Gas chromatographic method for determining very long chain fatty acids that compose D003 in 1 to 10 mg/ml suspensions used in pharmacological and toxicological studies

D003 is a mixture of fatty acids (C240 to C36.0), that shows antiplatelet, antithrombotic and cholesterol-lowering effects in experimental models. A gas chromatographic method using a DB-5 Widebore column and 1-nonadecanoic acid as internal standard was developed and validated in order to determine D003 in the aqueous suspensions, with concentrations from I to 10 mg/mL, used in pharmacological and toxicological studies . Fatty acids were analyzed as methyl esters derivatives, prepared using 5  aqueous HCI-methanol . This  method was specific for D003 determination in these suspensions . The determination of total content of fatty acids showed a CV of 0.39%. The values of the correlation coefficients (r) were higher than that of the acceptation limit (0 .99) for the individual fatty acids and for the total content of them, indicating that correlations weree positives for a probability higher than 99.9%. In the repeatability study the CV for total content of D003 (1.55, 1.68 and 0.62%, respectively) were lower, indicating that the method has repeatability. The inter-mean precision for the total content of fatty acids (CV 1 .47%), demonstrated that the method is reproducible . The average recovery obtained for the total content of acids was greater than 99% for all cases. Also, for all the fatty acids 4,o < tab for a 95% of releability, there were no significant differences between the average recovery and 100%, then, it can be considered that this method is exact. The method was suitable for quality control and stability studies of these suspensions

Determination_of D003 by capillary gas chromatography

D003 is a mixture of saturated very long-chain fatty acids ranging from 24 to 36 carbon atoms, obtained from sugar cane (Saccharum officinarum L.) wax, with antiplatelet, 'antithrombotic, antioxidant and cholesterol-lowering effects . A capillary gas chromatographic method for the determination of this product was developed and validated . The acids were analyzed as methyl ester derivatives and separated in a BP-5 widebore column using 1-nonacosanoic acid as internal standard . A good linearity (r > 0 .99; CVs of the response factors and of the slopes lower than 5 and 2 %, respectively) ; as well as absence of bias was found for both, total and individual acids in the whole studied concentration range, from 30 to 150 % of the nominal mass . Quantitative recoveries (98 .9 and 100 .62 %) and good intermediate precision (CV = 0 .96 %) were also obtained . The specificity study showed no interferences regarding the determination of this mixture, once the samples were submitted to stress conditions. According to these results, it can be ensure that the validated procedure is appropriate for the quality control and stability studies of this product

Estudio de estabilidad de tabletas que contienen 20 mg de policosanol como ingrediente activo

El objetivo de un estudio de estabilidad es determinar el período de tiempo durante el cual el producto farmacéutico conserva adecuadamente sus propiedades al ser almacenado en determinadas condiciones. El estudio de estabilidad de las tabletas que contienen 20 mg de policosanol, un nuevo medicamento con propiedades hipocolesterolemizantes, se realizó con el objetivo de predecir la fecha de vencimiento e investigar la posible aparición de productos de degradaci ón. Para ello, se observaron durante el estudio propiedades como: color, dureza, desintegración, masa promedio, contenido de policosanol y contenido de microorganismos. El contenido de ingrediente activo se determinó por cromatografía gaseosa, utilizando la técnica validada a tales efectos. También fueron estudiados los efectos de las condiciones drásticas de almacenamiento tales como: hidrólisis ácida y básica, oxidación, fotólisis y termólisis. Por otra parte, se realizaron estudios a largo plazo teniendo en cuenta dos zonas climáticas, o sea, zona II (25 °C y 60 % de humedad relativa (HR)) y la zona IV (30 °C y 70 % de HR) y un estudio acelerado a 40 °C y 75 % de HR. Estos estudios demostraron que esta formulación es estable, ya que no se produjeron cambios significativos en los indicadores estudiados según los criterios de aceptación establecidos en las condiciones estudiadas. El contenido de policosanol se mantuvo con muy poca variación durante todo el estudio, por lo que no fue posible utilizar el método cinético de Arrhenius para la determinación de la fecha de vencimiento

Stability studies of film-coated tablets containing 5 mg of D-003 as active ingredient

D-003 is a new under research drug with potential use for preventing or treating atherothrombotic diseases. This product is made up of a mixture of very high molecular weight aliphatic acids (C24:0 to C36:0) purified from sugar cane (Saccharum officinarum, L.) wax. The aim of this study was to determine the stability of the proposed formulation of D-003 5 mg film-coated tablets, following International Harmonization Conference guidelines for knowing their possible specifications change on storage conditions and for predicting their expiration date. Specifications such as D-003 content, color, tablet weight, hardness, disintegration time, and microbial content were assessed. Stress testing included acid hydrolysis, thermolysis, oxidation and photolysis. In addition, accelerated studies under drastic conditions: 40 oC and 75 % of relative humidity (RH) as well as long-term studies (48 months) at conditions of Climatic Zones IV (30 oC and 70 % RH) and II (25 oC and 60 % RH) were also conducted using three batches. Tablets were crushed to a fine powder and the active ingredient with chloroform was extracted in a hot way. The acids were converted to methyl esters and in a BP-5 wide-bore capillary column by gas chromatography were analyzed. Quantification by the internal standard method, using 1-nonacosanoic acid was carried out and peaks purity by gas chromatography - mass spectrometry was confirmed. No significant changes or trends either on the content of D-003 or on other specifications were found in any study

