Standard PREanalytical Code Version 4.0

Abstract

Conformity to preestablished standard operating procedures (SOPs) is crucial for the consistency of biospecimen quality and thus for the success of research using biospecimens.^1–4 Standardized terminology and reporting of biospecimen provenance and processing can improve the robustness and reproducibility of analytical results. Preanalytical variables play an integral role in the outcome of the research and thus quality assurance focus has become a priority.^5,6 The continuous evolution of scientific technologies and dynamic research requirements for precision medicine has necessitated the development of a foundational framework for biospecimen traceability and its periodic updates.⁷ The Standard Preanalytical Code (SPREC) is a seven-element code that facilitates the annotation of biospecimens by identifying preanalytical factors that significantly influence downstream analyses and impact analysis outcomes, allowing these factors to be anticipated and standardized within the SOPs. SPREC offers overarching guidance and a general framework for the documentation and reporting of preanalytical variables, applicable across various research contexts. Support tools for SPREC implementation in biobank IT systems have been previously described.⁸ Specific operational requirements, especially those related to complex derivatives production, should be considered and incorporated into their respective laboratory protocols or SOPs. This would ensure complete, detailed, and accurate documentation and handling of all preanalytical variables, aligned with the unique needs of each study or clinical investigation. Updates to the SPREC were done in 2012 and 2018^8,9 and both the ISBER Best Practices¹⁰ and the ISO20387:2018¹¹ make reference to the SPREC.

In this short communication, the SPREC version 4.0 is published. It includes updates of new sample types and types of corresponding primary containers, additional options in sample storage and processing times and temperatures during the first and second centrifugation, and finally more options for longer tissue fixation times, based on feedback and real-time experiences from SPREC implementation in biobanking workflows (Tables 1 and 2). SPREC options are never removed in more recent versions, so that SPREC data that have been previously documented in biobank databases always remain valid.

Table 1.

Preanalytical Variables, with New Options in Bold Italic, Included in Standard Preanalytical Code (7-Element Long SPREC) Version SPREC 4.0, Applied to Fluid Samples

