Cancer Diagnosis Program | Biorepositories and Biospecimen Research Branch

help |

Optimal collection and storage conditions for catecholamine measurements in human plasma and urine.

Author(s): Boomsma F, Alberts G, van Eijk L, Man in 't Veld AJ, Schalekamp MA

Publication: Clin Chem, 1993, Vol. 39, Page 2503-8

PubMed ID: 8252722 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to examine the effects of delayed centrifugation, storage duration, and storage temperature on the three catecholamine (CA) concentrations in plasma and urine specimens. In addition, various preservative methods were investigated including the addition of glutathione to plasma and acidification or the addition of EDTA and sodium metabisulfite to urine.

Conclusion of Paper

The authors determined that optimal conditions for measurement of norepinephrine (NE), dopamine (DA), and epinephrine (E) concentrations in plasma specimens through HPLC include centrifugation within one hour of blood collection and storage of plasma for up to 1 month at -20 degrees C. Storage duration of plasma can be extended to 6 months with the addition of glutathione, or a year if storage is at -70 degrees C. The concentrations of DA, NE and E in urine specimens remain stable if stored at 4 degrees C for up to 1 month. The stability of urine DA, NE and E can be increased to 4 months by the addition of EDTA or sodium metabisulfite and to 1 year if acidified.

Studies

Study Purpose

The purpose of this study was to evaluate the effects of keeping blood at 4 or 20 degrees C for different amounts of time before centrifugation and holding prepared plasma at 4 or 20 degrees C for different amounts of time before freezing on CA concentration. The effect of preparing plasma using a refrigerated centrifuge was also determined.

Summary of Findings:

Concentrations of NE, E, and DA did not change significantly in plasma stored for at least 48 h at 4 degrees C or up to 24 h at 20 degrees C. However, NE and DA concentrations decreased (p<0.05) after plasma was left at 20 degrees C for 48 h. On the other hand, keeping blood at 20 degrees C prior to centrifugation for 2 hours or more significantly decreased the concentrations of NE and E, and after 4 hours, DA concentrations were also decreased (p<0.05). Keeping blood at 4 degrees C for 2 hours or more, prior to centrifugation, led to increased concentrations of NE (p<0.05), while E concentration increased after 3 hours at 4 degrees C (p<0.05). Results for DA were variable. Preparation of plasma using a refrigerated centrifuge had no effect on CA concentrations.

Biospecimens

Preservative Types

Frozen

Diagnoses:

Normal
Hypertension

Platform:

Analyte	Technology Platform
Small molecule	HPLC

Pre-analytical Factors:

Classification	Pre-analytical Factor	Value(s)
Biospecimen Aliquots and Components	Blood and blood products	Whole blood Plasma
Storage	Storage temperature	20 degrees C 4 degrees C -70 degrees C
Storage	Storage duration	0 h 0.5 h 1 h 2 h 3 h 4 h 24 h 48 h
Biospecimen Aliquots and Components	Centrifugation	Centrifugation delays investigated

Study Purpose

The purpose of this study was to determine the stability of NE, DA and E in heparin plasma specimens stored at various temperatures over time, with and without the addition of glutathione.

Summary of Findings:

NE, DA and E concentrations decreased after 1 day at 37 degrees C, 2-3 days at 20 degrees C, and after 1 week at 4 degrees C, with or without the addition of glutathione. At lower storage temperatures (-20 degrees C) glutathione kept NE, DA and E concentrations stable for up to 200 days while concentrations decreased after 1 month without glutathione. At -70 degrees C, NE, DA and E concentrations remained stable with or without the addition of glutathione.

Biospecimens

Bodily Fluid - Plasma

Preservative Types

Frozen
Other Preservative
None (Fresh)

Diagnoses:

Not specified

Platform:

Analyte	Technology Platform
Small molecule	HPLC

Pre-analytical Factors:

Classification	Pre-analytical Factor	Value(s)
Biospecimen Preservation	Type of fixation/preservation	None (fresh) Frozen Refrigeration
Storage	Storage temperature	37 degrees C 20 degrees C 4 degrees C -20 degrees C -70 degrees C
Storage	Storage duration	0 d to 365 d
Biospecimen Preservation	Fixative additive/buffer	Glutathione None

Study Purpose

The purpose of this study was to determine the stability of NE, DA and E in urine specimens stored at various temperatures over time. Urine specimens were either acidified (UA), had EDTA and sodium metabisulfite added (UB), or received no treatment (UO).

Summary of Findings:

Slight variations between NE, DA and E were seen, but in general, CA concentrations were stable for 1-2 days at 37 degrees C, and 4-8 days at 20 degrees C in both UO and UB. In UA, CA concentrations were stable for 3 weeks at 37 degrees C, and for at least 2 months at 20 degrees C. At 4 degrees C, CA concentrations were stable for 35 days in UO, up to 4 months in UB, and stayed relatively stable over the course of a year in UA. At -20 and -70 degrees C, all three hormones either remained stable or showed slight decreases over the course of a year in UO, UB, and UA.

Biospecimens

Bodily Fluid - Urine

Preservative Types

None (Fresh)
Frozen
Other Preservative

Diagnoses:

Normal

Platform:

Analyte	Technology Platform
Small molecule	HPLC

Pre-analytical Factors:

Classification	Pre-analytical Factor	Value(s)
Biospecimen Preservation	Type of fixation/preservation	Refrigeration Frozen None (fresh)
Storage	Storage temperature	37 degrees C 20 degrees C 4 degrees C -20 degrees C -70 degrees C
Storage	Storage duration	0 d to 365 d
Biospecimen Preservation	Fixative additive/buffer	EDTA and sodium metabisulfite Hydrochloric acid None

You Recently Viewed

News and Announcements

Most Downloaded SOPs in 2024

New Articles on the GTEx Project are Now FREELY Available!

Just Published!

More...