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Glyphosate is an herbicide that has been on the market since the 1970s and has since become the most commonly used weedkiller. The use of glyphosate is highly controversial among experts, users and consumers, as it is suspected of being carcinogenic.
Various liquid chromatography separation techniques can be used to analyse glyphosate and its metabolites (e.g. aminomethylphosphoric acid - AMPA). These include ion chromatography, hydrophilic interaction chromatography (HILIC) and reversed-phase chromatography. Due to the high polarity of glyphosate and its UV inactivity, the amino nitrogen is often derivatised with an Fmoc protecting group during sample preparation, which facilitates the separation and detection of the resulting substances.
1. AMPA 2. glufosinates 3. glyphosates 4. MPPA (3-methylphosphinicopropionic acid)
Column: HILICpak VT-50 2D 150x2.0mm, 5µm
Mobile phase: 70/20/10 water/1% formic acid/acetonitrile
Flow rate: 0.3 mL/min
Detection: ESI-MS (SIM)
Temperature: 40 °C
Sample: 1 µg/mL
Injection volume: 5 µL
1. glufosinates 2. glyphosates
Column: IC SI-90 4E 250x4.0mm, 9µm
Mobile phase A: 0.3 mM NaOH
Mobile phase B: 0.3 mM NaOH + 20 mM sodium carbonate
Gradient:
Time / min | %A | %B |
0 | 95 | 5 |
20 | 95 | 5 |
30 | 50 | 50 |
40 | 50 | 50 |
Flow rate: 1 mL/min
Suppressor: Dionex AMMS-ICS II
Detector: ICP/MS, P at m/Z=31
1. glyphosates
Column: Inertsil ODS-4 150x4.6mm, 5µm (5020-03945)
Mobile phase A: Acetonitrile
Mobile phase B: 50 mM potassium dihydrogen phosphate (pH 2.5 with phosphoric acid)
Gradient: 35%A - 2.5 min, 70%A - 3 min, 35%A - 15.5 min
Flow rate: 1 mL/min
Temperature: 40 °C
Detection: Fluorescence excitation 270 nm, emission 315 nm
Injection volume: 10 µL
HPLC - Reversed Phase:
HPLC - IEX:
HPLC - Reversed Phase:
HPLC - Mixed-Mode:
HPLC - Reversed Phase:
HPLC - HILIC:
HPLC - HILIC:
HPLC - Ion chromatography:
HPLC - Ion chromatography:
HPLC - Reversed Phase
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