Five-year stability studies by GC and GC-MS of D-003 (a mixture of C24:0 to C36:0 fatty acids) alone and in 5 mg film-coated tablets

El D-003 es nuevo producto con efectos reductores del colesterol y antioxidantes. Se trata de una mezcla de ácidos grasos alifáticos de alto peso molecular (C a C ) purificados de la cera de caña de azúcar (Saccharum officinarum, L.), donde el ácido octacosanoico es el componente mayoritario. Se determinó la estabilidad del D-003 en estado natural y en forma de tabletas recubiertas de 5 mg, siguiendo las guías de la ICH. Los estudios de validación de los métodos por CG demostraron una buena linealidad (r > 0,9994; CVs de los factores de respuesta y de las pendientes 0.9994, RSDs of the response factors and of the slopes < 5 and 2%, respectively), high accuracy (recoveries from 98.9 to 100.6%) and precision (RSD < 2% for repeatability and reproducibility). Specifications such as content, color, tablet weight, hardness, disintegration time, and microbiological content were assessed. Stress testing included acid hydrolysis, thermolysis, oxidation and photolysis. In addition, accelerated studies (12 months) under drastic conditions: 40 °C and 75% of relative humidity (RH) as well as long-term studies (60 months) at conditions of Climatic Zones IV (30 °C and 70% RH) and II (25 °C and 60% RH) were also conducted using three batches of AI and tablets. No significant changes or trends neither on the content nor on other specifications were found in any study. Overall, D-003 is very stable as such and in 5 mg film-coated tablets, with a shelf life of 5 years in climatic conditions of Zones IV and II.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Physico-mechanical characterization and interactions with excipients of D-003, a mixture of fatty acids isolated from sugar cane wax

El D-003 es un nuevo producto que consiste en una mezcla de ácidos grasos de elevado peso molecular (desde C24:0 hasta C36:0), donde cada uno de ellos está en una proporción reproducible. Este producto presenta efectos antiplaquetarios y como reductor del colesterol. Como parte de los estudios de formulación del D-003 fueron llevadas a cabo la caracterización físico-mecánica y su interacción con excipientes para tabletas. El presente estudio utilizó la calorimetría diferencial de barrido (CDB) y la termogravimetría (TG) con el objetivo de evaluar la estabilidad térmica y las transiciones características del D-003 en presencia de diversos excipientes. Fueron medidas otras propiedades, tales como el ángulo de contacto, solubilidad, humectabilidad, distribución del tamaño de partículas, propiedades de fluido, densidades por asentamiento, por vertido y de alta presión, así como la reducción de volumen bajo compresión. Estos estudios, según nuestro conocimiento, no han sido previamente reportados para estos ácidos. Los estudios por CDB demostraron que no hubo interacción química o física entre el D-003 y los excipientes investigados. También, los análisis termogravimétricos probaron la elevada estabilidad térmica del D-003, el cual es estable y funde sin descomposición hasta temperaturas tan elevadas como 220 °C. Los lotes industriales estudiados mostraron adecuada pureza y distribución uniforme del tamaño de partícula, buenas propiedades de fluido, compresibilidad y cohesión, lo cual sustenta que sea un polvo apropiado para la elaboración de tabletas revestidas como forma terminada. Sin embargo, la muy baja solubilidad del D-003 en la mayoría de los disolventes, especialmente en agua, es el aspecto más crítico para el desarrollo la forma terminada, lo cual es importante para decidir la formulación del revestimiento.D-003 is a new product consisting of a mixture of fatty acids of very high molecular weight (from C24:0 to C36:0) in a reproducible proportion each, with cholesterol-lowering, and antiplatelet effects. As part of the formulation studies of D-003 a physico-mechanical characterization and its interaction with tablet excipients were carried out. The present study used differential scanning calorimetry (DSC) and thermogravimetry (TG) in order to evaluate the characteristic transitions and thermal stability of D- 003 in presence of several excipients. Other properties, such as the angle of contact, solubility, wettability, particle size distribution, flow properties, tapped, bulk and at high pressure densities, as well as volume reduction under compression were measured. These studies, to our knowledge, have not been previously reported to these fatty acids. DSC studies demonstrated that there was no chemical or physical interaction between D-003 and the investigated excipients. Also, thermogravimetric analysis proved the high thermal stability of D-003, which melts without decomposition and is stable at temperatures as high as 220 °C. Industrial batches of D-003 showed adequate purity and uniform particle size distribution, good flow properties, compressibility and compactability, which support a suitable powder for manufacturing film-coated tablets as finished forms. Nevertheless, the very low solubility of D-003 in most solvents, especially in water, is the most critical aspect for developing the finished form, being relevant to decide the coating formulation.Colegio de Farmacéuticos de la Provincia de Buenos Aire