Sucrose

HPLC Analysis of Xylitol, Sorbitol and Sucrose on Amaze HD Mixed-Mode Column
Application description

Sugars are polar non-ionic compounds. They are usually analyzed on HILIC columns. We have developed a new column and method for analysis of mono, di- and other saccharides by HPLC. Sugars are retained by a pure HILIC mechanism. The polarity of the stationary phase can be changed based on the pH of the mobile phase and buffer concentrations. The presence of salt is desired to increase the retention time of sugars. In general, monosaccharides will retain less tan di and trisaccharides. The Amaze HD column is compatible with 100% ACN and with 100% water, and can be used either in isocratic or wide gradient modes. This method is robust, reproducible, and allows the analyzation of all polar sugars and other polar compounds on one column.

Conditions of Experiment
Column: Amaze HD
Separation Modes: HILIC
Column Dimenstions: 4.6x50 mm, 5 um, 100A
Mobile Phase: 80% ACN with 20 mM AmFm pH 3
Detection: ELSD 40°C
Sample: 1 mg/ml
Injection: 3 uL
Flow rate: 1 ml/min
Analytes
Class of compounds: Sugar
Nature of compounds: Hydrophilic, Neutral, Polar
Compounds: Xylitol, Sorbitol, Sucrose
HPLC Analysis of Sugar Alcohols and Sugars on Amaze HD Mixed-Mode Column
Application description

Sugar alcohols and sugars are carbohydrates which are used in food and can be found in fruits, vegetables and other naturally occurring products. Simple sugars include monosaccharides like glucose, fructose and galactose. Sugar alcohols are derived from sugars and are occurring naturally, or synthesized by hydrogenation of sugars. Both sugars and sugar alcohols are polar with no chromophores. The most common approach for the analysis of sugars and sugar alcohols is HILIC chromatography. These hydrophilic compounds are retained by a polar mechanism on a polar stationary phase like Amaze HD. This column has a very polar group on the surface of silica gel which allows the use of less ACN for the retention of sugars and sugar alcohols. This method and the column can be used for the analysis of sugars and sugar alcohols in fruits, veggies, juices and other beverages. The Amaze HD is compatible with 100% ACN and 100% water. ELSD, CAD, RI or MS are required to retain and monitor sugars.

Conditions of Experiment
Column: Amaze HD
Separation Modes: HILIC
Column Dimenstions: 4.6x50 mm, 5 um, 100A
Mobile Phase: 80% ACN with 20 mM AmFm pH 3
Detection: ELSD 40°C
Sample: 1 mg/ml
Injection: 3 uL
Flow rate: 1 ml/min
Analytes
Class of compounds: Sugar
Nature of compounds: Hydrophilic, Neutral, Polar
Compounds: Xylitol, Mannitol, Sucrose, Maltose, Melezitose
Simultaneous HPLC Analysis of Sucrose, Fosfomycin and Common Counterions on Amaze TH Mixed-Mode Column
Application description

Sucrose, fosfomycin, sodium and TRIS are part of the fosfomycin formulation. Sucrose, fosfomycin, sodium and tris are very polar in nature. Fosfomycin is a broad-spectrum antibiotic which contains an epoxy fragment as well as phosphonic acid. This makes this drug very polar and acidic. Sodium ion and tromethamine are used as counter-ions for this polar drug molecule. All four compounds are different in nature - sucrose is polar and neutral, sodium is polar and basic, TRIS is polar and basic, and fosfomycin is polar and acidic. It is hard to retain and separate these compounds in RP chromatography. HILIC can retain these polar compounds and mixed-mode HILIC on the Amaze TH column allows to introduce cation- and anion-exchange mechanisms for better retention and better retention control. The presence of ionic interaction, in addition to HILIC, allows better control of elution and retention. The retention time can be controlled by three parameters - amount of acetonitrile, amount of buffer and the buffer pH. This method can be used for the analysis of drug formulation and level of fosfomycin in biofluids (serum, blood, urine, etc). The method is robust, reproducible and compatible with all major detection techniques.

Conditions of Experiment
Column: Amaze TH
Separation Modes: HILIC, cation-exchange, anion-exchange
Column Dimenstions: 4.6x150 mm 5 um, 100A
Mobile Phase: ACN/water/Buffer
Detection: ELSD
Sample: 1 mg/ml
Injection: 10 uL
Flow rate: 1 ml/min
Analytes
Class of compounds: Amines, Antibiotic, Drug, Inorganic cation, Organic acid, Sugar
Nature of compounds: Acidic, Basic, Hydrophilic, Neutral, Polar
Compounds: Fosfomycin, Sucrose, Sodium ion, Tromethamine
HPLC Analysis of Sugars, Amino Acids and Carboxylic Acids on Amaze TH Mixed-Mode Column
Application description

Mixed-mode stationary phases were designed to retain and separate compounds with drastic difference in properties using one column and method. Carboxylic acids, amino acids and sugars are poorly retained in reversed-phase chromatography. Derivatization procedures, ion-pairing reagents and HILIC columns can be used tp analyze these complex mixtures. The use of ion-pairing reagent makes MS detection problematic due to volatility issues of IP and signal suppression in MS. HILIC mixed-mode columns, like Amaze TH, have polar ionizable groups on the surface which help significantly increase retention time of polar ionizable analytes like amines, amino acids, carboxylic acids and sugars. We have developed a method for separation and retention of succinic acid, phenylalanine, sucrose, glycine, aspartic acid and raffinose. Isocratic conditions provide separation within 15 minutes with good peak shape and retention control. Retention time on Amaze TH column is controlled by amount of ACN, buffer concentration, buffer ph and buffer nature. Method can be used for other amines, amino acids, carboxylic acids and sugars where fast MS-compatible separation is required.

Conditions of Experiment
Column: Amaze TH
Separation Modes: HILIC, cation-exchange, anion-exchange
Column Dimenstions: 4.6 x 100 mm 5 um, 100A
Mobile Phase: 80% ACN with 10 mM AmAc pH 4.8
Detection: ELSD
Sample: 0.3 mg/ml
Injection: 3 uL
Flow rate: 1 ml/min
Analytes
Class of compounds: Amino acid, Organic acid, Sugar, Supplement
Nature of compounds: Acidic, Hydrophilic, Neutral, Polar, Zwitterionic
Compounds: Succinic acid, Phenylalanine, Sucrose, Glycine, Aspartic acid, Raffinose