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Stability of Morphine, Codeine, and 6-Acetylmorphine in Blood at Different Sampling and Storage Conditions

Author(s): Papoutsis I, Nikoloau P, Pistos C, Dona A, Stefanidou M, Spiliopoulou C, Athanaselis S

Publication: J Forensic Sci, 2014, Vol. 59(2), Page 550-4

Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of tube material, anticoagulant type, storage temperature and duration, and freeze-thaw cycling on levels of spiked morphine, codeine and 6-acetyl morphine in blood specimens. Pooled blood specimens were spiked with morphine and codeine or 6-acetyl-morphine. During freeze-thaw experiments, blood was thawed for 4 h at room temperature and then refrozen at -20 degrees C for 24 h. Specimens stored at 4 degrees C were assayed after 1 day, 1 week, 2 weeks and 1 month, but specimens stored at -20 degrees C were assayed after 1 week, 2 weeks, 1 month, 2 months and 3 months.

   Summary of Findings:

   After 1 month of storage of whole blood at 4 degrees C, morphine levels declined by 12-23%, codeine levels declined by 12-22%, and acetyl morphine levels declined by 13-95%. After 3 months of storage of whole blood at -20 degrees C, morphine levels declined by 18-32%, codeine levels declined by 7-19%, and acetyl morphine levels declined by 8-74%. Compared to initial levels, after 1 day at 4 degrees C, morphine and codeine concentrations were 5% higher in specimens stored in glass tubes and 5% lower in blood stored in polypropylene or polystyrene, regardless of anticoagulant. In contrast, after 1 day at 4 degrees C, acetyl-morphine levels were 1-20% lower than baseline, with higher levels observed in specimens anticoagulated with sodium oxalate than K2EDTA. After 1 week at -20 degrees C, levels of morphine and codeine were 2% higher to 6% lower, and levels of 6-acetyl-morphine 2-10% lower than initial levels, depending on tube material and anticoagulant type, but significance was not stated. After three freeze-thaw cycles, levels of morphine, codeine and 6-acetyl-morphine declined by 3.97%, 10.3%, and 20.8%, respectively, in whole blood specimens. Generally, the addition of NaF and storage at -20 degrees C instead of 4 degrees C increased the stability of morphine, codeine and 6-acetyl-morphine in spiked pooled whole blood. Further, storage of spiked blood in glass tubes, as opposed to polypropylene or polystyrene, generally increased the stability of morphine and codeine, and the use of oxalate instead of K2EDTA as an anticoagulant generally increased the stability of 6-acetyl-morphine.

   Biospecimens

   • Bodily Fluid - Blood

   Preservative Types

   • None (Fresh)
   • Other Preservative
   • Frozen

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   Small molecule	GC-MS

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage temperature	4 degrees C -20 degrees C
   Storage	Storage duration	1 day 1 week 2 weeks 1 month 2 months 3months
   Storage	Freeze/thaw cycling	0 cycles 3 cycles
   Storage	Type of storage container	Glass tube Polypropylene tube Polystyrene tube
   Biospecimen Preservation	Type of fixation/preservation	None (fresh) Frozen Refrigeration
   Biospecimen Acquisition	Anticoagulant	Sodium fluoride Sodium oxalate Potassium EDTA

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