Abstracts
349
Phenobarbital assay evaluation on the Abbott Architect:
Something old, something new
Daniel Beriaulta, Paul Martensa, Paul Yipb
a
University of Toronto, Toronto, ON, Canada
b
University Health Network, Toronto, ON, Canada
Objective: To evaluate the analytical performance of the recently
released Abbott Architect Phenobarbital PETINIA immunoassay (LN
5P07), and compare it to the previous Architect enzyme immuno-
assay (LN 1E08) on the Architect c8000 chemistry analyzer.
Methods: Precision was assessed using Bio-Rad Immunoassay
Plus quality control (QC) material. Analysis was performed over
10 days with 2 runs per day in duplicate using a single calibration.
The previous phenobarbital calibrator has been replaced with a new
multi-constituent calibrator for several TDM assays. The benefits of
the new calibrator include longer onboard and shelf-life stability.
Linearity was assessed with CAP survey specimens. Limit of blank
study was performed with 20 replicates of normal saline. Inter-
method comparison was assessed using plasma specimens and a
single replicate for each assay.
Results: Phenobarbital imprecision was determined at three QC
levels with a quality goal of 5%. Levels 1–3 spanned a range of 41.2–
220.7 μmol/L with CV's of 2.7–3.2%. The linear range was verified
over 42.1–295.1 μmol/L and the upper analytical measurement range
without dilution was 344.8 μmol/L. The verified limit of blank was
1.05 umol/L, with a stated limit of quantitation of 8.6 umol/L. Inter-
method comparison included 50 patient samples over a range of 0.3–
261.1 μmol/L and the regression gave values with slope = 0.895,
intercept = 11.42, and R = 0.977.
Conclusions: Evaluation of the analytical performance of the
Phenobarbital PETINIA assay met the manufacturer’s claims and was
deemed acceptable for patient testing.
Keywords: Phenobarbital, Method Evaluation, Abbott Architect,
Immunoassay, Drug Monitoring
doi:10.1016/j.clinbiochem.2015.07.092
350
Online sample preparation integrated with LC/MS–MS for routine
use in a clinical laboratory
Michel Dery, Dominique Guérette
CHU de Québec-Université Laval, Québec, QC, Canada
Objective: In the past decade, liquid chromatography coupled with
tandem mass spectrometry (LC/MS–MS) has transformed the way many
compounds are measured in clinical laboratories. LC/MS–MS is often
used to measure analytes that are difficult to measure by immunoassay.
The goal of this study is to show that online sample preparation using
solid phase extraction (SPE) provides a robust method for the analysis of
immunosuppressants and steroid compounds.
Design and methods: This study details two methods using
deuterated internal standards and commercial calibrators
(Chromsystems) based on an LC/MS–MS (ACQUITY/XEVO MS:
Waters) coupled with the online SPE Manager [OSM] (Cartridges
XBridge C18) from Waters. Protein precipitation using zinc sulfate
was performed prior to the injection of the samples onto the OSM.
1025
Results: Whole blood samples treated with the OSM are 5–6
times less affected by the matrix effects seen in untreated samples.
This improvement in sensitivity and signal led to a significant
improvement to the assay performance. When compared to UK-
NEQAS results (30 samples), a bias of −4% (SD: 7) was obtained for
sirolimus measured with the OSM, while a bias of +7% (SD: 30)
without the OSM was observed. Furthermore, a method also using
OSM was able to accurately and reproducibly measure testosterone
levels in women and infants.
Conclusions: These results show that by automating sample
preparation for LC/MS–MS, a number of sensitive, accurate, and
highly efficient tests using the full analytical power of LC/MS–MS can
be developed and deployed in a clinical laboratory.
doi:10.1016/j.clinbiochem.2015.07.093
351
Liaison XL analyzer evaluation of three Diasorin assays —
insulin-like growth factor 1 (IGF1), 25-hydroxyvitamin D (25D)
and the recently released fully automated “no prep”
1,25-dihydroxyvitamin D (1,25D)
Daniel Beriaulta, Michael Knauerb, Barry Hoffmanc
a
University of Toronto, Toronto, ON, Canada
b
Department of Laboratory Medicine and Pathobiology, University of
Toronto, Toronto, ON, Canada
c
Mount Sinai Hospital, Toronto, ON, Canada
Objective: Evaluate the analytical performance of three fully
automated DiaSorin immunoassays (IGF1, 25D and 1,25D) run on the
Liaison XL analyzer. The 1,25D method directly quantifies serum/
plasma specimens without the need for pre-analytical processing
designed to remove cross-reactants and disrupt protein binding.
Methods: Precision, linearity and inter-method comparison were
assessed according to modified CLSI protocols (EP5-A2, EP6-A, and
EP9-A3, respectively). DEQAS-PT specimens were analyzed for 1,25D
and results were compared to the mass spectrometry mean reported
by DEQAS.
Results: IGF1, 25D and 1,25D imprecision determined across ≥2
reagent lots, multiple calibrations and 7–14 runs of 2–5 replicates
each varied, respectively, from 4-8% (60-410 μg/L), 4.6–7.3% (39-
125 nM) and 7.3–12.6% (60–270 pM). Serum pools with elevated
IGF1, 25D or 1,25D (710 μg/L, 155 nM, 515 pM, respectively) were
diluted with assay diluent (IGF1) or low concentration serum (25D,
1,25D; saline unacceptable) and the diluted series were assessed for
linearity. R2 exceeded 0.99 and recovery was within ±20% of
expected. DiaSorin 25D assay compared against mass spectrometry
(55 patient specimens, range 20–150 nM) yielded DiaSorin 25D =
0.82xMass Spectrometry + 12 (R2 = 0.87). DiaSorin IGF1 compared
against IDS iSYS (40 patient specimens, range 26–600 μg/L) yielded
DiaSorin IGF1 = 1.07xIDS iSYS + 13 (R2 = 0.99). DiaSorin 1,25D
recovered within ±10% of the DEQAS mass spectrometry mean for
the 10 DEQAS specimens tested on the Liaison XL.
Conclusions: Evaluation of the analytical performance of the three
assays met the manufacturer's claims. The DiaSorin 25D and 1,25D
assays agreed reasonably well with mass spectrometry analysis.
doi:10.1016/j.clinbiochem.2015.07.094