Simplifying the Centrolene buckleyi complex (Amphibia: Anura: Centrolenidae): a taxonomic review and description of two new species.

Centrolenidae is a Neotropical family widely distributed in Central and South America, with its species richness concentrated in the tropical Andes. Several taxonomic problems have been identified within this family, mostly related to species with broad geographic distributions. In this study, we assessed and redefined the species boundaries of the Centrolene buckleyi species complex, and formally described two new species from the Andes of Ecuador. These new taxa are recognized by a combination of morphometric, osteological, acoustic, and genetic data. Following IUCN criteria, we propose that the two new species should to be considered as Endangered (EN), mainly because of their small distributions and habitat loss. The C. buckleyi complex provides insights into the biogeography of closely related Andean species. As in other glassfrogs, speciation in Centrolene seems to be mediated by the linearity of the Andes, where gene flow can be restricted by topography and, also, local extinctions.

Bioactivity of synthetic peptides from Ecuadorian frog skin secretions against Leishmania mexicana, Plasmodium falciparum, and Trypanosoma cruzi.

Chagas disease, leishmaniasis, and malaria are major parasitic diseases disproportionately affecting the underprivileged population in developing nations. Finding new, alternative anti-parasitic compounds to treat these diseases is crucial because of the limited number of options currently available, the side effects they cause, the need for long treatment courses, and the emergence of drug-resistant parasites. Anti-microbial peptides (AMPs) derived from amphibian skin secretions are small bioactive molecules capable of lysing the cell membrane of pathogens while having low toxicity against human cells. Here, we report the anti-parasitic activity of five AMPs derived from skin secretions of three Ecuadorian frogs: cruzioseptin-1, cruzioseptin-4 (CZS-4), and cruzioseptin-16 from Cruziohyla calcarifer; dermaseptin-SP2 from Agalychnis spurrelli; and pictuseptin-1 from Boana picturata. These five AMPs were chemically synthesized. Initially, the hemolytic activity of CZS-4 and its minimal inhibitory concentration against Escherichia coli, Staphylococcus aureus, and Candida albicans were determined. Subsequently, the cytotoxicity of the synthetic AMPs against mammalian cells and their anti-parasitic activity against Leishmania mexicana promastigotes, erythrocytic stages of Plasmodium falciparum and mammalian stages of Trypanosoma cruzi were evaluated in vitro. The five AMPs displayed activity against the pathogens studied, with different levels of cytotoxicity against mammalian cells. In silico molecular docking analysis suggests this bioactivity may occur via pore formation in the plasma membrane, resulting in microbial lysis. CZS-4 displayed anti-bacterial, anti-fungal, and anti-parasitic activities with low cytotoxicity against mammalian cells. Further studies about this promising AMP are required to gain a better understanding of its activity.IMPORTANCEChagas disease, malaria, and leishmaniasis are major tropical diseases that cause extensive morbidity and mortality, for which available treatment options are unsatisfactory because of limited efficacy and side effects. Frog skin secretions contain molecules with anti-microbial properties known as anti-microbial peptides. We synthesized five peptides derived from the skin secretions of different species of tropical frogs and tested them against cultures of the causative agents of these three diseases, parasites known as Trypanosoma cruzi, Plasmodium falciparum, and Leishmania mexicana. All the different synthetic peptides studied showed activity against one of more of the parasites. Peptide cruzioseptin-4 is of special interest since it displayed intense activity against parasites while being innocuous against cultured mammalian cells, which indicates it does not simply hold general toxic properties; rather, its activity is specific against the parasites.

Path for recovery: an ecological overview of the Jambato Harlequin Toad (Bufonidae: Atelopus ignescens) in its last known locality, Angamarca Valley, Ecuador.

