Simultaneous Determination of Six Residual Solvents in Phenylbutazone Raw Material by Head-space GC / 中国药房

OBJECTIVE： To establish a method for simultaneous determination of residual solvents in phenylbutazone raw material. METHODS： Head-space GC was adopted. The determination was performed on Agilent HP-5 capillary column by temperature programming. The temperature of injector was 200 ℃， and detector was flame ionization detector with temperature of 250 ℃； carrier gas was nitrogen （purity：99.99％） at the flow rate of 2.0 mL/min. The split ratio was 5 ∶ 1. Headspace equilibrium temperature was 60 ℃， and equilibration time was 30 min. The sample size was 1 mL. RESULTS： The linear range was 0.15-4.5   μg/mL for methanol （r＝0.999 9）， 0.25-7.5 μg/mL for ethanol （r＝0.999 7）， 0.25-7.5 μg/mL for isopropyl alcohol （r＝0.999 7）， 0.03-0.9 μg/mL for dichloromethane （r＝0.999 3）， 0.25-7.5 μg/mL for ethyl acetate （r＝0.999 3）， 0.044-1.32 μg/mL for N，N-dimethyl formamide （r＝0.999 3）， respectively. The limits of detection were 0.05， 0.08， 0.08， 0.01， 0.08， 0.015 μg/mL. The limits of quantitation were 0.15， 0.25， 0.25， 0.03， 0.25， 0.044 μg/mL. RSDs of precision test were lower than 2.0％. RSDs of stability and reproducibility tests were lower than 3.0％. The recoveries were 98.75％-100.12％ （RSD＝0.56％， n＝9）， 98.07％-101.20％ （RSD＝1.12％， n＝9）， 98.36％-100.80％ （RSD＝0.92％， n＝9）， 98.33％-101.67％ （RSD＝0.98％， n＝9）， 98.11％-100.40％ （RSD＝0.72％， n＝9） and 98.75％-101.05％ （RSD＝0.89％， n＝9）. CONCLUSIONS： The method is simple， accurate， precise， stable， reproducible and durable， and can be used for simultaneous determination of 6 residual solvents in phenylbutazone raw material.

Enhancing the dissolution of phenylbutazone using Syloid? based mesoporous silicas for oral equine applications / 药物分析学报

Three mesoporous silica excipients (Syloid? silicas AL-1 FP, XDP 3050 and XDP 3150) were formulated with a model drug known for its poor aqueous solubility, namely phenylbutazone, in an attempt to enhance the extent and rate of drug dissolution. Although other forms of mesoporous silica have been investigated in previous studies, the effect of inclusion with these specific Syloid? silica based excipients and more interestingly, with phenylbutazone, is unknown. This work reports a significant enhancement for both the extent and rate of drug release for all three forms of Syloid? silica at a 1:1 drug:silica ratio over a period of 30 min. An explanation for this increase was determined to be conversion to the amorphous form and an enhanced drug loading ability within the pores. Differences between the release profiles of the three silicas were concluded to be a consequence of the physicochemical differences between the three forms. Overall, this study confirms that Syloid? silica based excipients can be used to enhance dissolution, and potentially therefore bioavailability, for compounds with poor aqueous solubility such as phenylbutazone. In addition, it has been confirmed that drug release can be carefully tailored based on the choice of Syloid? silica and desired release profile.

Efficiency of transabdominal ultrasound examination in aid to early diagnosis of induced right dorsal colitis in horses / Eficiência do exame ultrassonográfico transabdominal no auxílio ao diagnóstico precoce da colite dorsal direita induzida em equinos / Eficiencia del examen ecográfico transabdominal como ayuda en el diagnóstico precoz de la colitis dorsal derecha inducida en equinos

Abstract The Right Dorsal Colitis (CDD) is an enteropathy characterized by wall inflammation and edema and ulceration of the mucosa of the right dorsal colon in horses. Its occurrence is associated with prolonged use and/or overdoses of non-steroidal anti-inflammatory drugs (NSAIDs), especially phenylbutazone (FBTZ). Clinical abnormalities include anorexia, colic, hypoproteinemia, neutropenia, endotoxemia, diarrhea and weight loss. The ultrasound examination (EUS) is important in the diagnosis of abdominal abnormalities. Aiming to evaluate the efficiency of EUS in the early diagnosis of CDD, EUS were performed regularly in five horses (C1-C5) underwent protocol of induction experimental colitis. Five images per animal were taken on alternate days (D0-D14), between the 11th and 15th intercostal spaces in right side and, in each image, the wall thickness of the right dorsal colon was measured in millimeters (mm) at four different sites. There was no difference between D0 (3.4±0.55mm) and D3 (4.9 ±1.79 mm), but in D5 (7.17 ±1.28 mm), D7 (7.00 ±1.68 mm) and D9 (6.71 ±2.27 mm), there was significant increase in colon wall thickness in relation to D0 and D3 (p=0.05). Since the onset of relevant clinical signs, such as diarrhea was evident in all animals only in D9, it was concluded that ultrasonography of the right dorsal colon is sensitive method as early diagnosis of CDD.

