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Designation: E3503 − 25 ´1Standard Test Method forDetermination of Hemicellulose in Herbaceous Biomass byHigh Performance Liquid Chromatography 1This standard is issued under the f i xed designation E3503; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon (´) indicates an editorial change since the last revision or reapproval.ε 1 NOTE—Patent information in Footnote 2 was updated editorially in July 2026.1. Scope1.1 This test method covers the quantif i cation of hemicel-lulose present in herbaceous biomass substrates using a diluteacid incubation period and high performance liquid chroma-tography (HPLC) analysis. This method can be utilized acrossa variety of substrates containing 1 % to 44 % hemicellulosiccontent. 21.2 This method intends to provide carbohydrate analysisfor evaluation of fermentation efficacy for various biomasssaccharides associated with hemicellulose. Current fermenta-tion technologies can convert arabinose, galactose, glucose,mannose, and xylose and will leave uronic acids and aceticacids unconverted, so they are not included in this assay. Itshould be noted that co-quantif i cation of xylose, mannose, andgalactose occurs. Additionally, glucose is simultaneously re-leased from starch, cellulose, and hemicellulose during thisassay and the source cannot be determined, so glucose is notquantif i ed in this assay.1.3 Units—The values stated in SI units are to be regardedas the standard. No other units of measurement are included inthis standard.1.4 This standard does not purport to address all of thesafety concerns, if any, associated with its use. It is theresponsibility of the user of this standard to establish appro-priate safety, health, and environmental practices and deter-mine the applicability of regulatory limitations prior to use.1.5 This international standard was developed in accor-dance with internationally recognized principles on standard-ization established in the Decision on Principles for theDevelopment of International Standards, Guides and Recom-mendations issued by the World Trade Organization TechnicalBarriers to Trade (TBT) Committee.2. Referenced Documents2.1 ASTM Standards: 3E1705 Terminology Relating to Bioenergy and IndustrialChemicals from BiomassE1756 Test Method for Determination of Total Solids inBiomassE1757 Practice for Preparation of Biomass for Composi-tional Analysis2.2 Other Documents:AOAC 935.29 Moisture in Malt – Gravimetric Method(AOAC 2005) 43. Terminology3.1 Def i nitions of Terms:3.1.1 For def i nitions of terms used in this standard, refer toTerminology E1705.N OTE 1—The user is advised that the def i nitions used by variousindustries, marketers, regulatory bodies, and otherASTM Committees candiffer from those in this test method. When using this test method, it iscritical to use terminology from Committee E48.3.1.2 arabinose, n—a f i ve-carbon monosaccharide commonin biomass.3.1.3 calibration verif i cation standard, CVS, n—standardsused in determining the quality of the calibration curve as wellas the quality of the standard reagents used in preparing thecalibration standards.3.1.4 f i ve-carbon monosaccharide, n—a simple sugar withf i ve carbon atoms also known as a pentose.1 This test method is under the jurisdiction of ASTM Committee E48 onBioenergy and Industrial Chemicals from Biomass and is the direct responsibility ofSubcommittee E48.05 on Biomass Conversion.Current edition approved Dec. 1, 2025. Published January 2026. DOI: 10.1520/E3503-25E01.2 The Determination of Hemicellulose in Herbaceous Biomass by High Perfor-mance Liquid Chromatography assay process and conditions are covered by a patentheld by Soliton, 5132 S Cliff Ave, Sioux Falls, SD 57108, https://www.solitonsciences.com/patents. Interested parties are invited to submit informa-tion regarding the identif i cation of an alternative(s) to this patented item to theASTM International Headquarters. Your comments will receive careful consider-ation at a meeting of the responsible technical committee which you may attend.3 For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at www.astm.org/contact. For Annual Book ofASTM Standards volume information, refer to the standard’s Document Summarypage on the ASTM website.4 Available from AOAC International, 2275 Research Blvd., Suite 300,Rockville, MD 20850-3250, http://www.aoac.org.Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United StatesThis international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for theDevelopment of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.1