The Jambato Harlequin toad (Atelopus ignescens), a formerly abundant species in the Andes of Ecuador, faced a dramatic population decline in the 1980s, with its last recorded sighting in 1988. The species was considered Extinct by the IUCN until 2016, when a fortuitous discovery of one Jambato by a local boy reignited hope. In this study, we present findings from an investigation conducted in the Angamarca parish, focusing on distribution, abundance, habitat preferences, ecology, disease susceptibility, and dietary habits of the species. In one year we identified 71 individuals at different stages of development in various habitats, with a significant presence in agricultural mosaic areas and locations near water sources used for crop irrigation, demonstrating the persistence of the species in a complex landscape, with considerable human intervention. The dietary analysis based on fecal samples indicated a diverse prey selection, primarily comprising arthropods such as Acari, Coleoptera, and ants. Amphibian declines have been associated with diseases and climate change; notably, our study confirmed the presence of the pathogen Batrachochytrium dendrobatidis (Bd), but, surprisingly, none of the infected Jambatos displayed visible signs of illness. When analyzing climatic patterns, we found that there are climatic differences between historical localities and Angamarca; the temporal analysis also exposes a generalized warming trend. Finally, in collaboration with the local community, we developed a series of management recommendations for terrestrial and aquatic environments occupied by the Jambato.

A lifestyle adjustments program in long COVID-19 improves symptomatic severity and quality of life. A randomized control trial.

OBJECTIVE: To evaluate the efficacy of a therapeutic intervention based on self-adjustment strategies for improving of symptomatic severity and quality of life. METHODS: The study was a randomised single-blind clinical trial. Quality of life, disability, and functional impairment were collected. The control group received a leaflet with information on the main symptoms of Long-COVID-19 syndrome, in addition to standard medical treatment. The intervention group received treatment following a dual approach; on the one hand, monitoring and recognition of symptomatology and on the other hand, adaptation and functional improvement. RESULTS: A total of 54 participants were included, 27 were included in the intervention group and 27 in the control group. At the beginning of the study, no significant differences were found between groups. After intervention, the quality of life variable showed significant differences between groups in the self-care and anxiety/depression dimensions. Significant between-groups differences were also found for the self-care subscale of the disability variable. The intervention group showed significant differences from baseline on some subscales of the quality of life, disability, and functional impairment variables. CONCLUSION: Strategies based on lifestyle adjustments are adequate for the improvement of quality of life and symptom severity in the long COVID-19 population. PRACTICE IMPLICATION: The findings suggest that applying an intervention focused in self-adjustment for long COVID patients can have positive effects.

A case report of CBD and THC as analgesic therapy in a cat with chronic osteoarthritic pain.

A 10-year-old mixed breed male cat presented with clinical signs related to chronic orthopaedic pain. Upon physical examination, pain was noted, based on the feline Musculoskeletal Pain Index (FMPI). An analgesic treatment with a full spectrum cannabis oil (1.8% CBD and 0.8% THC) was proposed for 30 days (0,5 mg/kg based on CBD). The FMPI scale score decreased more than 50%. This case reported a satisfactory outcome for the patient and the owner, although this medication could increase ALT. Given the paucity of literature published to date on the treatment of veterinary species with cannabis-based medications, further clinical and pharmacokinetic studies are necessary to study the safety and efficacy of its use.

Random noise stimulation in the treatment of patients with neurological disorders.

Random noise stimulation technique involves applying any form of energy (for instance, light, mechanical, electrical, sound) with unpredictable intensities through time to the brain or sensory receptors to enhance sensory, motor, or cognitive functions. Random noise stimulation initially employed mechanical noise in auditory and cutaneous stimuli, but electrical energies applied to the brain or the skin are becoming more frequent, with a series of clinical applications. Indeed, recent evidence shows that transcranial random noise stimulation can increase corticospinal excitability, improve cognitive/motor performance, and produce beneficial aftereffects at the behavioral and psychological levels. Here, we present a narrative review about the potential uses of random noise stimulation to treat neurological disorders, including attention deficit hyperactivity disorder, schizophrenia, amblyopia, myopia, tinnitus, multiple sclerosis, post-stroke, vestibular-postural disorders, and sensitivity loss. Many of the reviewed studies reveal that the optimal way to deliver random noise stimulation-based therapies is with the concomitant use of neurological and neuropsychological assessments to validate the beneficial aftereffects. In addition, we highlight the requirement of more randomized controlled trials and more physiological studies of random noise stimulation to discover another optimal way to perform the random noise stimulation interventions.

