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Hamilton produces the Polymeric based PRP-X400 HPLC columns for the analysis of Glyphosateand its metabolite in drinking water. It can also be used for Inositol and sugaralcohols and some uniquehydrophilic interaction separations.

PRP-X400 Advantages for Glycophosphate Analysis

Faster Separation
The PRP-X400 column provides a faster separation than the Aminex Glyphosate Column.

Use at Room Temperature
The PRP-X400 column does not have to be heated to 65ºC, so you don't need to purchase a column heater for this method.

No Methanol Required
PRP-X400 columns do not require the use of methanol in the mobile phase.

Lower Cost
PRP-X400 columns cost much less than other glyphosate columns.

LITERATURE REFERENCES PRP-X400

Glyphosate and it's Metabolite at 100 ppb Glyphosate100 ppb AminomethylphosphonicAcid 100 ppb Conditions:5.0 mMMonobasic Potassium Phosphate pH 2.1. Isocratic. Ambient. Flow:0.5mL/min. Injection:100 µL Detection:Fluorescenceex: 340 nm, em: 455 nm Post Column Conditions: Oxidation with 15 ppm Calcium hypochlorite at 0.2 mL/min.; Derivatization with OPA at 0.3 mL/min.

Use the PRP-X400 Column for EPA Method #547

SeparationMechanism
ThePRP-X400 is a 7 µm poly(styrene-divinylbenzene) sulfonate cation exchangesupport (2.5 meq/gm) column. It separates glyphosate and aminomethylphosphonicacid according to charge in less than 10 minutes. This separation requirespost-column oxidation and derivatization (See Figure 1).

MobilePhase Preparation
Toprepare 0.005 M monobasic potassium phosphate (KH2PO4) pH 1.9, dissolve 0.68 gmof monobasic potassium phosphate in 1 Liter of deionized water. Adjust the pHto 1.9 using concentrated phosphoric acid. Prior to using this preparation,filter it through a 0.45 µm nylon filter and degas.

Detection
Postcolumn reaction (oxidation) with calcium hypochlorite followed byderivatization with o-phthalaldehyde solution provides sensitive (6 ppb orlower) and selective (primary and secondary) amine detection. To achievelow-level detection (6 ppb) of glyphosate and its metabolite, follow theinstructions listed under Separation Conditions.

Separation Conditions

Separation Procedure

OxidationSolution Preparation (15 ppm calcium hypochlorite)
StockConcentrate Solution Preparation - To prepare the 1500 ppm concentratesolution, add 0.23 gm of tech grade calcium hypochlorite to 100 mL of deionizedwater. With a 2 µm nylon filter, remove any insoluble calcium carbonate (as itproduces a cloudy solution). Store the solution in the freezer. The shelf lifeis several freeze/thaw cycles.

WorkingOxidation Solution Preparation
Dissolve1.36 gm monobasic potassium phosphate, 11.60 gm sodium chloride, and 0.40 gmsodium hydroxide (or use 0.50 mL 50% w/w sodium hydroxide solution) in 950 mLdeionized water. Add 10 mL of 1500 ppm calcium hypochlorite stock concentratesolution, and dilute to 1 Liter. Filter through a 0.45 µm nylon filter. Preparethis solution fresh daily. Use and store this solution in an inert atmosphere(helium or nitrogen). Degas before use.

Figure 1

Derivatization Solution Preparation (o-phthalaldehyde)
Dissolve 19.1 gm of disodium tetraborate decahydrate in 950 mL of deionized water. Heat the solution to approximately 50ºC for about one hour to dissolve the disodium tetraborate decahydrate (or prepare the solution one day in advance and allow the borate to dissolve). Cool the solution to room temperature, and adjust the pH to 10.9 with 1 N sodium hydroxide. Now dissolve 0.80 gm phthalic dicarboxyaldehyde (Aldrich Part # P3,940-0) in 10 mL methanol. Add all 10 mL to the disodium tetraborate decahydrate solution. Then add 50 µL of 2-mercaptoethanol. (Caution: Use adequate ventilation and /or a hood when handling 2-mercaptoethanol, as the fumes are noxious.) Dilute the concentrate to 1 Liter with deionized water, mixing well. Filter the mixed solution through a 0.45 µm nylon filter, and degas before using. Store the solution in an inert atmosphere (helium or nitrogen). Refrigerate the unused solution. Shelf life is one or two weeks.

Reaction coils can be made in the laboratory (see Separation Conditions) or entire post-column derivatization systems are available for purchase.

Any further questions? Please contact our Customer Care Centre:onlinestore@carlstuart.com

PRP-X400COLUMNS

4001LiquidChromatography and postcolumn indirect detection of Glyphosate.
Lovdahl,M.J.; Pietrzyk, D.J.; J. Chromatography,1992, 602, 197.

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