Pittcon 2016 (Complete)

Event Start Date:
March 6, 2016
Event End Date:
March 10, 2016
Event Venue:
Georgia World Congress Center in Atlanta

Visit us at Booth 3428!

Come see our Posters

Abstract Number: 750 – 9
Author Name: Ronald Benson – Shodex/Showa Denko America, Inc.
Session Title: Commercial Products Characterization
Event Type: Poster Sessions
Event Title: High Sensitive LC/MS Analysis of Stevia Sweeteners Using Polymer-Based Amino Column
Co-Author: Junji Sasuga, Satoko Sakai, Tomokazu Umezawa
Affiliation: Shodex/Showa Denko America, Inc.
Date: Monday, March 7, 2016, presenters present 1 pm to 3 pm
Start Time: (Slot #9)
Location: Exposition Floor, 400 Aisle


In recent years, such as Rebaudioside A, a glycoside derived from stevia, is added to a variety of processed foods as a sweetener for reduced calories. It is important to quantify the stevia sweeteners in processed food for quality control purposes. Herein we describe the highly sensitive and highly selective analytical separation by using polymer-based amino column with detection by LC/MS. Chromatographic separation was conducted using polymer-based amino column with isocratic mixture of (A) 0.1% NH3 aq. / (B) CH3CN as the eluent. The flow rate was 0.2mL/min and the temperature was 30 °C. ESI was selected for ionization method and SIM (-) for detection mode. Eight components, Rebaudioside A/B/C/D/F, Isosteviol, Stevioside, Steviolbioside, were mixed in each 50ng/mL as a standard sample. Injection volume was 5μL. Eight components were almost separated within 20 minutes and were detected with sufficient S/N in the condition of 78% CH3CN by HILIC mode. Rebaudioside B and Steviolbioside were strongly adsorbed to the column when the eluent (A) was only water since they have a carboxyl group. It became possible to adjust the appropriate retention time by alkaline with NH3. Result of measuring commercially available energy drink containing stevia sweeteners as a real sample, Rebaudioside A was detected, and additionally the other additives such as Sucralose, Acesulfame, Erythritol, Carnitine, Leucine, Isoleucine, Varine and Pyridoxine were also detected at the same time with good peak shape. The analysis condition show that the stevia sweetener in processed food and other high polarity food additives can be separated by HILIC mode and quantify with high sensitive.



Abstract Number: 1750 – 11
Author Name: Ronald Benson – Shodex/Showa Denko America, Inc.
Session Title: Liquid Chromatography
Event Type: Poster Sessions
Event Title: Investigation of Reverse-Phase – HILIC Continuous Analysis Using a One Column
Co-Author: Junji Sasuga, Tomokazu Umezawa, Hideyuki Kondo
Affiliation: Shodex/Showa Denko America, Inc.
Date: Wednesday, March 9, 2016, presenters present 10 am to noon
Start Time: (Slot #11)
Location: Exposition Floor, 400 Aisle


A sample matrix that includes both hydrophobic and hydrophilic substances are difficult to analyze by reversed phase chromatography (RPC) or hydrophilic interaction chromatography (HILIC) alone. The analysis of such matrices requires the separation using both an RPC column and a HILIC column. Herein, we describe the method for analyzing both hydrophobic and hydrophilic compounds simultaneously with one column by applying RPC/HILIC continuous analysis method.

First, the transition in the retention behavior of hydrophilic compounds (Cytosine, Uridine and Uracil) and hydrophobic compounds (Toluene and Naphthalene) corresponding to changing CH3CN ratio was examined by using CH3CN / H2O as eluent. Chromatographic separation was conducted using RPC column (4.6mmI.D. ×150mmL) packed with polyhydroxymethacrylate. As a result, the retention order was reversed in two conditions, 30% CH3CN and 90% CH3CN. It is suggested that the RPC mode acts in low CH3CN concentration and the HILIC mode act in high CH3CN concentration. By taking account of other conditions, it is confirmed that the RCP mode appears in under 50% CH3CN condition and HILIC mode appears in above 70% CH3CN condition.

Second, the analysis for a commercially available energy drink by LC/MS with gradient condition from RPC mode to HILIC mode continuously was attempted. As a result, the method that both hydrophobic and hydrophilic compound can be separated in a single measurement was established with twice sample injection. This method realizes quick equilibration as switching the mode and rapid analysis.

The polymer-based RPC column containing hydroxyl group which was considered this time has a unique performance that can be applied to HILIC mode as well. This column realizes efficient analysis for hydrophobic compound and hydrophilic compound in a single measurement.