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Method Abstract
for Post-column Liquid Chromatography 104
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Polyether antibiotics, such as
Monensin, Narasin, and Salinomycin, in raw
material, premix, liquid supplements, and final
feeds, are best quantified using a
high-performance liquid chromatograph equipped
with a post-column reactor. The polyether
antibiotics react, post-column, with Vanillin in
the presence of Sulfuric acid, and the resulting
products can be detected at 520 nm. However, this
corrosive reagent can adversely degrade most
instruments. Pickering’s special Post- Column
Systems, comprised of non-metallic parts and
washing pump head, offers resistance to sulfuric
acid. Special operating instructions will also be
included with this instrument. Contact Pickering
Laboratories for more information about the Inert
Vector PCX Post-Column System for polyether
antibiotics.
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COLUMN: |
Polyether reversed-phase
column, C18, 4.6 x 250 mm, Catalog
No. 2381750 |
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TEMPERATURE: |
40 °C |
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FLOW RATE: |
0.7 mL/min |
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MOBILE PHASE: |
Methanol/5% acetic acid
in water (90/10),
Isocratic |
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POST-COLUMN SYSTEM: |
Vector PCX or Pinnacle
PCX (Non-metallic post-column
derivatization system is required) |
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REAGENT 1: |
Concentrated Sulfuric
acid/Methanol (4:96 v/v) |
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REACTOR 1: |
ambient, 0.1 mL |
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REAGENT 2: |
60 g of Vanillin (or p-dimethylaminobenzaldehyde)
in 950 mL of Methanol |
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REACTOR 2: |
90 °C, 1.4 mL |
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FLOW RATE: |
0.3 mL/min |
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DETECTION: |
UV-Vis detector:
Vanillin  =520
nm, DMAB =450
nm |
REFERENCES
1. M.R.
LaPointe and H. Cohen, J.A.O.A.C., 71 (1988)
480–484.
2. J.M.
Rodewald, J.N. Moran, A.L. Donoho, and M.R.
Coleman, J.A.O.A.C.,75 (1992) 272–279.
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