Profiling of commmercial agave fructooligosaccharides using ultrafiltration and high performance thin layer cromatography
Perfil de fructooligosacáridos de agave comerciales empleando ultrafiltración y cromatografía en capa fina de alta resolución
C. Alvarado, R.M. Camacho*, R. Cejas, J.A. Rodríguez
Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco Av. Normalistas 800 Col. Colinas de la Normal, CP.44270, Guadalajara, Jalisco México. *Corresponding author. E-mail: rcamacho@ciatej.mx Tel. 01-33-33455200 Ext. 2130, Fax 01-33-33455245
]]> Received August 6, 2013.
Abstract
A set of ingredients from chicory and Agave were analyzed to obtain the carbohydrate distribution and band profiling through Ultrafiltration and High Performance Thin Layer Chromatography (HPTLC). For method standardization, reference materials and ingredients were used. Carbohydrate distribution was based in their separation by Ultrafiltration through 3 kDa membrane, allowing separation and quantification of long-chain fructans (degree of polymerization DP>10) from short-chain fructans (FOS DP<10). The relation of long-chain fructans versus short-chain fructans (long:short) resulted in 52-60:40-48 for Agave materials showing almost the same relation as the chicory ingredients. The retention factor (Rf) of each band by HPTLC from each sample was recorded and compared to generate a characteristic chicory and Agave profile. The bands of fructose, sucrose, 1-kestose and nystose were identified in all materials analyzed. Additionally, at least four distinctive bands were observed in chicory and three in the Agave materials. It was possible to differentiate between sources of fructans, since the band profiles were different. This approach to ingredients study showed to be useful to discriminate between Agave and chicory fructans and for following food industry quality control.
Keywords: fructans, fructooligosaccharides, Agave FOS, thin layer chromatography, nystose, kestose.
Resumen
Un grupo de ingredientes de achicoria y Agave se analizaron para obtener la distribución de carbohidratos y el perfil de bandeo mediante Ultrafiltración y Cromatografía en Capa Fina de Alta Resolución (HPTLC). La estandarización del método se realizó empleando materiales e ingredientes de referencia. La distribución de carbohidratos se basó en su separación por Ultrafiltración con membranas de 3 kDa, permitiendo la separación y cuantificación de fructanos de cadena larga (grado de polimerización (DP)>10) de los de cadena corta (FOS DP<10). La relación fructanos largos respecto de los cortos (largo:corto) resultó de 5260:40-48 para materiales de Agave, mostrando una relación muy parecida a los de achicoria. Los factores de retención (Rf) de cada banda por HPTLC para cada muestra fueron registrados y comparados para generar un perfil característico para achicoria y Agave. Las bandas de fructosa, sacarosa, 1-kestosa y nistosa se identificaron en todos los materiales. Además se encontraron al menos cuatro bandas exclusivas en achicoria y tres en Agave. Fue posible diferenciar entre los orígenes de los fructanos ya que el perfil de bandeo fue diferente entre ingredientes. Este enfoque mostró ser útil para el estudio de ingredientes en la discriminación entre fructanos de Agave y achicoria así como para el control de calidad en la industria de alimentos.
Palabras clave: fructanos, fructooligosacáridos, FOS de Agave, cromatografía en capa fina, nistosa, kestosa.
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Acknowledgment
Authors thank for financial support through SAGARPA grant 109799.
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