title: Validated reversed phase-high performance liquid chromatography-diode array detector method for the quantitation of Rutin, a natural immunostimulant for improving survival in aquaculture practice, in toonea sinensis folium.
abstract: BACKGROUND
  Rutin is a bioflavonoid of strong immunostimulating activity from the Toonea Sinensis Folium, which has shown a significant ability to increase the survival rate of white shrimp with bacterial infection. However, no method for the quantitation of this active ingredient in the herb has been reported to date.
  
  
  METHODS
  A reversed phase-high performance liquid chromatography-diode array detector (RP-HPLC-DAD) method was developed to quantify Rutin in the Toonea Sinensis Folium, with the HPLC conditions optimized, followed by validation for linearity, accuracy, precision, limit of detection (LOD), repeatability, and stability. Then, the established method was used to determine the content of Rutin in two samples.
  
  
  RESULTS
  The separation was performed on a Waters XBridge Shield RP18 column (150 mm × 4.6 mm, 5 μm) kept at 25°C, and acetonitrile and water containing 0.1% acetate acid (18:82, v / v)-composed mobile phase was constantly driven at 1.0 mL / minute during the analysis. Twenty microliters of sample solution or standard solution were injected into the HPLC system and 254 nm was selected to monitor the separation. A strong linear relationship between the peak area and concentration of Rutin was observed within the range of 0.01044 - 0.2610 mg / mL (r(2) = 1.0000). The LOD was 0.03915 μg / mL, and recovery of Rutin was from 97.6 to 99.6%. In addition, the method was also validated to be repeatable, stable, precise, and accurate.
  
  
  CONCLUSIONS
  An efficient and reliable RP-HPLC-DAD method was established, which could be used for routine analysis of Rutin in Toonea Sinensis Folium and to assist in the quality control of this herb.
authors.affiliation: Department of Chinese Medicine and Pharmacy, School of Pharmacy, Jiangsu University, China.
authors.forename: Yuping
authors.initials: Y
authors.lastname: Shen
authors.forename: Huawu
authors.initials: H
authors.lastname: Yin
authors.forename: Bin
authors.initials: B
authors.lastname: Chen
authors.forename: Guohua
authors.initials: G
authors.lastname: Xia
authors.forename: Huan
authors.initials: H
authors.lastname: Yang
authors.forename: Xiaobin
authors.initials: X
authors.lastname: Jia