Glacier fluctuations in the northern Patagonian Andes (44°S) imply wind-modulated interhemispheric in-phase climate shifts during Termination 1.

The Last Glacial Termination (T1) featured major changes in global circulation systems that led to a shift from glacial to interglacial climate. While polar ice cores attest to an antiphased thermal pattern at millennial timescales, recent well-dated moraine records from both hemispheres suggest in-phase fluctuations in glaciers through T1, which is inconsistent with the bipolar see-saw paradigm. Here, we present a glacier chronology based on 30 new 10Be surface exposure ages from well-preserved moraines in the Lago Palena/General Vintter basin in northern Patagonia (~ 44°S). We find that the main glacier lobe underwent profound retreat after 19.7 ± 0.7 ka. This recessional trend led to the individualization of the Cerro Riñón glacier by ~ 16.3 ka, which underwent minor readvances at 15.9 ± 0.5 ka during Heinrich Stadial 1, during the Antarctic Cold Reversal with successive maxima at 13.5 ± 0.4, 13.1 ± 0.4, and 13.1 ± 0.5 ka, and a minor culmination at 12.5 ± 0.4 ka during Younger Dryas time. We conclude that fluctuations of Patagonian glaciers during T1 were controlled primarily by climate anomalies brought by shifts in the Southern Westerly Winds (SWW) locus. We posit that the global covariation of mountain glaciers during T1 was linked to variations in atmospheric CO2 (atmCO2) promoted by the interplay of the SWW-Southern Ocean system at millennial timescales.

Laser Prophylaxis in Patients with Stickler Syndrome.

PURPOSE: To evaluate the association among laser prophylaxis treatment, retinal detachment (RD), and visual acuity (VA) in patients with Stickler syndrome (SS). DESIGN: Retrospective comparative case series. PARTICIPANTS: Patients with SS. METHODS: Patients received extended vitreous base laser (EVBL), nonprotocol laser (NPL), or no laser prophylaxis treatment of any kind. MAIN OUTCOME MEASURES: The 2 main outcome measures that were examined in these patients were rates of RD and VA. RESULTS: In this study, 230 eyes of 115 patients were included. Fifty-nine patients were women (51%). The median age at the time of laser prophylaxis treatment was 9.5 years (interquartile range [IQR], 6-13 years), and the median age of patients with RD was 11 years (IQR, 7-18 years). Of the 230 eyes, 92 did not undergo any laser treatment, 9 received NPL treatment, and 129 received EVBL treatment. Of the 129 eyes that underwent EVBL treatment, 4 (3%) had RD, compared with 74 eyes (73%) that had RD and did not receive laser or NPL treatment (P < 0.001). Eyes that received EVBL treatment had approximately 8 lines better vision, on average, compared with those that did not receive laser or NPL treatment (-0.86 logarithm of the minimum angle of resolution; 95% confidence interval,-1.1 to -0.64; P < 0.001). CONCLUSIONS: Treatment with EVBL seems to reduce the rate of subsequent RD and is associated with better VA in patients with SS.

Safety and clinical effectiveness of intravitreal administration of bevacizumab (Lumiere®) in patients with neovascular age-related macular degeneration.

