Journal List > Transl Clin Pharmacol > v.23(2) > 1082613

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Shin, Meijering, Heuveln, Wieling, Halladay, Sahasranaman, and Hop: Validation of a method for the determination of thiocyanate in human plasma by UV/VIS spectrophotometry and application to a Phase I clinical trial of GDC-0425
Original Article

Translational and Clinical Pharmacology 2015;23(2):59-65.

Published online: 15 December 2015

DOI: https://doi.org/10.12793/tcp.2015.23.2.59

Validation of a method for the determination of thiocyanate in human plasma by UV/VIS spectrophotometry and application to a Phase I clinical trial of GDC-0425

Young G. Shin1, ∗, , Henri Meijering2, ∗, , Fred H. van Heuveln2, Jaap Wieling2, Jason Halladay1, Srikumar Sahasranaman1, Cornelis E.C.A. Hop1

1Genentech Inc., 1 DNA way, South San Francisco, CA 94080, USA

2QPS Netherlands B.V., Petrus Campersingel 123, 9713 AG Groningen, The Netherlands

∗Correspondence: Y. G. Shin; Tel: +82-42-821-5931, Fax: +82-42-821-5566, E-mail: yshin@cnu.ac.kr Current address: College of Pharmacy, Chungnam National University, Daejeon 35015, Republic of Korea

Y.G. Shin and H. Meijering contributed equally to this paper.

Received 10 August 201521 September 2015       Accepted 5 October 2015

Copyright © 2015 Korean Society for Clinical Pharmacology and Therapeutics

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The development and validation of a method for the determination of concentrations of thiocyanate in human plasma are described here. A modified colorimetric method of Bowler was used with the following alteration in Monica Manual, Part III. In order to obtain the same sensitivity in low amounts of clinical samples, quartz SUPRASIL® micro cuvettes have been used. The quantitation range was between 25–500 M. Accuracy and precision of the quality control samples, linearity of the calibration curve, dilution, spike recovery and stability under various conditions were evaluated in the validation of the method and all demonstrated acceptable results. All validation results met good laboratory practice acceptance and FDA requirements to be acceptable for application in clinical trials. The validated method has been used for a Phase I clinical study in cancer patients orally administered with either 60 mg or 80 mg of GDC-0425 containing a cyanide (CN-) group. The thiocyanate levels from patients before and after drug administration showed no clinically significant differences.

Keywords: Thiocyanate analysis, human plasma, GDC-0425, checkpoint kinase 1 inhibitor

References

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Figure 1.
Structure of GDC-0425.
tcp-23-59f1.tif
Figure 2.
Data from patients treated with GDC-0425 show that post-dose thiocyanate levels were comparable to baseline (pre-dose) levels∗.
tcp-23-59f2.tif
Table 1.
Calibration curve parameters for SCN−
  CALIBRATION CURVE CURVE PARAMETERS
  concentration (µM)  
Batch number 25.0 50.0 100 200 250 300 400 500 Slope Intercept Correlation coefficient
1 25.2 48.2 105 204 243 290 402 505 0.000826 0.00122 0.9993
2 25.6 47.2 101 203 246 304 403 501 0.00083 0.000777 0.9995
3 25.2 49.4 100 198 241 291 437 493 0.000827 –0.00183 0.9988
Table 2.
Reproducibility and repeatability (accuracy (%bias) and precision (CV%)) of QC samples
  ARTIFICIAL PLASMA K2-EDTA PLASMA
LLOQ QC Low QC Med QC High QC Low QC Med QC High
Theoretical concentration (µmol/L) 25.0 75.0 250 400 75.0 250 400
Reproducibility (n=3) %Bias 4.7 3.0 –1.0 –0.8 3.2 –0.2 1.4
  CV% 14.7 11.1 3.4 2.2 6.3 3.7 3.8
  Number 17 18 18 18 18 18 18
Repeatability (run 1) %Bias 11.1 10.3 1.5 0.0 2.0 –1.5 1.0
  CV%              
  Number 6 6 6 6 6 6 6
(run 2) %Bias 3.9 9.0 –1.3 0.7 7.6 4.3 5.9
  CV% 12.7 5.1 2.1 1.7 6.9 1.4 1.7
  Number 6 6 6 6 6 6 6
(run 3) %Bias –2.2 –10.2 –3.2 –3.0 0.0 –3.5 –2.6
  CV% 15.9 4.5 2.7 1.3 6.3 2.2 0.6
  Number 5 6 6 6 6 6 6
Table 3.
Stability tests of thiocyanate in artificial and K2-EDTA plasma and processed samples (n=3)
Stability Item Matrix Notes Mean Accuracy (%bias)
LLOQ QC Low QC Med QC High
Freeze-thaw –20oC K2-EDTA plasma 4 cycles – –3.5 – –4.8
  –70oC K2-EDTA plasma 4 cycles – –7.7 – –4.2
Short-term room temperature K2-EDTA plasma 24 hours – 16.9 – 3.4
  2–8oC$ K2-EDTA plasma 1 day – –15.1 – –3.7
    Artificial plasma 3 days 16.0 8.1 –3.7 –5.4
Long-term –20oC K2-EDTA plasma 31 days – 11.1 – 4.8
  –70oC K2-EDTA plasma 190 days – 6.2 – 2.7
  2–8oC Artificial plasma 4 days – 4.4 – 1.1
Process stability# dark K2-EDTA plasma 4 hours – –12.6 – –0.6
    Artificial plasma 4 hours – 8.1 – –0.5
Process stability# day light K2-EDTA plasma 2.5 hours – –3.9 – 0.8
    Artificial plasma 4 hours – –7.9 – –4.5

%bias with respect to nominal values; – = Not determined;

$ before storage at 2–8

o C sample were precipitated;

# results based on single injections at room temperature.

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