Yanming  Wei, Guli  Asa, Chun  Zhang, Yonggang  Wang, Meng  He, Yilong  Yin, Xiping  Fan, 
    Study for High Throughput Anthrone-Sulfuric Acid Method for Determination of Polysaccharides,
    Journal of Animal and Veterinary Advances,
    Volume 12,Issue 19,
    2013,
    Pages 1504-1511,
    ISSN 1680-5593,
    javaa.2013.1504.1511,
    (https://makhillpublications.co/view-article.php?doi=javaa.2013.1504.1511)
    Abstract: A rapid, sensitive and high throughput micro-plate method was established 
  to quantify polysaccharides in this study. Anthrone-sulfuric acid micro-plate 
  assay in a certain reaction was tested. A single test and a response surface 
  methodology was applied to design and optimize four factors including reaction 
  time, incubation temperature, anthrone concentration and the amount of anthrone 
  in order to maximize R<SUP>2</SUP> and minimize RSD in each standard curve which 
  obtained in different conditions. It was demonstrated that the order of significant 
  impacting R<SUP>2</SUP> is incubation temperature, anthrone concentration, reaction 
  time and the quantity of anthrone add to while the order of significant impacting 
  RSD is incubation temperature, anthrone concentration, the amount of anthrone 
  and reaction time. Preferred reactive condition is be discovered and the best 
  incubation temperature is 96&deg;C, the best reaction time is 36 min, anthrone 
  concentration is 1.1 mg mL<SUP>-1</SUP> and the amount of anthrone is 150 &#956;L. 
  The detectable range of this method is 20-260 &#956;g L<SUP>-1</SUP> under the 
  optimized condition. Recovery test showed high accuracy and practicability in 
  this method by recovery rate ranged in 99.5-100.6%. The method was successfully 
  applied to quantify polysaccharides of different source and the method has high 
  resistance to interference of protein during measurement process.
    Keywords: Micro-plate;anthrone-sulfuric acid;polysaccharide;response surface methodology;recovery test