The present study was an open-label, prospective, uncontrolled and multicenter clinical trial to investigate the safety and effectiveness of bevacizumab (Lumiere®) administered by the intravitreal route for the treatment of neovascular age-related macular degeneration (nAMD). A total of 22 patients without previous treatment with anti-vascular endothelial growth factor were recruited. Monthly therapy with 1.25 mg intravitreal bevacizumab was applied. Adverse events (AE), visual acuity (VA) and central retinal thickness (CRT) were assessed at baseline, day 1 and day 28 after each injection. A total of 87 AEs were reported; most of them were not serious (96.6%), expected (65.5%) and occurred after the third injection (56.3%). The most frequent AE was 'conjunctival hemorrhage' (29.9% of AEs), attributed to the injection procedure. Treatment was not suspended due to safety reasons in any case. After six months, a statistically significant gain of +8.2 (SD±8.8) letters and a CRT reduction of -75.50 µm (SD±120.3) were achieved with unilateral therapy. VA improvement and CRT reduction were also achieved with bilateral therapy, although to a lesser extent. The results of the present study suggested that therapy with a minimum of 3 doses of bevacizumab over a 6-month period was well tolerated and resulted in a sustained response regarding VA improvement and CRT reduction from the beginning of therapy compared with the baseline value. The study protocol was registered at clinicaltrials.gov (ref. no. NCT03668054).

Prevalencia de embarazos con infección por virus Zika en una Unidad de Primer Nivel de Atención en Coatzacoalcos Veracruz México del Instituto Mexicano del Seguro Social / PREVALENCE OF PREGNANCIES WITH ZIKA VIRUS INFECTION IN A FIRST LEVEL CARE UNIT IN COATZACOALCOS VERACRUZ MEXICO OF THE MEXICAN INSTITUTE OF SOCIAL SECURITY

El virus Zika, es un arbovirus del género flavivirus (familia Flaviviridae), muy cercano filogenéticamente al del dengue, fiebre amarilla, la encefalitis japonesa, o el virus del Nilo Occidental. El presente estudio exploratorio busca identificar los casos confirmados y sugestivos de infección por virus Zika en mujeres embarazadas en la Unidad de Medicina Familiar N° 60 del IMSS de Coatzacoalcos, Veracruz. México como primera aproximación epidemiológica en esta región.

The Zika virus is an arbovirus of the genus flavivirus (family Flaviviridae), very closely phylogenetically a virus such as dengue fever, yellow fever, Japanese encephalitis, or West Nile virus. According to the Epidemiological Surveillance System for Zíka Virus Disease, until November 4th, 2016, in Mexico there were 6,094 confirmed cases of infection and 3,167 suspected cases of Zika virus infection in pregnant women in 24 states of the Mexican Republic, with a higher prevalence in Veracruz, Chiapas, Colima, Campeche, Yucatan and Quintana Roo. This is an original study that focuses on identifying the confirmed and suggestive cases of Zika virus infection in pregnant women in the Family Medicine Unit No. 60 of the IMSS in Coatzacoalcos, Veracruz. Mexico; It is an exploratory study in the registry of cases of pregnant women with Zika virus infection in the Family Medicine Unit No. 60 of Coatzacoalcos, Veracruz, Mexico as epidemiological basis

Seguimiento de hijos de madres con infección por virus ZIKA en una Unidad de Primer Nivel de Atención Médica de Coatzacoalcos Veracruz, México del Instituto Mexicano del Seguro Social / Follow-Up of congenital Zika virus Infection

El virus Zika, es un arbovirus del género flavivirus (familia Flaviviridae), muy cercano filogenéticamente a virus como el dengue, fiebre amarilla, la encefalitis japonesa, o el virus del Nilo Occidental. El presente estudio es de tipo exploratorio con base en el registro de casos de recién nacidos y lactantes hijos de mujeres embarazadas con infección de virus Zika en la Unidad de Medicina Familiar Nº 60 de Coatzacoalcos, Veracruz, dentro del período de diciembre del 2016 a julio del 2017. Se realizó un estudio polietápico que constó de revisión de casos por medio de uso de expediente clínico en la Consulta Externa en la Unidad Médico Familiar Nº 60 del IMSS de Coatzacoalcos, Veracruz. como primera aproximación epidemiológica en esta región en recién nacidos y lactantes hijos de madres con infección por virus Zika, que posteriormente fueron sometidos a estudios de valoración oftalmológica, neurológica y somatométrica; con ayuda del servicio de Salud Pública de dicha unidad (AU)