SPREC preanalytical variables and options		Code
Type of sample
Ascites fluid		ASC
Amniotic fluid		AMN
Bronchoalveolar lavage		BAL
Blood (whole)		BLD
Bone marrow aspirate		BMA
Breast milk		BMK
Buccal cells		BUC
Nondensity-gradient-centrifugation-separated buffy coat, viable		BUF
Nondensity-gradient-centrifugation-separated buffy coat, nonviable		BFF
Density-gradient-centrifugation-separated mononuclear cells, viable		CEL
Fresh cells from nonblood specimen type		CEN
Cells from nonblood specimen type (e.g., ascites, amniotic), viable		CLN
Cord blood		CRD
Cerebrospinal fluid		CSF
Enriched (physicochemically) circulating tumor cells		CTC
Exhaled breath condensate		EBC
Follicular fluid		FLF
Dried whole blood (e.g., Guthrie cards)		DWB
Nasal washing		NAS
Density-gradient-centrifugation-separated mononuclear cells, nonviable		PEL
Isolated or enriched exosomes		EXO
Cells from nonblood specimen type (e.g., ascites, amniotic), nonviable		PEN
Pleural fluid		PFL
Dental pulp		PLP
Plasma, single spun		PL1
Peritoneal fluid		PTF
Plasma, double spun		PL2
Red blood cells		RBC
Saliva		SAL
Semen		SEM
Serum		SER
Sputum		SPT
Stool		STL
Sweat		SWT
Synovial fluid		SYN
Tears		TER
24 h urine		U24
Urine, random (“spot”)		URN
Urine, first morning		URM
Urine, timed		URT
Other		ZZZ
Type of primary container
Acid citrate dextrose		ACD
Active cooling device for condensate		ACL
Serum separator tube without clot activator		CAT
Citrate phosphate dextrose		CPD
Citrate fluoride		CIF
Cell Preparation Tube^® citrate		CPT
Cell Preparation Tube heparin		CPH
Aldehyde-based stabilizer for CTCs		CSV
EDTA and gel		EDG
Physical filtration system		FIL
Glass		GLS
Lithium heparin		HEP
Hirudin		HIR
Lithium heparin and rubber plug		LHB
Lithium heparin and gel		LHG
Oragene collection container or equivalent		ORG
Stool collection container with DNA stabilizer		OMN
Passive cooling device for condensate		PCL
PAXgene^® blood RNA⁺		PAX
Potassium EDTA		PED
Polyethylene tube sterile		PET
S8820 protease inhibitor tablets or equivalent		PI1
Protease inhibitors		PIX
Polypropylene tube sterile		PPS
PAXgene blood DNA		PXD
PAXgene bone marrow RNA		PXR
Sodium citrate		SCI
Nonaldehyde-based stabilizer for cell-free nucleic acids		SCK
Sodium EDTA		SED
Sodium heparin		SHP
Sodium fluoride/potassium oxalate		SPO
Serum separator tube with clot activator		SST
Tempus^® tube		TEM
Trace elements tube		TRC
Unknown		XXX
Other		ZZZ
Precentrifugation (delay between collection and processing)
RT	<30 min	A1
2°C–10°C	<30 min	B1
RT	<2 h	A
2°C–10°C	<2 h	B
RT	2–4 h	C
2°C–10°C	2–4 h	D
RT	4–8 h	E
2°C–10°C	4–8 h	F
RT	8–12 h	G
2°C–10°C	8–12 h	H
RT	12–24 h	I
2°C–10°C	12–24 h	J
RT	24–48 h	K
2°C–10°C	24–48 h	L
RT	>48 h	M
2°C–10°C	>48 h	N
>35°C	<2 h	O
Unknown		X
Other		Z
Centrifugation
RT 10–15 min	<3000 g no braking	A
RT 10–15 min	<3000 g with braking	B
2°C–10°C 10–15 min	<3000 g no braking	C
2°C–10°C 10–15 min	<3000 g with braking	D
RT 10–15 min	3000–6000 g with braking	E
2°C–10°C 10–15 min	3000–6000 g with braking	F
RT 10–15 min	6000–10,000 g with braking	G
2°C–10°C 10–15 min	6000–10,000 g with braking	H
RT 10–15 min	>10,000 g with braking	I
2°C–10°C 10–15 min	>10,000 g with braking	J
RT 30 min	<1000 g no braking	M
RT <10 min	3000–6000 × g	O
2°C–10°C >15 min	<3000 g	P
2°C–10°C >15 min	<3000 g with braking	Q
2°C–10°C 30–120 min	>10,000 g with braking	R
2°C–10°C	>80,000 g	U
No centrifugation		N
Unknown		X
Other		Z
Second centrifugation
RT 10–15 min	<3000 g no braking	A
RT 10–15 min	<3000 g with braking	B
2°C–10°C 10–15 min	<3000 g no braking	C
2°C–10°C 10–15 min	<3000 g with braking	D
RT 10–15 min	3000–6000 g with braking	E
2°C–10°C 10–15 min	3000–6000 g with braking	F
RT 10–15 min	6000–10,000 g with braking	G
2°C–10°C 10–15 min	6000–10,000 g with braking	H
RT 10–15 min	>10,000 g with braking	I
2°C–10°C 10–15 min	>10,000 g with braking	J
RT 15–30 min	3000–6000 g with braking	K
RT 15–30 min	3000–6000 g without braking	L
RT 30–60 min	3000–6000 g	M
2°C–10°C	>80,000 g	U
No centrifugation		N
Unknown		X
Other		Z
Postcentrifugation delay
<1 h 2°C–10°C		A
<1 h RT		B
1–2 h 2°C–10°C		C
1–2 h RT		D
2–8 h 2°C–10°C		E
2–8 h RT		F
8–24 h 2°C–10°C		G
8–24 h RT		H
24–48 h 2°C–10°C		I
24–48 h RT		J
>48 h RT		M
Not applicable		N
Unknown		X
Other		Z
Long-term storage
PP tube 0.5–2 mL	(−85) to (−60)°C	A
PP tube 0.5–2 mL	(−35) to (−18)°C	B
PP tube 0.5–2 mL	<−135°Ca	V
Cryotube^b 1–2 mL	LN	C
Cryotube^b 1–2 mL	(−85) to (−60)°C	D
Cryotube^b 1–2 mL	Programmable freezing to <−135°C	E
Plastic cryo straw	LN	F
Straw	(−85) to (−60)°C	G
Straw	(−35) to (−18)°C	H
Straw	Programmable freezing to <−135°C	I
PP tube ≥5 mL	(−85) to (−60)°C	J
PP tube ≥5 mL	(−35) to (−18)°C	K
Microplate well	(−85) to (−60)°C	L
Microplate well	(−35) to (−18)°C	M
Cryotube^b 1–2 mL	LN after temporary (−85) to (−60)°C	N
Plastic cryo straw	LN after temporary (−85) to (−60)°C	O
Paraffin block	RT or 2°C–10°C	P
Paraffin block	(−35) to (−18)°C	U
Bag	LN	Q
Dry technology medium	RT	R
PP tube 40–500 μL	(−85) to (−60)°C	S
PP tube 40–500 μL	(−35) to (−18)°C	T
PP tube 40–500 μL	<−135°Ca	W
Original primary container	(−35) to (−18)°C or (−85) to (−60)°C	Y
Unknown		X
Other		Z

Codes in bold come from the LDMS. Volumes refer to container size.

Temperature <−135°C may correspond to LN vapor phase or to −150°C electrical freezer.

Cryotube is defined as a tube that can be stored in LN either vapor or liquid phase.

h, hour; LDMS, Laboratory Data Management System; LN, liquid nitrogen, referring to either vapor or liquid phase (this specific information should be documented in the biobank’s SOPs); min, minute; PP, polypropylene; RT, room temperature: 18°C–28°C; SOPs, standard operating procedures; SPREC, Standard Preanalytical Code.

Table 2.

