Evaluation of factors influencing accuracy in the analysis of succinylacetone in dried blood spots.

Author(s): Peng M, Liu L, Peng L

Publication: Clin Chim Acta, 2012, Vol. 413, Page 1265-9

PubMed ID: 22522056 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of specimen volume, hematocrit level, and location of the DBS punch on LC-MS/MS quantification of spiked SUAC. Specified amounts of plasma were added back to washed cells (collected in citrate phosphate dextrose adenine anticoagulant) from a single donor to obtain the desired hematocrit level. These blood specimens were then spiked with SUAC, and different volumes were spotted on filter paper, dried overnight at room temperature, and stored in sealed bags with desiccant at -40 degrees C. The calibration and QC specimens (hematocrit 50%) were used for analysis the next day (no frozen storage).

   Summary of Findings:

   Increasing hematocrit values led to higher measured SUAC concentrations in DBS by LC-MS/MS, regardless of the original spiked SUAC concentration. Larger bias was observed as the difference in hematocrit level between the calibrator (50% hematocrit) and the specimen increased. For specimens spiked with 5.96 umol/L SUAC, those with a hematocrit level of 30% had a bias of -45% while specimens with hematocrit levels of 40%, 50%, and 60% had biases of -11.7%, 7.7%, and 23.8%. Measured SUAC concentration increased with increasing volumes of blood spotted into filter paper. With blood volumes of 70-100 uL, biases of 20-37% were observed compared to the 50 uL calibration specimens. Blood volumes of 10-50 uL resulted in biases of <13%. The authors state that punches taken from 5 different locations on the DBS had a coefficient of variation (CV) of 2.14% representing a negligible chromatographic effect.

   Biospecimens

   • Bodily Fluid - Blood

   Preservative Types

   • Other Preservative

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   Small molecule	LC-MS or LC-MS/MS

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Aliquots and Components	Aliquot size/volume	10 uL 30 uL 50 uL 70 uL 100 uL
   Biospecimen Aliquots and Components	Biospecimen components	30% hematocrit 40% hematocrit 50% hematocrit 60% hematocrit 5.99 umol/L SUAC 148.60 umol/L SUAC
   Biospecimen Aliquots and Components	Biospecimen heterogeneity	Dried blood spot sub-sampling (exact positions not specified)

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