Zika virus is an arbovirus of the genusflavivirus (family Flaviviridae), very close phylogenetically to viruses such as denque, yellow fever, japanese encephalitis, or West Nile virus. The present study is exploratory based on the registry of cases of newborns and infants born to pregnant women with zika virus infection in the Family Medicine Unit Nº 60 of Coatzacalcos, Veracruz, within the period of December 2016 to July 2017. A multistage study was carried out that consisted of the review of cases through the use of a clinical file in the Outpatient Consultation in the Family Medical Unit Nº 60 of the IMSS. of Coatzacoalcos, Veracruz. As a first epidemiological approach in this region in newborns and infants born to mothers with Zika virus infection, who were subsequently subjected to ophtalmological, neurological and somatometric assessment studies; with the help of the Public Health service of said unit (AU)

Next generation flow for minimally-invasive blood characterization of MGUS and multiple myeloma at diagnosis based on circulating tumor plasma cells (CTPC).

Here, we investigated for the first time the frequency and number of circulating tumor plasma cells (CTPC) in peripheral blood (PB) of newly diagnosed patients with localized and systemic plasma cell neoplasms (PCN) using next-generation flow cytometry (NGF) and correlated our findings with the distinct diagnostic and prognostic categories of the disease. Overall, 508 samples from 264 newly diagnosed PCN patients, were studied. CTPC were detected in PB of all active multiple myeloma (MM; 100%), and smoldering MM (SMM) patients (100%), and in more than half (59%) monoclonal gammopathy of undetermined significance (MGUS) cases (p <0.0001); in contrast, CTPC were present in a small fraction of solitary plasmacytoma patients (18%). Higher numbers of CTPC in PB were associated with higher levels of BM infiltration and more adverse prognostic features, together with shorter time to progression from MGUS to MM (p <0.0001) and a shorter survival in MM patients with active disease requiring treatment (p ≤ 0.03). In summary, the presence of CTPC in PB as assessed by NGF at diagnosis, emerges as a hallmark of disseminated PCN, higher numbers of PB CTPC being strongly associated with a malignant disease behavior and a poorer outcome of both MGUS and MM.

[Laparoscopic repair of inguinal hernia and communicating hydrocele in children. Our experience in the last years]. / Reparación laparoscópica de la hernia inguinal e hidrocele en edad pediátrica. Nuestra experiencia en los últimos años.

INTRODUCTION: Laparoscopic inguinal hernia and hydrocele repair has become an alternative to the more frequently used inguinal approach. We describe our experience with the laparoscopic technique. MATERIAL AND METHODS: Retrospective study of laparoscopic inguinal hernia and hydrocele repair performed during the period between 2003-2016. We included patients < 14 years, with communicating hydrocele and/or indirect inguinal hernia diagnosis (N = 39). We have studied two cohorts; group A patients with indirect inguinal hernia (N = 31) and group B patients with communicating hydrocele (N = 8). We used intraperitoneal laparoscopic approach, performing herniorrhaphy of the internal inguinal orifice without resection of the hernial sac. The main outcome measurements were surgical indication and postoperative complications. RESULTS: The indication for laparoscopic repair was bilateral defects (46.2%), unilateral defects (28.2%), defects associated with umbilical hernia (23.1%) and associated with another pathology (2.6%). In group A the rate of complications that required new admission was 3.2% and the recurrence rate was 9.7%. In group B, was 12.5% ​​and 12.5% respectively. We have not observed a significant relationship with respect to patient age, laterality or suture used and the rate of complications in any of the groups. CONCLUSIONS: Despite the limitations of our study given the small sample size and retrospective nature, the complication rate (15%) is higher than that published in the literature. This makes us take a step back to reassess our indications and technique, and not forget that the laparoscopic approach is not a technique without risks and requires a learning curve.