A Colite Dorsal Direita (CDD) é uma enteropatia caracterizada por inflamação, edema mural e ulcerações da mucosa do cólon dorsal direito de equinos. Sua ocorrência associa-se ao uso prolongado e/ou sobredoses de antiinflamatórios n ão esteroidais (AINEs), principalmente fenilbutazona (FBTZ). As alterações clínicas incluem anorexia, cólica, hipoproteinemia, neutropenia, endotoxemia, diarreia e perda de peso. O exame ultrassonográfico (EUS) é importante como auxílio no diagnóstico de anormalidades abdominais. Com o objetivo de avaliar a eficiência do EUS no diagnóstico precoce da CDD foram realizados EUS periódicos em cinco equinos (C1-C5) submetidos a protocolo experimental de colites. Cinco imagens ultrassonográficas por animal foram obtidas em dias alternados (D0-D14), entre o 11º e 15º espaços intercostais direitos e, em cada imagem, a espessura da parede do cólon dorsal direito foi mensurada em milímetros (mm) em quatro áreas diferentes. N ão houve diferença entre D0 (3,4 ±0,55 mm) e D3 (4,9 ±1,79 mm), porém em D5 (7,17 ±1,28 mm), D7 (7,00 ±1,68 mm) e D9 (6,71 ±2,27 mm), constatou-se aumento significativo da espessura da parede do cólon em relação a D0 e D3 (p=0,05). Já que o início dos sinais clínicos importantes foi evidente nos animais somente em D9, concluiu-se que o EUS do cólon dorsal direito é sensível como método diagnóstico precoce da CDD.

La Colitis Dorsal Derecha (CDD) es una enteropatía caracterizada por inflamación, edema de pared y ulceraciones de la mucosa del colon dorsal derecho de equinos. Su ocurrencia se asocia al uso prolongado o sobredosis de antinflamatorios no esteroides (Aines), principalmente la fenilbutazona (FBTZ). Las alteraciones clínicas incluyen anorexia, cólico, hipoproteinemia, neutropenia, endotoxemia, diarrea y pérdida de peso. El examen ecográfico (EET) es importante como auxilio diagnóstico de anormalidades abdominales. Con el objetivo de evaluar la eficiencia del EET en el diagnóstico precoz de CDD fueron realizados EET periódicos en cinco equinos (C1- C5) sometidos a un protocolo experimental de colitis. Cinco imágenes ecográficas por animal fueron obtenidas en días alternados (D0-D14); entre el 11º y 15º espacios intercostales derechos y en cada imagen, el espesor del colon dorsal derecho fue medido en milímetros (mm) en cuatro áreas diferentes. No hubo diferencia entre D0 (3,4 ±0,55 mm) y D3 (4,9 ±1,79 mm), sin embargo en D5 (7,17 ±1,28 mm), D7 (7,00 ±1,68 mm) y D9 (6,71 ±2,27 mm), se estableció un aumento significativo del espesor de la pared del colon en relación a D0 y D3 (p=0,05). Debido al inicio evidente de signos clínicos importantes solo a partir de D9, se concluye que el EET del colon dorsal derecho es sensible como método diagnóstico precoz de CDD.

A study of anti-inflammatory activity of the benzofuran compound (3,4-dihydro 4-oxo-benzofuro [3,2-d] pyrimidine-2-propionic acid) in chronic model of inflammation.

Background: Benzofuran compounds are shown to have pharmacological properties such as antiarrhythmic, antidepressant, antifungal, and antibacterial activity. Some studies conducted on them have revealed that they are having anti-inflammatory property also. Hence, we carried out this study to know whether the benzofuran compound 3, 4-dihydro 4-oxo-benzofuro (3, 2-d) pyrimidine-2-propionic acid has got anti-inflammatory activity against chronic inflammation. Methods: Wistar albino rats were treated with benzofuran compound under study and phenylbutazone in the dose of 100 mg\kg, orally with 2% gum acacia as suspending agent and the effects were observed in chronic experimental model of inflammation namely, cotton pellet induced granuloma model. Results: In the present study, it was shown that the benzofuran compound under study has got significant anti-inflammatory activity against the chronic model of inflammation. Conclusion: Our experiment shows that the benzofuran compound under study has got significant anti-inflammatory activity and may, as well become an additional anti-inflammatory drug if further studies are conducted in this direction involving human beings.

