A-069 Comparison of Three Hemoglobin A1c Methodologies in Variant and Non-Variant Samples Article Swipe

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Christopher L. Rosemark , Michael Nicklas , Christine L.H. Snozek , Baumann Nikola , Darci R Block ·

YOU? · · 2025 · Open Access · · DOI: https://doi.org/10.1093/clinchem/hvaf086.068

Background Hemoglobin A1c is used to assess glycemic control to both diagnose and monitor diabetes mellitus. Several different methodologies are available including high performance liquid chromatography (HPLC), capillary electrophoresis (CE), and immunoassay (IA). These methods are standardized and traceable to the Diabetes Control and Complications Trial (DCCT). The aim of this study was to evaluate the difference in HbA1c results between methods and percent of reportable results in variant samples. Methods The HPLC (Bio-Rad Laboratories, Inc; D-100), the CE(Sebia Inc; Capillarys 3), and IA(Roche Diagnostics Inc; Tina-quant Generation 3) were compared using two sets of residual patient samples left after physician ordered testing was complete: 1) samples with an absence of abnormal hemoglobin peaks (i.e. non-variant) detected by HPLC; 2) samples with abnormal hemoglobin peaks (i.e. variant) characterized by hemoglobin electrophoresis performed clinically or lacking an A1c peak by HPLC/CE. The samples were tested within manufacturers stability claims. 317 non-variant samples were compared between the HPLC and CE methods. The absolute difference was calculated for each method using this equation (HbA1c%avg – HbA1c%X) where HbA1c%avg is the average of the methods that produced reportable results and X is the result from each method evaluated. Differences less than or equal to +/-0.3% were considered acceptable. 28 of the 317 non-variant and 47 variant samples were tested by all 3 methodologies. The 47 variant samples grouped into 3 cohorts: common heterozygous variants (HbC, HbD, HbE, HbS and HbF, n=10), uncommon heterozygous variants (n=29) and homozygous variants lacking HbA (n=8). The absolute differences in variant sample results were calculated for samples that did not surpass manufacturer’s claims for producing an accurate result. The frequency of producing a numeric result in variant samples was calculated for each method. Results The HPLC/CE non-variant sample comparison (N=317) had results spanning the measuring range (3.8% - 14.0%) with average(range) of absolute difference=0.1(-0.4to0.6%) with 293/317(92.4%) samples within +/-0.3%. The HPLC/CE/IA non-variant sample comparison (N=28) had a result range=4.0%-9.4%, the average(range) of difference=0.1(-0.2to0.5%) with 25/28 (89.2%) of samples within +/-0.3%. Common heterozygous variant samples gave numeric results with no disqualifying flags in 9 of 10(HPLC), 8 of 10(CE), and 9 of 10(IA) samples tested. The average(range) difference from average=0.0(-0.3,0.5%) for HPLC, 0.0(-0.2,0.3%) for CE, and -0.1(-0.6,0.2%) for IA. Uncommon variant samples gave results in 4 of 29 (HPLC), 20 of 29 (CE), and 26 of 29 (IA) of samples tested. The average(range) difference from average=-0.1(-0.3,0.1%) for HPLC, 0.0(-1.0,1.1%) for CE, and 0.0(-1.1,1.0%) for IA. The homozygous variant (no HbA) samples gave results in 0 of 8 (HPLC), 0 of 8 (CE), and 4 of 8 (IA) of those tested. Conclusion All three methods meet their claims for accurately measuring hemoglobin A1c from non-variant and common hemoglobin variants. The advantage of HPLC and CE is the ability to identify hemoglobin variants that may impact HbA1c quantitation. The IA method provides the most numeric results. IA gave results even when HbA was absent. The lab should understand the capabilities and limitations of their HbA1c method on the population served.