OBJETIVOS: La reparación laparoscópica de la hernia inguinal e hidroceles en la edad pediátrica, se ha convertido en una alternativa a la herniorrafia inguinal clásica. Describimos nuestra experiencia en las intervenciones realizadas con dicha técnica. MATERIAL Y METODOS: Estudio retrospectivo de las reparaciones laparoscópicas de hernia inguinal e hidrocele realizadas en el periodo 2003-2016. Se han incluido pacientes <14 años, diagnosticados de hidrocele comunicante y/o de hernia inguinal indirecta (N=39), divididos en dos cohortes: grupo A pacientes con hernia inguinal indirecta y grupo B pacientes con hidrocele comunicante. Se ha empleado el abordaje laparoscópico intraperitoneal, y anillorrafia del orificio inguinal interno sin resección del saco herniario. Las variables principales estudiadas han sido la indicación quirúrgica y las complicaciones postquirúrgicas. RESULTADOS: Las indicaciones quirúrgicas han sido los defectos bilaterales (46,2%), defectos unilaterales (28,2%), defectos asociados a hernia umbilical (23,1%) y asociados a otra patología (2,6%). En el grupo A la tasa de complicaciones que precisaron de ingreso fue de 3,2% y la tasa de recurrencias fue de 9,7%; mientras que en el B han sido del 12,5% y del 12,5% respectivamente. No hay una relación significativa respecto a la edad, la lateralidad o la sutura utilizada y la tasa de complicaciones en ninguno de los grupos. CONCLUSIONES: A pesar de las limitaciones del estudio, el porcentaje total de complicaciones (15,3%) es más elevado que el publicado en la literatura. Esto nos hace reflexionar y no olvidar que el abordaje laparoscópico no es una técnica exenta de riesgos y requiere de una curva de aprendizaje.

Stable symmetry-protected 3D embedded solitons in Bose-Einstein condensates.

Embedded solitons are rare self-localized nonlinear structures that, counterintuitively, survive inside a continuous background of resonant states. While this topic has been widely studied in nonlinear optics, it has received almost no attention in the field of Bose-Einstein condensation. In this work, we consider experimentally realizable Bose-Einstein condensates loaded in one-dimensional optical lattices and demonstrate that they support continuous families of stable three-dimensional (3D) embedded solitons. These solitons can exist inside the resonant continuous Bloch bands because they are protected by symmetry. The analysis of the Bogoliubov excitation spectrum as well as the long-term evolution after random perturbations proves the robustness of these nonlinear structures against any weak perturbation. This may open up a way for the experimental realization of stable 3D matter-wave embedded solitons as well as for monitoring the gap-soliton to embedded-soliton transition.

Next Generation Flow for highly sensitive and standardized detection of minimal residual disease in multiple myeloma.

Flow cytometry has become a highly valuable method to monitor minimal residual disease (MRD) and evaluate the depth of complete response (CR) in bone marrow (BM) of multiple myeloma (MM) after therapy. However, current flow-MRD has lower sensitivity than molecular methods and lacks standardization. Here we report on a novel next generation flow (NGF) approach for highly sensitive and standardized MRD detection in MM. An optimized 2-tube 8-color antibody panel was constructed in five cycles of design-evaluation-redesign. In addition, a bulk-lysis procedure was established for acquisition of ⩾107 cells/sample, and novel software tools were constructed for automatic plasma cell gating. Multicenter evaluation of 110 follow-up BM from MM patients in very good partial response (VGPR) or CR showed a higher sensitivity for NGF-MRD vs conventional 8-color flow-MRD -MRD-positive rate of 47 vs 34% (P=0.003)-. Thus, 25% of patients classified as MRD-negative by conventional 8-color flow were MRD-positive by NGF, translating into a significantly longer progression-free survival for MRD-negative vs MRD-positive CR patients by NGF (75% progression-free survival not reached vs 7 months; P=0.02). This study establishes EuroFlow-based NGF as a highly sensitive, fully standardized approach for MRD detection in MM which overcomes the major limitations of conventional flow-MRD methods and is ready for implementation in routine diagnostics.