Anti-Inflammatory Activities of Methanol Leaf Extract of Bridelia micrantha (Hochst) Baill. (Euphorbiaceae) in Wistar Rats.

Aerial parts of Bridelia micrantha, a semi-deciduous tree are widely used in African traditional medical practice in the treatment of painful inflammatory conditions of the joints. This study evaluated the anti-inflammatory activities of the methanol leaf extract of Bridelia micrantha using acute, sub-acute and chronic models of inflammation in Wistar rats. In the carrageenan-induced acute inflammation model, 400 mg/kg of extract produced 71.79 % edema inhibition while 200 mg/kg of the extract produced 55.13 % inhibition relative to 56.41 % inhibition of the rat paw edema with 200 mg/kg of Acetylsalicylic Acid (ASA) within 5 h. In the histamineinduced rat paw edema model, the extract exhibited 72.97 % protection at 400 mg/kg compared to 83.33 % edema inhibition with phenylbutazone (100 mg/kg) after 6 h. In the sub-acute model using formaldehyde-induced paw edema, 400 mg/kg of extract showed 59.77 % (0.35±0.03) inhibition after 24 h, while 54.02 % inhibition was produced by 200 mg/kg of extract and 200 mg/kg of ASA produced 56.32 % (0.38±0.04) inhibition when compared with the negative control group (0.87±0.05). In the cotton pellet-induced granuloma test, 400 mg/kg of extract gave 52.55 % (40.57±1.3) protection, while 200 mg/kg extract gave 47.25 % protection and 200 mg/kg ASA gave 49.38 % (43.25±1.8) when compared with normal saline treated group (85.5±3.2) after 7 days. The results obtained in this study showed that Bridelia micrantha leaf extract exhibited potent anti-inflammatory activities thus authenticating its acclaimed anti-inflammatory efficacy. It was concluded that the extract of Bridelia micrantha may be a potential anti-inflammatory agent in alleviating edema associated with arthritis and musculo-skeletal pains in humans.

Muerte súbita asociada a hipersensibilidad tipo I en un equino pura sangre de carrera / Sudden death associated to type I hypersensitivity in a racing horse

El objetivo de este estudio fue reportar un caso de hipersensibilidad tipo I con muerte súbita en un equino Pura Sangre de Carrera en el Hipódromo “La Rinconada” Caracas, Venezuela. Se tomaron muestras de sangre y orina para estudios toxicológicos mediante la técnica de ELISA competitivo. Se le práctico la técnica de necropsia, fueron colectadas muestras de musculo, tejido pulmonar, hepático, renal, gástrico, esplénico, corazón y sistema nervioso central para estudio histopatológico, las muestras fueron procesados por los métodos convencionales histológicos. Los hallazgos de necropsia fueron flebitis severa en vena yugular derecha, con hematoma en el surco yugular. Edema severo de glotis, edema, congestión y hemorragia pulmonar. Hemorragia petequial subendocardica. Bazo esplenocontraido y con focos de necrosis de coagulación. Hidronefrosis aguda con hematuria. Hígado con patrón lobulillar acentuado. El resto de los órganos con evidente congestión y hemorragia. Los cortes histológicos evidenciaron edema, congestión y hemorragia pulmonar severa. Hemorragia subepicardica marcada. Edema subcapsular esplénico y necrosis centro-folicular. Degeneración hidropica tubular, necrosis tubular aguda. Necrosis de corteza renal. Los estudios toxicológicos permitieron la detección de furosemida y fenilbutazona en las muestras de sangre y orina. En conclusión se reporta un síndrome de hipersensibilidad tipo I asociado a la administración de un producto comercial a base de Vitamina E 80mg, Pangamato sódico (B15) 1 mg, Selenio Sódico 0.6 mg, Antioxidantes y Vehículos Solubles c.s.p. con colapso, shock y muerte aguda en un equino Pura Sangre de Carrera mediante un estudio multidisciplinario clínico, anatomopatologico y toxicológico.