Preanalytical Variables, with New Options in Bold Italic, Included in the Standard Preanalytical Code (7-Element Long SPREC) Version SPREC 4.0, Applied to Solid Samples

SPREC preanalytical variables and options		Code
Type of sample
Bone		BON
Fresh cells from nonblood specimen type (e.g., biopsy)		CEN
Cells from nonblood specimen type (e.g., dissociated tissue), viable		CLN
Cells from fine needle aspirate		FNA
Hair		HAR
Cells from laser capture microdissected tissue		LCM
Nails		NAL
Cells from nonblood specimen type (e.g., dissociated tissue), nonviable		PEN
Placenta		PLC
Solid tissue		TIS
Disrupted tissue, nonviable		TCM
Teeth		TTH
Other		ZZZ
Type of collection
Autopsy <6 h postmortem		A06
Autopsy 6–12 h postmortem		A12
Autopsy 12–24 h postmortem		A24
Autopsy 24–48 h postmortem		A48
Autopsy 48–72 h postmortem		A72
Biopsy in culture media		BCM
Biopsy		BPS
Biopsy in normal saline or phosphate buffered saline		BSL
Biopsy in tissue low-temperature transport media		BTM
Fine needle aspirate		FNA
Punction		PUN
Surgical excision in culture media		SCM
Surgical excision		SRG
Surgical excision in normal saline or phosphate buffered saline		SSL
Surgical excision in tissue low-temperature transport media		STM
Surgical excision in vacuum container		VAC
Swab		SWB
Other		ZZZ
Warm ischemia time
<2 min (including biopsy and direct resection)		A
2–10 min		B
10–20 min		C
20–30 min		D
30–60 min		E
>60 min		F
Unknown		X
Not applicable		N
Cold ischemia time
RT <2 min		A
RT 2–10 min		B
RT 10–20 min		C
RT 20–30 min		D
RT 30–60 min		E
RT 60 min–3 h		F
RT 3 h–6 h		G
RT 6 h–12 h		H
RT >12 h		I
2°C–10°C <60 min		E4
2°C–10°C 60 min–3 h		F4
2°C–10°C 3–6 h		G4
2°C–10°C 6–12 h		H4
2°C–10°C >12 h		I4
Unknown		X
Not applicable (e.g., autopsy)		N
Other		Z
Fixation/stabilization type
Nonaldehyde with acetic acid		ACA
Aldehyde based		ALD
Allprotect^® tissue reagent		ALL
Alcohol based		ETH
Nonbuffered formalin		FOR
Heat stabilization		HST
Snap freezing		SNP
Nonaldehyde based without acetic acid		NAA
Neutral buffered formalin		NBF
Optimum cutting temperature medium		OCT
PAXgene tissue		PXT
RNA Later		RNL
Vacuum technology stabilization		VAC
Unknown		XXX
Other		ZZZ
Fixation time
<15 min		A
15 min–1 h		B
1–4 h		C
4–8 h		D
8–28 h		E
28–52 h		F
52–76 h		G
72–100 h		H
>100 h		I
Not applicable		N
Unknown		X
Other		Z
Long-term storage
PP tube 0.5–2 mL	(−85) to (−60)°C	A
PP tube 0.5–2 mL	(−35) to (−18)°C	B
PP tube 0.5–2 mL	<−135°Ca	V
Cryotube^b 1–2 mL	LN	C
Cryotube^b 1–2 mL	(−85) to (−60)°C	D
Cryotube^b 1–2 mL	Programmable freezing to <−135°C	E
Plastic cryo straw	LN	F
Straw	(−85) to (−60)°C	G
Straw	(−35) to (−18)°C	H
Straw	Programmable freezing to <−135°C	I
PP tube ≥5 mL	(−85) to (−60)°C	J
PP tube ≥5 mL	(−35) to (−18)°C	K
Microplate well	(−85) to (−60)°C	L
Microplate well	(−35) to (−18)°C	M
Cryotube^b 1–2 mL	LN after temporary (−85) to (−60)°C	N
Straw	LN after temporary (−85) to (−60)°C	O
Paraffin block	RT or 2°C to 10°C	P
Paraffin block	(−35) to (−18)°C	U
Bag	LN	Q
Dry technology medium	RT	R
PP tube 40–500 μL	(−85) to (−60)°C	S
PP tube 40–500 μL	(−35) to (−18)°C	T
PP tube 40–500 μL	<−135°Ca	W
Original primary container	(−35) to (−18)°C or (−85) to (−60)°C	Y
Unknown		X
Other		Z

Codes in bold come from the LDMS. Volumes refer to container size.

^aTemperature <−135°C may correspond to LN vapor phase or to −150°C electrical freezer.

^bCryotube is defined as a tube that can be stored in LN either vapor or liquid phase.

Footnotes

Authors’ Contributions

L.W. and F.B.: Conceptualization. L.W.: Data curation and writing—original draft preparation. F.B., L.W., R.C., A.D.-W., W.G., M.K., L.L., W.M., I.M., R.A.-Q., K.S., P.S., S.S., D.S., J.T., R.T., and K.V.-N.: Reviewing and editing.

Author Disclosure Statement

There are no disclosures.

Funding Information

No funding was received for this article.

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