Ion-pair reagents (IPR) are used in reversed-phase liquid chromatography (RP-LC) and has effectively separated ionic or ionizable analytes.
Successful applications of Ion-pair Reagents
Single- and double- stranded RNA / biological or synthetic (2011) [1]
Stationary Phase: Non-porous polymeric columns and monolithic polymeric capillary columns
Ion-Pair Reagent: Various traditional or volatile amine cation salts
Detector: Fluorescent-based assays or MS
Features:
-
High-resolution separations of short and long RNA molecules
-
Less than 10-minute separations
Metformin-alogliptin, and Metformin-repaglinide binary tablets (2019) [2]
Stationary Phase: Inertsil ODS column (250 mm, 4.6 mm, 5 μm)
Mobile Phase: sulfonateAcetonitrile; Phosphate buffer;Water
Ion-Pair Reagent: 0.3 % sodium heptane
Detector: UV
Features:
- Good recoveries achieved
- Sharp and well separated peaks
- Accurate, selective, and ecofriendly
- Method applicable in QC laboratories for the routine analysis of these drugs
Lactose, glucose, and galactose / original skim milk (2012) [3]
Stationary Phase: C18 column
Ion-Pair Reagent: Tetrabutylammonium hydrogen sulphate（CAS:32503-27-8）
Detector: UV and fluorescence
Features:
- Accurate and reproducible results
- Short retention times
- Simpler and quicker preparation compared to gas chromatography (no need to convert sugars into volatile derivatives)
- Low limit of quantification (LOQ) and limit of detection (LOD)
Nitrate and thiocyanate / natural water samples (2012) [4]
Stationary Phase: C18 column
Mobile Phase: 30:70 Methanol:Water (isocratic)
Ion-Pair Reagent: 0.08 mM Hexadecyltrimethyl-ammonium bromide (HTAB)
Detector: UV
Features:
- Simple
- Sensitive, accurate, and reproducible
Reference:
Dickman, M. J. Ion Pair Reverse-Phase Chromatography: A Versatile Platform for the Analysis of RNA. Chromatography Today. 2011. 22 – 26.Mahrouse, M. A., Lamie, N. T. Experimental design methodology for optimization and robustness determination in ion pair RP-HPLC method development: Application for the simultaneous determination of metformin hydrochloride, alogliptin benzoate and repaglinide in tablets. Microchemical Journal 2019, 147, 691-706.Erich, S., Anzmann, T., Fischer, L. Quantification of lactose using ion-pair RP-HPLC during enzymatic lactose hydrolysis of skim milk. Food Chemistry 2012,135(4), 2393-2396.Soleimani, M., Yamini, Y., Rad, F. M. A simple and high resolution ion-pair HPLC method for separation and simultaneous determination of nitrate and thiocyanate in different water samples. J. of Chromatographic Science 2012, 50(9), 826-830.
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