[Total reverse shoulder replacement. Evaluation of the clinical results and complications in a series of 52 cases]. / Artroplastia total invertida de hombro. Evaluación de resultados clínicos y complicaciones tras una serie de 52 casos.

OBJECTIVE: To evaluate the clinical results and analyse the complications of total reverse shoulder replacement performed in our centre over an 8 year period. MATERIAL AND METHOD: A retrospective study was conducted on 50 patients (52 shoulders), with a mean age of 70.15 years (range 51 to 84 years) between December 2004 and December 2012, who received a total reverse shoulder replacement, all performed by the same surgeon. The results have been evaluated according to clinical data, radiography study, a satisfaction scale, and the Constant scale, with a minimum follow-up of 16 months. Five of the cases (9.62%) had been intervened due to fractures of the proximal end of the humerus, 6 cases (11.53%) as surgical consequence of a prosthesis revision, 10 cases (19.23%) due to fracture sequelae, and 30 cases (59.62%) were patients with arthropathy due to a massive fracture of the rotator cuff. RESULTS: After a mean follow up of 35.78 months (range, 16-82), satisfactory clinical results were obtained in 80% of cases, with a mean preoperative Constant of 27.7 points, and reaching 67.1 points 12 months after the operation. On the visual analogue scale, 8.25 points were obtained before the surgery, which decreased to 2.25 points 12 months later. The complications rate was 15.38%, which were due to an intra-operative fracture (1.92%), deep infection (3.84%), instability (3.84%), and early mechanical loosening (3.84%). Scapular notching was observed in the radiographic study in 9 (17.3%) cases. DISCUSSION AND CONCLUSIONS: After the results obtained, it could be said that total reverse shoulder replacement achieved encouraging results in the short term for the treatment of glenohumeral arthrosis and massive tears of the rotary cuff. On analysing our series, it can be seen that the complications rate is much higher when it is used to treat fracture sequelae in which there is a loss of proximal humerus bone stock.

A simple and sensitive vortex assisted liquid-liquid microextraction method for the simultaneous determination of haloanisoles and halophenols in wines.

A vortex assisted liquid-liquid microextraction (VALLME) method was developed and optimised for the determination of the main compounds that can cause cork taint in wines, 2,4,6-trichloroanisole (TCA), 2,3,4,6-tetrachloroanisole (TeCA), 2,4,6-tribromoanisole (TBA) and pentachloroanisole (PCA); and their corresponding halophenolic precursors. Target compounds were determined by gas chromatography combined with a micro-electron capture detector (GC-µECD) system. Halophenol extraction and derivatisation processes were performed at the same time. To optimise the VALLME method, the extraction solvent was selected. Then, the other parameters of influence, such as volume of extraction solvent and derivatisation agent, salt addition and vortex time were optimised using a central composite design combined with desirability functions. Once the optimal conditions had been determined, the method was validated, showing satisfactory linearity (with correlation coefficients over 0.983), repeatability (below 10.0%) and reproducibility (below 11.2%). Detection limits obtained were lower than the olfactory threshold of the studied compounds, being similar or even lower than previously reported with the advantage of reducing the extraction time. The analysis of real wine samples demonstrated the applicability of the method. To our knowledge, this is the first time that VALLME has been applied for the simultaneous determination of haloanisoles and halophenols in wine.