Honey Sugars by HPLC-CAD

Corona TM Charged Aerosol DetectorHoney SugarsApplication noteThe need for universal HPLC detection inanalytical laboratories is widespread.While several detection technologies(e.g., low wavelength UV, refractive index,evaporative light scattering,chemiluminescent nitrogen detectors) arecurrently being used, there is significantroom for improvement in performancecharacteristics such as sensitivity, dynamicrange, consistency of response factors andgradient or solvent compatibility.To help address the many challenges ofuniversal detection, ESA has developed theCorona CAD TM detector. This noveltechnology offers many benefits toanalytical scientists including:• High Sensitivity - Low ng limits ofdetection.• More Consistent Response Factors -Response magnitude does not significantlydepend on analyte properties (e.g. molarabsorbtivity, proton affinity).• Broad and Useful Dynamic Range -4 orders of magnitude (ng - µg quantities).• Excellent Reproducibility - Typically lessthan 2% RSD.• Broad Applicability - Can be used with awide variety of HPLC conditions tomeasure virtually any nonvolatile analyteincluding proteins, lipids, carbohydratesand small molecules.• Ease of Use - Easy setup. Uses minimalbench space and requires only gas inputpressure and signal output range to be set.Figure 1. Analysis of Eight Carbohydrate Standards (1µg each oncolumn). Turanose and Saccharose could not be resolved.This application note describes the use of the Corona CAD forthe measurement of a number of sugars typically found inhoney (Figure 1). The method had a limit of detection of ~10ng(on column), showed good precision and had a dynamic rangefrom ng to µg levels (Figure 2). The method can be used todetermine sugar abundance in different varieties of honey(Figure 3). This is an example of detection of analytes that onlypossess weak chromophores.Corona parametersGas: 35psi via nitrogen generatorFilter: noneRange: 100pAHPLC Parameters:Mobile Phase: 70% (v/v) aqueous acetonitrileFlow Rate: 1.0mL/minColumn: Shodex Asahipak, NH2P-50 4E, 4.6 x 250mm; 5µmColumn Temperature: 35 o CInjection Volume: 10µL

The CoronaTMCharged Aerosol DetectorResponse200000150000100000Raffinose (r2 = 0.9998)Melezitose (r2 = 0.9999)Maltose (r2 = 0.9995)Trehalose (r2 = 0.9999)Glucose (r2 = 0.9999)Fructose (r2 = 0.9999)Sample preparationStandards were obtained from Sigma (St. Louis, MO)and dissolved in water (1mg/mL). Further dilutionswere in water. Honey samples were diluted in waterprior to analysis.Conclusions5000000 200 400 600 800 1000Mass (ng on column)Figure 2. Response Curves.The Corona CAD provides universal detection ofnonvolatile analytes with response independent ofchemical properties, a wide dynamic response range,high sensitivity and good precision. Thesecharacteristics, along with reliability and simpleoperation, make this a superior detector for a widerange of HPLC analyses.For more information about this application, theCorona CAD, or charged aerosol detection visitwww.coronacad.com. We are interested in youropinions and are available to answer any questions youmay have: please contact a technical representative at978.250.7082, or if e-mail is more convenient, sendyour questions to coronacad@esainc.com.Ordering informationFigure 3. Analysis of Different Diluted Honey Samples.DescriptionCoronaThermal Organizer ModuleNitrogen generator 70-6003Pump, model 582 70-4050Autosampler, model 542 70-4152Elite software including PC 70-5073Column, Shodex Asahipak, NH2P-50 4EPart Number70-6350 (100/120V)70-6351 (230/240V)70-5499TAESA Biosciences Inc. • 22 Alpha RoadChelmsford, MA 01824-4171U.S.A.Tel: (978) 250-7000Fax: (978) 250-7090www.esainc.comwww.coronacad.comESA Analytical • Brook FarmDorton, AylesburyBuckinghamshireHP18 9NHEnglandTel: 44 (0)1844 239381www.esainc.comESA, Inc. is an ISO 9001 and 13485 Certified CompanyCorona TM and CAD TM are trademarks of ESA, Inc.ESA ®is a registered trademark of ESA, Inc.The Corona CAD is covered by multiple patents70-6977IA-1