Team:Bielefeld-Germany/HPLC isocratic

From 2011.igem.org

(Difference between revisions)
(→HPLC method)
(→HPLC method)
 
(6 intermediate revisions not shown)
Line 1: Line 1:
===HPLC method===
===HPLC method===
* C18 reverse phase column
* C18 reverse phase column
-
* Isocratic method: 45 % [[Team:Bielefeld-Germany/Protocols#Used_chemicals | acetonitrile]]
+
* Isocratic method: 45 % [[Team:Bielefeld-Germany/Protocols#Used_chemicals | Acetonitrile]]
-
* Flow = 0.6 mL/min
+
* Flow = 0.6 mL min<sup>-1</sup>
* UV-detection at 227 nm
* UV-detection at 227 nm
 +
* Internal standard: 100 mg L<sup>-1</sup> [[Team:Bielefeld-Germany/Protocols#Used_chemicals | Bisphenol F]] (BPF)
* Column:
* Column:
-
** Eurospher II 100-5 C18p
+
** Eurospher II 100-5 C18p by [http://www.knauer.net/ Knauer]
** Dimensions: 150 x 4.6 mm with precolumn
** Dimensions: 150 x 4.6 mm with precolumn
** Particle size: 5 µm
** Particle size: 5 µm
** Pore size: 100 Å
** Pore size: 100 Å
** Material: silica gel
** Material: silica gel
 +
* Software:
 +
** Clarity (Version 3.0.5.505) by [http://www.dataapex.com/ Data Apex]
 +
* Autosampler:
 +
** Midas by [http://www.spark.nl/ Spark Holland]
 +
** Tray cooling: 10 °C
 +
* Pump:
 +
** L-6200A Intelligent Pump by [http://www.hitachi.com/ Hitachi]
 +
* UV-Detector:
 +
** Series 1050 by [http://www.hp.com/ Hewlett Packard]
 +
 +
[[File:Bielefeld2011 BPA + Abbauprodukt-2.JPG|900px|left|thumb|Chromatogram obtained using the above described materials and methods. Three distinctive peaks are of interest: BPA at 7,3 min retention time, BPF at 5,8 min retention time and the degradation product of BPA at 4,6 min retention time.]]

Latest revision as of 13:54, 31 August 2011

HPLC method

  • C18 reverse phase column
  • Isocratic method: 45 % Acetonitrile
  • Flow = 0.6 mL min-1
  • UV-detection at 227 nm
  • Internal standard: 100 mg L-1 Bisphenol F (BPF)
  • Column:
    • Eurospher II 100-5 C18p by [http://www.knauer.net/ Knauer]
    • Dimensions: 150 x 4.6 mm with precolumn
    • Particle size: 5 µm
    • Pore size: 100 Å
    • Material: silica gel
  • Software:
    • Clarity (Version 3.0.5.505) by [http://www.dataapex.com/ Data Apex]
  • Autosampler:
    • Midas by [http://www.spark.nl/ Spark Holland]
    • Tray cooling: 10 °C
  • Pump:
    • L-6200A Intelligent Pump by [http://www.hitachi.com/ Hitachi]
  • UV-Detector:
    • Series 1050 by [http://www.hp.com/ Hewlett Packard]
Chromatogram obtained using the above described materials and methods. Three distinctive peaks are of interest: BPA at 7,3 min retention time, BPF at 5,8 min retention time and the degradation product of BPA at 4,6 min retention time.
Retrieved from "http://2011.igem.org/Team:Bielefeld-Germany/HPLC_isocratic"