Glycerol at Low Levels by HPLC-CAD

Corona TM Charged Aerosol DetectorGlycerol at low levelsApplication noteThe need for universal HPLC detection inanalytical laboratories is widespread.While several detection technologies(e.g., low wavelength UV, refractive index,evaporative light scattering, chemiluminescentnitrogen detectors) are currentlybeing used, there is significant room forimprovement in performance characteristicssuch as sensitivity, dynamic range,consistency of response factors andgradient or solvent compatibility.To help address the many challenges ofuniversal detection, ESA has developedthe Corona CAD TM detector. This noveltechnology offers many benefits toanalytical scientists including:• High Sensitivity - Low ng limits ofdetection.Response2625.52524.5241 2 3 4 5MinutesFigure 1. Glycerol ~1.26ng (on column) in mobile phase.Glycerol• Consistent Response Factors - Responsemagnitude does not significantly dependon analyte properties (e.g. molarabsorbtivity, proton affinity).• Broad and Useful Dynamic Range - 4orders of magnitude (ng - µg quantities).• Excellent Reproducibility - Typically lessthan 2% RSD.• Broad Applicability - Can be used witha wide 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 beset.This application note describes the use of the Corona CAD forthe measurement of glycerol. The method has excellentsensitivity (Figure 1), precision (Figure 2) and a dynamic rangethat covers ng to µg levels. This is an example of detection ofan analyte that lacks a chromophore.Corona parametersGas: 35psi via nitrogen generatorFilter: noneRange: 100pAHPLC Parameters:Mobile Phase: 75% (v/v) acetonitrileFlow Rate: 1.0mL/minColumn: Shodex Asahipak NH2-50 4E 4.6 x 250mmColumn Temperature: ambientInjection Volume: 10µLSample preparationStock solution (Sigma - G5516) was diluted in water.

The CoronaTMCharged Aerosol DetectorConclusionsResponse700600500400300GlycerolThe Corona CAD provides universal detection of nonvolatileanalytes 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.20010001 2 3 4 5MinutesFigure 2. Reproducibility (5 replicates of 126µg standard).For more information about this application, theCorona CAD, or charged aerosol detection visitwww.coronacad.com. We are interested in your opinionsand 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 informationDescriptionCoronaThermal Organizer ModuleNitrogen generator 70-6003Pump, model 582 70-4050Autosampler, model 542 70-4152Quaternary low pressure gradientand degasser 70-5260Elite software including PC 70-5073Shodex Asahipak NH2-50 4EPart Number70-6350 (100/120V)70-6351 (230/240V)70-5499TAESA Biosciences, Inc.22 Alpha RoadChelmsford, MA 01824-4171 USA(978) 250-7000 / Sales(800) 959-5095Fax (978) 250-7090www.esainc.com /www.coronacad.comESA Analytical • Brook FarmDorton, AylesburyBuckinghamshireHP18 9NHEnglandTel: 44 (0)1844 239381www.esainc.comESA Biosciences, Inc. A Magellan Biosciences CompanyESA, 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 patents70-6691IA-1