarctica, was tested in each season. The results obtained show that the copepod has a marked physiological property of tolerance to low temperature and high salinity, and this property could be strengthened after winter. The response to different temperature and salinity of the copepod evidently expressed the tardy characteristics contrasted with seasons. This means that for a long term in early winter(lower temperature and higher salinity) the animal still has higher tolerance to lower salinity and higher temperature; and it is opposite in early summer. These physiological features are possibly caused by natural acclimatization in the lake. The experimental data were processed and plotted by computer. The correlation of animal's survival rate with the changes of temperature and salinity presented an approximately smooth surface of a three-degree elliptic sphere.     

Stratification and tributyltin variability in San Diego Bay

Tributyltin (TBT), a biocidal antifoulant in many marine paints, was measured in near-surface and near-bottom water over a 25 h period at the entrance to a marina in San Diego Bay, USA. Surface water concentrations varied from 20 to 225 nanograms per liter (ng dm^?3) as TBT chloride and bottom water varied from non-detectable (<1 ng dm^?3) to 77 ng dm^?3. Surface water concentrations varied, with highest concentrations associated with ebbing tides, and lowest concentrations with flooding tides, indicating that the yacht basin is a source of TBT. Bottom water TBT concentrations were almost always lower than corresponding surface water concentrations. The highest bottom water concentrations were associated with flooding tides and lowest surface water TBT concentrations. Physical water column measurements indicate that vertical stratification developed during ebbing tides and dissipated during flooding tides. This accounted for maximum bottom water and minimum surface water TBT concentrations during flooding tides, due, at least in part, to vertical mixing and dilution during flood tides.     

Solar UVB and plant damage irradiances for different Argentinean regions

We calculated the integrated UVB and plant damage irradiances for Argentina, a country in the Southern Hemisphere spread over a large latitudinal range. The irradiances were calculated for clear sky days using the Madronich code for the average conditions of the months corresponding to the summer and winter solstices and the fall and spring equinoxes. Ozone, aerosol and ground albedo typical for each region and for each period of the year have been considered. A comparison was made of the behavior of these irradiances at the different locations. A more pronounced time dependence of the plant damage irradiance was obtained because of the fact that the corresponding spectrum is largely concentrated at a small wavelength of the UVB interval. We established a correlation between both irradiances, which can be approximated by a quadratic function. Because the plant damage irradiance is a quantity that is not directly measured by instruments, we showed the utility of the correlation by determining this biological effectiveness from the integrated UVB irradiance measured at the Astronomical Observatory of Rosario, Argentina, on clear sky days of the year 2001, as a characteristic example of the midlatitude near-sea level location of a highly productive agricultural region, which can be extended to other regions of the world. The plant damage results are relative ones (as is the case for the erythemal irradiance). So, they can be used to determine the maximum/minimum and asymmetry ratios, to study the influence of atmospheric variables and to make comparisons with other geographical locations.     

Ultraviolet B-photoprotection efficiency of mesocosm-enclosed natural phytoplankton communities from different latitudes: Rimouski (Canada) and Ubatuba (Brazil)

Photoprotection against UV-B radiation (UVBR; 280-320 nm) was examined in natural phytoplankton communities from two coastal environments at different latitudes: temperate Rimouski (Canada) and tropical Ubatuba (Brazil). Mesocosm experiments were performed at these sites to examine the response of phytoplankton to increases in UVBR that corresponded to local depletions of 30% and 60% in atmospheric ozone levels (low and high UVBR treatments, respectively). A fluorescence method using a pulse amplitude modulation fluorometer (Xe-PAM, Walz, Germany) with selective UV filters was used to estimate photoprotection, and these results were compared with an index of mycosporine-like amino acid (MAA) concentrations determined using spectrophotometry of methanol extracts. The present study provided the first evidence, to our knowledge, of the suitability of this in vivo fluorescence method for the estimation of UV photoprotection efficiency in natural phytoplankton. No significant differences were found for most of the variables analyzed between the light treatments used at both sites, but differences were found between sites throughout the duration of the experiments. Vertical mixing, used to maintain cells in suspension, likely alleviated serious UVBR-induced damage during both experiments by reducing the length of time of exposure to the highest UVBR irradiances at the surface. In Rimouski, this was the main factor minimizing the effects of treatment, because optical properties of the coastal seawater rapidly attenuated UVBR throughout the water column of the ca 2 m deep mesocosms. In this location, synthesis of MAAs and photoprotective pigments likely contributed to the observed phototolerance of phytoplankton and, hence, to their growth; however, in a comparison of the UVBR treatments, these variables showed no differences. In Ubatuba, where nutrient concentrations were significantly lower than those in Rimouski, light attenuation was less than that in Rimouski and UVBR reached the bottom of the mesocosms. UVBR penetration and the forced vertical mixing of the cells, without the possibility of vertical migration below this photostress zone, resulted in photo-inhibition, because confinement in the mesocosms forced cells to remain constantly exposed to high levels of irradiance during the daytime. Hence, additional effects of UVBR were masked in this experiment, because cells were damaged too much and phytoplankton populations were rapidly declining. There was also an overall preservation of MAAs, in contrast with chlorophyll (Chl) degradation, in spite of the fact that this UV screening was not sufficient to counteract photo-inhibition, which suggests an important role for these molecules, either in the overall photoprotection strategy or in other physiological processes. Altogether, local water characteristics, namely attenuation, mixing, and nutrients concentration, can strongly modulate the photoprotection strategies used by natural phytoplankton populations in coastal environments.     

Multisyringe chromatography (MSC) using a monolithic column for the determination of sulphonated azo dyes

A methodology based on multisyringe chromatography with a monolithic column was developed to determine three sulphonated azo textile dyes: Acid Yellow 23, Acid Yellow 9 and Acid Red 97. An ion pair reagent was needed because of the low affinity between the monolithic column and the anionic dyes. The proposed analytical system is simple, versatile and low-cost and has great flexibility in manifold configuration.The method was optimized through experimentation based on experimental design methodology. For this purpose two blocks of full factorial 2³ were done sequentially. In the first experimental plan, the factors studied were: the % of acetonitrile in organic phase, the % of H₂O in the mobile phase and the kind of ion pair reagent. In this stage, a simple configuration was used which has only one syringe for the mobile phase.After the first experimentation, we added a second syringe with a second mobile phase to the multisyringe module and performed a second full factorial 2³. The factors studied in this case were: the % of acetonitrile in the second mobile phase, the pH and the concentration of ion pair reagent in both mobile phases. After this design, the optimal conditions were selected for obtaining a good resolution between the peaks of yellow dyes (1.47) and the elution of red dye in less than 8 min.The methodology was validated by spiking different amounts of each dye in real water samples, specifically, tap water, well water and water from a biological wastewater lagoon.