The Application Notebook
Allantoin is a heterocyclic organic compound derived from purine.
Michael Halvorson1, Torsten Kretschmer2, and Martin Roedel2, 1Gilson, Inc., and 2Macherey-Nagel GmbH
Allantoin is a heterocyclic organic compound derived from purine. Allantoin has a long history of use in a variety of topical pharmaceuticals and cosmetics for skin care due to its keratolytic, moisturizing, soothing, and anti-irritant properties. Allantoin is typically used in these products at a level of 0.1 to 2.0%. Solid phase extraction (SPE) is often used as a purification step prior to analysis of allantoin in these products due to the complex nature of the matrices.
This study describes an automated SPE protocol using a Gilson GX-271 ASPEC™ System for the extraction of allantoin from cosmetics prior to analysis by HPLC using a NUCLEODUR® 100-3 HILIC column.
Mix 1 g of sample containing allantoin with 100 mL of ultra-pure water. Allantoin-free cosmetics/topicals were spiked with 5 mg allantoin.
The SPE procedure used 3 mL Macherey-Nagel CHROMA-BOND®HR-XA (60 mg) cartridges.The SPE protocol is entirely automated using the Gilson GX-271 ASPEC system. The SPE steps are summarized with the schematic provided in the GX-271 ASPEC control software, Trilution LH™ (Figure 1).
Figure 1: Trilution LH SPE and liquid handling tasks for extraction of Allantoin.
Column: Macherey-Nagel EC 125/3 NUCLEODUR® 100-3 HILIC (Part no. 760 531.30)
Eluent A: 10 mmol/L Ammonium chloride, pH 3.0 20
Eluent B: Acetonitrile 80
Flow Rate: 0.3 mL/min
Injection Volume: 20 µL
Concentration: β(Allantoin) = 5 µg/mL eluent
Detection: UV, 214 nm
Figure 2: Chromatogram of Allantoin from cosmetic product following SPE extraction. The retention time for Allantoin is 3.66 min.
The recovery of allantoin from the cosmetic product was 85.5% (n = 3).
CHROMABOND and NUCLEODUR are registered trademarks of Macherey-Nagel GmbH. ASPEC and Trilution are trademarks of Gilson, Inc.
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