The aim of this study was to report a case of type I hypersensitivity to sudden death in a Thoroughbred race horses at the Hippodrome “La Rinconada” Caracas, Venezuela. Samples of blood and urine for toxicology studies using competitive ELISA. He practiced the technique of necropsy, samples were collected from muscle, lung tissue, liver, kidney, stomach, spleen, heart and central nervous system for histopathological examination, samples were processed by conventional histological methods. Autopsy findings were severe phlebitis right jugular vein, with hematoma in the jugular groove. Severe edema of glottis edema, pulmonary congestion and hemorrhage. Subendocardial petechial hemorrhage. Esplenocontraido Spleen foci of necrosis and coagulation. Hydronephrosis with acute hematuria. Liver accentuated lobular pattern. The rest of the organs with obvious congestion and hemorrhage. The histological sections showed edema, severe pulmonary congestion and hemorrhage. Marked subepicardial hemorrhage. Edema and necrosis subcapsular splenic follicular center. Tubular hydropic degeneration, acute tubular necrosis. Necrosis of renal cortex. Toxicological studies allowed the detection of furosemide and phenylbutazone in samples of blood and urine. In conclusion we report type I hypersensitivity syndrome associated with the administration of a commercial product based Vitamin E 80mg, sodium pangamate (B15) 1 mg, 0.6 mg; Sodium Selenium, Soluble Antioxidants and Vehicle qs with collapse, shock and acute death in a race Thoroughbred horses by a multidisciplinary clinical, pathological and toxicological.

Effects of nonsteroidal anti-inflammatorydrug on prevalence of helicobacter like organisms in gastric mucosa of thoroughbreds horses

The aim of this study was describe effects of NonsteroidalAnti-inflammatory drug on prevalence of Helicobacter LikeOrganisms in gastric mucosa of Thoroughbreds horses. Werestudied 54 Thoroughbred horses in the national race Track“La Rinconada” Caracas-Venezuela. All equine were treatedby seven days with phenylbutazone at an intravenous doseof 4.4 mg/kg. All horses presented Equine gastric ulcer syndromeacute superficial gastritis (25/54), chronic gastritis witherosion focal (16/54), chronic gastritis with erosion focal andulcers (14/54) in the gastric in both regions mucosa squamousregion and glandular regions (fundus). Helicobacter Like Organismsinfection in the stomach was confirmed by Warthin-Starry (38/54). Gastric mucosa revealed numerous spiralshapedbacteria morphologically resembling Helicobacter LikeOrganisms in squamous regions, margo plicatus (20/38) andnumerous spiral-shaped bacteria in fundic glands (18/54). Inconclusion, we detected high presence of Helicobacter LikeOrganisms in the gastric mucosa of Thoroughbred horse’streatment with phenylbutazone.

Intoxicación con fenilbutazona en un equino: reporte de un caso / Toxicity with phenylbutazone in a horse: a case report

Se describe el caso de un equino que desarrolló graves lesiones digestivas después de recibir dosis altasde fenilbutazona (FBZ) para tratar una claudicación. Al momento de la consulta tenía 9 días de evolución.Desde su llegada al hospital, se observó cojera grave de las cuatro extremidades, deshidratación y diarreafétida. Luego del examen físico, la anamnesis y las ayudas diagnósticas se propuso un dictamen de laminitis traumática, gastritis ulcerativa y colitis por intoxicación con antiinflamatorios no esteroides (AINES). Lacondición empeoró a pesar de la terapia y cuando se presentaron signos neurológicos se sugirió la eutanasia.Durante la necropsia se observaron lesiones graves en el tracto gastrointestinal, cascos y encéfalo. El objetivo de este artículo es describir la sintomatología, terapia y evolución de un paciente intoxicado con aines.

It is described a clinic case of an equine that developed severe digestive lesions after taking high dosage ofphenylbutazone to treat a lameness. At the moment of checking, it had nine days of evolution. Since its arrivingto the hospital, it was seen an intense lameness of the four limbs, dehydration and fetid diarrhea. After the physicexam, the interrogatory and the diagnostic aids it was proposed a diagnosis of traumatic laminitis, ulcerative gastritis and colitis by intoxication with Non-steroidal anti-inflamatory drug (NSAIDs). The condition became worse despite the therapy and when the neurological signs were presented. It was suggested the euthanasia. During the necropsy, it was seen severe lesions in the gastrointestinal tract, hooves and brain. The objective of this article is to describe la symptomatology, therapy and evolution of the intoxicated patient with NSAIDs.

