Analysis of Saccharin by HPLC

What is Saccharin？

Saccharin appears as odorless, white crystals or crystalline powder.
The pH of sodium salt aqueous solution is 6.0 – 7.5.
In nickle plating industries, saccharin is used as a brightener.
In food industries, it is used as a sweetener.

Fig.1　Structure of Saccharin

Applications of Saccharin

・Food industries
・Nickle plating industries
・Medication industries

Fig.2　Above: Chromatogram of of Saccharin in Standard sample, Below: Chromatogram of Blank sample 

Principle of HPLC

HPLC analysis is based on the affinity of the analyte toward both mobile phase and stationary phase. HPLC rapidly separates the components in a mixed sample as it pumps through stationary phase toward the detector.

HPLC analysis of Saccharin

Preparation of samples and standard for HPLC

1. Sample pretreatment e.g.dilutions, filtration
2. Calibration curve –
   Preparing the series of standard solution for calibration curve
3. Filling samples and standard into HPLC vials
4. Preparing eluent

Fig.3　Standard of Saccharin and Sample in HPLC vials

Preparation of HPLC

After preparation of Saccharin standard, HPLC start-up procedure is as follows.

1. Installation of suitable chromatography column
2. Conditioning of HPLC
3. Placing samples and standard solutions in the sample tray of HPLC
4. Creating the method & sequence file
5. Starting to run sequence as for method created

                   Fig.4　Chromatography column installation (Left ), HPLC instrument (Right)