Glucose

Retaining polar ionizable and non-ionizable compounds is a challenging task in chromatography. This is due to the lack of retention and poor peak shape occuring due to secondary interactions on reversed-phase and HILIC columns. We have developed a method for the separation of glucose and lysine on the mixed-mode Amaze TH column. Glucose is a monosaccharide which is polar (hydrophilic) and non-ionizable. Lysine is one of the essential amino acids, it is polar and zwitterionic with two basic and one carboxylic groups. Glucose is retained separated by pure HILIC mechanism and lysine is retained by combination of HILIC and cation-exchange mechanisms. The retention time is controlled by the amount of ACN, buffer pH and buffer concentrations. Buffer pH affects the ionization state of lysine and the ionization state of the stationary phase. This method is robust. The Amaze TH column can be used to retain and separate amino acids, sugars, amino sugars, organic and inorganic acids as well as polar neutral compounds like sugars. The column is compatible with major detection technique (ELSD, CAD, RI, UV and mass spec)

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Conditions of Experiment

Column:	Amaze TH
Separation Modes:	HILIC, cation-exchnage
Column Dimenstions:	4.6x50 mm, 3 um, 100A
Mobile Phase:	65% ACN with 175 mM AmAc pH 5
Detection:	ELSD 45°C, RI-compatible
Sample:	1 mg/ml
Injection:	3 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Amino acid, Sugar, Supplement
Nature of compounds:	Hydrophilic, Neutral, Polar, Zwitterionic
Compounds:	Glucose, Lysine

Several techniques are used for the retention and separation of polar and polar ionizable compounds. The HILIC mixed-mode proves to be a powerful approach to independently control elution of ionizable compounds by changing buffer concentration and buffer pH. Glucose is a six member monosaccharide which is polar and neutral in nature. Gluconic acid is a polar and acidic sugar-based compound which is a product of glucose oxidation in nature. Gluconic acid naturally occurs in fruits, honey and wine. It is also used as a food additive to regulate acidity. Gluconic acid mostly exists in a linear form. Glucuronic acid is a cyclic sugar-based acid which is polar and acidic in nature. Glucuronic acid is used as a building block for proteoglycans and glycoglycerolipids. Glucuronic and gluconic acids are fermentation products in Kombucha tea. We have developed a HILIC/anion-exchange mixed-mode approach for the separation of neutral and acidic sugars.Sugars are retained by an HILIC mechanism and sugar-based acids are retained by HILIC and anion-exchange mechanisms. The retention time is controlled by the amount of ACN, buffer pH and buffer concentration. This method is robust and reproducible. It can be used for analysis of neutral acidic and basic sugar-like molecules in one run. This method is compatible with major detection techniques ( (ELSD, CAD, RI, UV and mass spec)

Amaze TH PURCHASE LEARN ABOUT Amaze TH

Conditions of Experiment

Column:	Amaze TH
Separation Modes:	HILIC, anion-exchnage
Column Dimenstions:	4.6x150 mm, 5 um, 100A
Mobile Phase:	ACN/water/AmAc Buffer
Detection:	ELSD, 50°C
Sample:	1 mg/ml
Injection:	3 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Organic acid, Sugar
Nature of compounds:	Acidic, Hydrophilic, Neutral, Polar
Compounds:	Glucose, Gluconic acid, Glucuronic acid

Several techniques are used for the retention and separation of polar and polar ionizable compounds. HILIC mixed-mode proves to be a powerful approach to independently control the elution of ionizable compounds by changing buffer concentration and buffer pH. Glucose is a six member monosaccharide which is polar and neutral in nature. Gluconic acid is polar and an acidic sugar-based compound which is a product of glucose oxidation in nature. Gluconic acid naturally occurs in fruits, honey and wine. It is also used as a food additive to regulate acidity. Gluconic acid mostly exists in a linear form. Gluconic acid is one of the fermentation products in Kombucha tea. We have developed a HILIC/anion-exchange mixed-mode approach for the separation of glucose and gluconic acid on the Amaze TH mixed-mode HPLC column. Glucose is retained by the HILIC mechanism and gluconic acid is retained by HILIC and anion-exchange mechanisms. The retention time is controlled by the amount of ACN, buffer pH and buffer concentration. The change in pH will affect ionization and the polarity of the stationary phase and ionizable analytes. This method is robust and reproducible. It can be used for the analysis of neutral acidic and basic sugar-like molecules in one run. This method is compatible with major detection techniques ( (ELSD, CAD, RI, UV and mass spec)

Amaze TH PURCHASE LEARN ABOUT Amaze TH

Conditions of Experiment

Column:	Amaze TH
Separation Modes:	HILIC, anion-exchnage
Column Dimenstions:	4.6x150 mm, 5 um, 100A
Mobile Phase:	ACN/water/AmAc Buffer
Detection:	ELSD, 50°C
Sample:	1 mg/ml
Injection:	3 uL
Flow rate:	1 ml/min

Analytes

Class of compounds:	Organic acid, Sugar
Nature of compounds:	Acidic, Hydrophilic, Neutral, Polar
Compounds:	Glucose, Gluconic acid