Descreve-se o caso de um cavalo que desenvolveu lesões digestivas graves após receber altas doses defenilbutazona (FBZ) para tratar uma claudicação. No momento da consulta havia 9 dias de evolução. Desdesua chegada ao hospital, observou-se manqueira grave nas quatro extremidades, desidratação e diarréia fétida. Após o exame físico, a anamnese e os meios diagnósticos concluiu-se se tratar de laminite traumática, gastriteulcerativa e colite por intoxicaçãocom anti-inflamatórios não esteróides (AINES). A condição piorou apesardo tratamento e, quando apresentou sinais neurológicos, sugeriu-se a eutanásia. Na necrópsia observaram-selesões graves no trato gastrointestinal, cascos e enféfalo. O objetivo deste trabalho é descrever os sintomas, otratamento e a evolução de um paciente intoxicado com AINES.

Efeitos da fenilbutazona na cicatrização de feridas cutâneas experimentais em equinos / Phenylbutazone effects on experimental wound healing in horses

No processo de cicatrização por segunda intenção de feridas cutâneas experimentalmente induzidas em equinos, avaliaram-se os efeitos da fenilbutazona e comparou-se a cicatrização entre as regiões torácica e lombar. Utilizaram-se dez equinos, dos quais se retirou fragmentos circulares de pele de dois centímetros de diâmetro das regiões lombares e torácicas direita e esquerda. Os equinos foram distribuídos em dois grupos, sendo o primeiro controle, recebendo água destilada a cada 12 horas, durante cinco dias. O outro grupo foi tratado com fenilbutazona (4,4 mg/kg) com o mesmo intervalo e período do grupo controle. As feridas foram tratadas diariamente com Líquido de Dakin, momentos quando se procederam as observações macroscópicas. A cada 72 horas procederam-se as mensurações das feridas. Para análise histológica realizou-se biópsias no sexto e décimo quinto dia. O tempo total de reparo das feridas no grupo tratado foi maior em aproximadamente 12 dias (37 dias para o grupo controle e 49 dias para o grupo tratado). Não se observou diferença significativa do tempo de cicatrização entre as feridas torácicas e lombares de um mesmo grupo. As avaliações macroscópicas e histopatológicas mostraram o efeito inibidor da fenilbutazona quando comparada com o grupo controle na cicatrização de feridas cutâneas por segunda intenção em equinos.

The purpose of this study was to investigate phenylbutazone effects on second intention wound healing, and to compare the healing process between the thoracic and lumbar areas. Ten horses were submitted to circular full-thickness wound produced on both sides of the thoracic and lumbar areas. Animals were gathered into two experimental groups, one receiving daily IV injections of phenylbutazone (4,4mg/kg) and the other (control group) distillated water for five days. All wounds were daily treated with local Dakin's solution. The wound contraction rates were determined by serial measurements each 72 hours. At the 6th and 15th post surgical days, biopsies were performed for histological analysis. Thoracic and lumbar wound contraction was decreased in the phenylbutazone group. The time to complete healing was significantly greater in phenylbutazone group (49 days) than in control group (37 days). There was no significant difference between thoracic and lumbar area in the same group. Gross and histopathology analysis showed the inhibitory effect of phenylbutazone on the second-intention wound healing when compared to the control group.

Molecular pharmacological interaction of phenylbutazone to human neutrophil elastase

Human neutrophil elastase (HNElastase, EC 3.4.21.37), a causative factor of inflammatory diseases, was purified by Ultrogel AcA54 gel filtration and CM-Sephadex ion exchange chromatography. HNElastase was inhibited by phenylbutazone in a concentration dependent manner up to 0.4 mm, but as the concentration increased, the inhibitory effect gradually diminished. Binding of phenylbutazone to the human neutrophil elastase caused strong Raman shifts at 200, 440, and 1194 cm-1. The peak at 1194 cm-1 might be evidence of the presence of -N=N-PHI radical. The core area of the elastase, according to the visual molecular model of human neutrophil elastase, was structurally stable. A deeply situated active center was at the core area surrounded by hydrophobic amino acids. Directly neighboring the active site was one positively charged atom and two atoms carrying a negative charge, which enabled the enzyme and the drug to form a strong interaction. Phenylbutazone may form a binding, similar to a key & lock system to the atoms carrying opposite charges near the active site of the enzyme molecule. Furthermore, the hydrophobicity of the surrounding amino acid near the active site seemed to enhance the binding strength of phenylbutazone. Binding of phenylbutazone near the active site may cause masking of the active site, preventing the substrate from approaching the active site and inhibiting elastase activity.