NCCLS H15-A3-2000 PDF

Full title and description

Reference and Selected Procedures for the Quantitative Determination of Hemoglobin in Blood; Approved Standard — Third Edition (NCCLS document H15-A3, December 1, 2000). This standard defines the reference hemiglobincyanide (HiCN / cyanmethemoglobin) method for measuring hemoglobin concentration in human whole blood, specifies composition and spectral characteristics of HiCN reference solutions, and gives calibration and photometer procedures required for assignment of values to control materials and for evaluation and calibration of automated hematology analyzers.

Abstract

H15-A3 (Third Edition) describes the HiCN cyanmethemoglobin reference measurement procedure for hemoglobin, including reagent composition and quality criteria, preparation and spectral characteristics of HiCN reference solutions, calibration of photometers, storage and stability of standards, and procedures needed for assigning values to control and calibration materials. The document is intended to support accurate hemoglobin determination for diagnostic use, instrument evaluation, and manufacturer calibration work.

General information

• Status: Approved standard — Third Edition (H15-A3); archived by the publisher but retained as technically valid and referenced for regulatory and laboratory use.
• Publication date: December 1, 2000.
• Publisher: National Committee for Clinical Laboratory Standards (NCCLS). The organization is now known as the Clinical and Laboratory Standards Institute (CLSI).
• ICS / categories: 11.100 — Laboratory medicine (Laboratory medicine / Hematology).
• Edition / version: Third Edition (H15-A3).
• Number of pages: 32 pages.

Scope

Part I of H15-A3 specifies the reference HiCN (cyanmethemoglobin) method for quantitative determination of hemoglobin concentration in human whole blood. The scope includes procedures and requirements necessary for whole-blood calibration of automated hematology analyzers, assignment of values to control and calibration materials, evaluation of instruments and alternative methods, and routine clinical laboratory applications for diagnostic purposes and monitoring therapy. The standard also provides specifications for HiCN reference solutions (composition, spectral characteristics, and storage/stability).

Key topics and requirements

• Principle and detailed procedure for the HiCN (cyanmethemoglobin) reference method for hemoglobin measurement.
• Composition, preparation, and spectral characteristics required for HiCN reference solutions and secondary standards.
• Calibration procedures for photometers and instrument calibration workflows tied to HiCN standards.
• Criteria and procedures for assigning values to control and calibration materials used in hemoglobin measurement.
• Quality control, performance specifications, and recommended acceptance criteria for method verification and instrument evaluation.
• Storage, stability, and handling recommendations for HiCN standards (including temperature and light considerations).
• Safety considerations related to cyanide-containing reagents and appropriate laboratory precautions.

Typical use and users

This standard is used by clinical laboratory personnel, reference laboratories, manufacturers of hematology analyzers and reagents, organizations that prepare or assign values to control/calibration materials, and regulatory bodies involved in IVD device evaluation. Typical uses include establishing traceable hemoglobin calibration, verifying analyzer performance, assigning values to quality control materials, and supporting method comparison or validation studies.

Related standards

Related documents and standards include the previous edition H15-A2 (superseded by H15-A3), CLSI/ NCCLS standards on internal quality control and analyzer verification (for example H26 series and other hematology guidance), and international documents addressing traceability and reference measurement systems for hemoglobin. Guidance from international hematology standardization bodies (such as ICSH) and metrology/traceability standards are also relevant when establishing reference material chains.

Keywords

Hemoglobin, HiCN, cyanmethemoglobin, hemiglobincyanide, reference method, calibration, photometer calibration, hematology analyzers, control material, reference standard, NCCLS, CLSI, laboratory medicine.

FAQ

Q: What is this standard?

A: H15-A3 is the Third Edition (approved standard) titled "Reference and Selected Procedures for the Quantitative Determination of Hemoglobin in Blood" published by NCCLS on December 1, 2000. It specifies the HiCN reference method and related reference materials and procedures for hemoglobin measurement.

Q: What does it cover?

A: It covers the cyanmethemoglobin (HiCN) reference procedure for measuring hemoglobin concentration, reagent and reference-solution composition and spectral characteristics, calibration and photometer procedures, storage and stability of standards, and procedures for assigning values to control and calibration materials and evaluating instruments.

Q: Who typically uses it?

A: Clinical laboratory scientists, reference laboratories, hematology instrument and reagent manufacturers, quality-assurance personnel, and regulatory reviewers use H15-A3 for calibration, method verification, instrument evaluation, and CV/control material value assignment.

Q: Is it current or superseded?

A: H15-A3 was published December 1, 2000. The document is identified by the publisher as an approved-level standard that is being retained in the collection but is archived (no longer under active consensus review) while remaining technically valid. Users should confirm with the Clinical and Laboratory Standards Institute (CLSI) for any later revisions or replacement documents; this statement reflects the document's status as of March 2, 2026.

Q: Is it part of a series?

A: Yes. H15-A3 is part of the NCCLS/CLSI series of hematology and laboratory medicine standards. It replaces the prior H15-A2 edition and is related to other CLSI documents addressing hematology analyzer performance, internal quality control, and method verification.

Q: What are the key keywords?

A: Hemoglobin, HiCN, cyanmethemoglobin, hemiglobincyanide, reference method, calibration, photometer, hematology analyzer, control material, NCCLS, CLSI.