HPLC Method for Analysis of Chlorhexidine in Biofluids: Blood Plasma and Urine on BIST B+ Column

HPLC Method for Analysis of Chlorhexidine Biofluids: Blood Plasma and Urine on BIST B+ by SIELC Technologies.

Separation type: Bridge Ion Separation Technology, or BIST™ by SIELC Technologies

HPLC Method for Analysis of Chlorhexidine in Biofluids Blood Plasma and Urine on BIST B+ Column
HPLC Method for Analysis of Chlorhexidine on BIST B+ by SIELC Technologies.

High Performance Liquid Chromatography (HPLC) Method for Analyses of Chlorhexidine

Chlorhexidine is a common antiseptic used intensively in hand sanitisers and other antibacterial products. Typical HPLC methods used to analyze this product are done in reverse phase (RP) mode, but this is complicated by the presence of two basic groups which, with the overall hydrophobic characteristics of the molecule, usually produce asymmetrical peaks.


ColumnBIST B+, 4.6×150 mm, 5 µm, 100A
Mobile PhaseMeCN
BufferH2SO4 – 0.2%
Flow Rate1.0 ml/min
DetectionUV 260 nm
Peak Retention Time6.2 min


Class of CompoundsTopical antiseptic
Analyzing CompoundsChlorhexidine

Sample preparation:

Plasma/urine sample was spiked with a small amount of Chlorhexidine and then diluted 5 times with acetonitrile. A formed precipitate was removed by centrifugation and the cleared solution was used directly for injection. 

Application Column


BIST™ columns offer a unique and effective way to achieve separations that were traditionally challenging or even impossible with other HPLC columns. With the use of a special mobile phase, these ion exchange columns provide very strong retention for analytes with the same charge polarity as the stationary phase, unlocking new chromatography applications. What makes BIST™ columns stand out is their proprietary surface chemistry, which results in superior selectivity, resolution, and sensitivity. These columns offer a simple, efficient solution for a variety of analytical challenges, making them an excellent choice for researchers and analysts across many different fields. To learn more about the technology that powers BIST™ columns and to explore related applications, check out https://BIST.LC.

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Application Analytes:
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.