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Type: article
Language: en
Landing Page: https://doi.org/10.1093/clinchem/hvaf086.068
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DOI: https://doi.org/10.1093/clinchem/hvaf086.068

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Title: A-069 Comparison of Three Hemoglobin A1c Methodologies in Variant and Non-Variant Samples

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Publication year: 2025

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Publication date: 2025-10-01

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Authors: Christopher L. Rosemark, Michael Nicklas, Christine L.H. Snozek, Baumann Nikola, Darci R Block

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Landing page: https://doi.org/10.1093/clinchem/hvaf086.068

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abstract_inverted_index.samples.	70
abstract_inverted_index.spanning	294
abstract_inverted_index.uncommon	238
abstract_inverted_index.variant)	127
abstract_inverted_index.variants	230, 240, 244, 461
abstract_inverted_index.10(HPLC),	346
abstract_inverted_index.HbA1c%avg	175
abstract_inverted_index.advantage	450
abstract_inverted_index.available	21
abstract_inverted_index.capillary	28
abstract_inverted_index.complete:	105
abstract_inverted_index.different	18
abstract_inverted_index.frequency	271
abstract_inverted_index.including	22
abstract_inverted_index.measuring	296, 440
abstract_inverted_index.mellitus.	16
abstract_inverted_index.performed	132
abstract_inverted_index.physician	101
abstract_inverted_index.producing	266, 273
abstract_inverted_index.stability	147
abstract_inverted_index.traceable	39
abstract_inverted_index.variants.	448
abstract_inverted_index.(HbA1c%avg	171
abstract_inverted_index.Background	1
abstract_inverted_index.Capillarys	81
abstract_inverted_index.Conclusion	431
abstract_inverted_index.Generation	88
abstract_inverted_index.HPLC/CE/IA	312
abstract_inverted_index.Hemoglobin	2
abstract_inverted_index.Tina-quant	87
abstract_inverted_index.accurately	439
abstract_inverted_index.calculated	164, 256, 281
abstract_inverted_index.clinically	133
abstract_inverted_index.comparison	290, 315
abstract_inverted_index.considered	203
abstract_inverted_index.difference	57, 162, 358, 394
abstract_inverted_index.evaluated.	194
abstract_inverted_index.hemoglobin	113, 124, 130, 441, 447, 460
abstract_inverted_index.homozygous	243, 407
abstract_inverted_index.population	497
abstract_inverted_index.reportable	66, 184
abstract_inverted_index.understand	486
abstract_inverted_index.Diagnostics	85
abstract_inverted_index.Differences	195
abstract_inverted_index.acceptable.	204
abstract_inverted_index.differences	250
abstract_inverted_index.immunoassay	32
abstract_inverted_index.limitations	490
abstract_inverted_index.non-variant	150, 209, 288, 313, 444
abstract_inverted_index.performance	24
abstract_inverted_index.capabilities	488
abstract_inverted_index.heterozygous	229, 239, 333
abstract_inverted_index.non-variant)	116
abstract_inverted_index.standardized	37
abstract_inverted_index.Complications	45
abstract_inverted_index.Laboratories,	75
abstract_inverted_index.characterized	128
abstract_inverted_index.disqualifying	341
abstract_inverted_index.manufacturers	146
abstract_inverted_index.methodologies	19
abstract_inverted_index.quantitation.	466
abstract_inverted_index.0.0(-0.2,0.3%)	363
abstract_inverted_index.0.0(-1.0,1.1%)	399
abstract_inverted_index.0.0(-1.1,1.0%)	403
abstract_inverted_index.293/317(92.4%)	307
abstract_inverted_index.average(range)	302, 322, 357, 393
abstract_inverted_index.chromatography	26
abstract_inverted_index.methodologies.	219
abstract_inverted_index.-0.1(-0.6,0.2%)	367
abstract_inverted_index.electrophoresis	29, 131
abstract_inverted_index.manufacturer’s	263
abstract_inverted_index.range=4.0%-9.4%,	320
abstract_inverted_index.average=0.0(-0.3,0.5%)	360
abstract_inverted_index.average=-0.1(-0.3,0.1%)	396
abstract_inverted_index.difference=0.1(-0.2to0.5%)	324
abstract_inverted_index.difference=0.1(-0.4to0.6%)	305
cited_by_percentile_year
countries_distinct_count	1
institutions_distinct_count	5
citation_normalized_percentile.value	0.62259432
citation_normalized_percentile.is_in_top_1_percent	False
citation_normalized_percentile.is_in_top_10_percent	False