Gel Blood Collection Tube Affecting Test Results

American Journal of Pharmacy and Pharmacology
2016; 3(4): 40-45
http://www.aascit.org/journal/ajpp
ISSN: 2375-3900
Gel Blood Collection Tube Affecting
Test Results
Sawsan Mohammed Ali Hadi
Nutrition Research Institute, Baghdad, Iraq
Email address
[email protected]
Citation
Sawsan Mohammed Ali Hadi. Gel Blood Collection Tube Affecting Test Results. American
Journal of Pharmacy and Pharmacology. Vol. 3, No. 4, 2016, pp. 40-45.
Abstract
Keywords
Blood Collection Tubes,
Serum Separator Tubes (SST),
Gel Tube,
Vitamin A,
Vitamin D
Received: January 24, 2016
Accepted: May 27, 2016
Published: July 19, 2016
Accurate laboratory testing requires an understanding of the interactions between
collection tubes and blood specimens which can adversely affect the accuracy of
laboratory test results. The blood collection tubes like plain tube (containing no gel) used
for collection of serum for selected chemistry tests, and serum gel tube contains a clot
activator and gel separator used for various laboratory tests. Gel blood collection tube is
influenced by a number of variables, some of which are controlled by the tube
manufacturer, tube material, centrifugation speed and temperature; separator gel may
release materials that interfere with analytical assays. This study is an investigation of
the effect of gel blood collection tubes on vitamin A retinol, 25-OH vitamin D assays by
HPLC method.
1. Introduction
Fig. 1. Gel tube & plain tube (no gel).
American Journal of Pharmacy and Pharmacology 2016; 3(4): 40-45
“Serum-separator tubes, also known as SST, gel tubes, are
widely used in the clinical laboratory for routine collection
blood, it contain silica particles as a clot activator and a
special gel (the gel is composed of inert components, which
are part of a polyester-based proprietary formulation) [1]”.
The gel forms a physical barrier between serum and blood
cells during centrifugation, supernatant serum aspirated
directly from the collection tube.
“Gel blood collection tube is influenced by a number of
variables, some of which are controlled by the tube
manufacturer, tube material, centrifugation speed and
temperature; separator gel may release materials that
interfere with analytical assays [2]”.
“Several previous studies from USA, Japan, China,
Canada, and Iraq have shown the effects of interfering of gel
on clinical assay such as: gel may interfere with assays and
affecting analyte concentrations stability of blood analytes
after storage in BD SST tubes for 12 mo, Clinically
significant changes occurred only in 1,25-dihydroxyvitamin
D and retinol-binding protein [3]”.
“Separator gel may add materials, adsorb blood
components, or interact with protein and cellular components
[4]”. “Separator gels lead to a decrease in serum drug
concentrations [5]”. “Separator gels may release materials
(e.g., gel pieces and silicone oil) into the specimens and
spuriously interfere with assays [6]”.
“The separator gel components (SST) were the source of
interference in the quantitation of serum testosterone levels
[7]”. “Blood for 25-OH D measurement is probably best
collected into plain tubes without anticoagulants or gels [8]”.
“Gels have adverse effects on some steroid assays, including
HPLC methods for 25-OH D [9]”. “Unusual chromatography
– especially for 25OHD2 – was noted for some patient
samples. An investigation prompted the hypothesis that gel
separator in the blood collection tubes may cause interference
[10]”.
“It was noteworthy that some unexpected peaks appeared
in blood specimens collected in the particular blood
collection tube [11]”. “Serum 25OHD in VACUETTE tubes
with gel and clot activator, as measured by the Siemens
system, produced significantly higher values than did
samples collected in VACUETTE tubes with no additives
[12]”.
“Laboratories and tube manufacturers should be aware of
the limitation of using any tubes containing gel-separator
[13]”. “Vitamin D3 determination by HPLC cannot be
carried out with all gel tubes [14]”.
“Further studies have showed that an anomalous result
from gel tubes for vitamin D [15]”. “A recent blood
collection problem in our setting Troponin, Vitamin A, and E
[16]”.
“Blood collection tubes that contain separator gel
interfered with the quantification of steroid molecules 17hydroxyprogesterone and aldosterone by introducing
extraneous molecules that interfered with LC-MS/MS
41
analysis [17]”. “The serum TT3 concentrations obtained with
the SST tubes showed a significant positive proportional
difference compared with glass and Vacuette tubes [18]”.
“Do not use serum separator tubes for therapeutic drug
monitoring or toxicological analysis. The plastic serum
separator material extracts lipophilic substances (most
drugs), resulting in a falsely low drug concentration result.
Instead, collect the specimen in a plain red-top tube
containing no anticoagulants or preservatives [19]”.
The aim of the present study is to investigate if gel tubes
cause chromatographic interference with vitamin A (retinol)
and vitamin D (25OHD3) analysis by HPLC method.
2. Methods
Blood samples were collected from three healthy
volunteers into two tube types, gel separator serum tubes, and
plain tubes (tubes with no gel).
Blood collected in a plain tube (no gel), allow blood to clot
at room temperature for 30 minutes then centrifuge for 10
minutes to separate serum from clot.
Gel separator tube: gently invert the tube several times
(eight times). Allow blood to clot at ambient, temperature for
20-30 minutes. Centrifuge for 10 minutes to separate serum
from clot, If frozen serum is required, pour off serum into
plastic screw-cap vial and freeze [20].
Vitamin A assay by HPLC
Assay the concentration of vitamin A, by using HPLC
system knauer with a smart line UV detector 2500, pump
1000, and manager 5000. C18 column [250×4.6 mm (I.d.); 5
µm bead size]. The chromatographic separation was
performed by a mixture of methanol, water (95:5 by volume)
at a flow rate of 2.5 mL/min; Detection was monitored at 287
nm. The quantitative results were expressed as ug/dl vs.
control and calibrator.
Vitamin D assay by HPLC
Assay the concentration of vitamin D, RP- C18 column
(100 x 4.6 mm I.D.; particle size, 5 micron) at a flow-rate of
1 ml/min, the mobile phase was methanol. The eluate was
monitored with photodiode-array detector with wavelengths
265 nm. The quantitative results were expressed as ng/dl vs.
control and calibrator.
3. Results
Patient samples collected in gel separator tubes showed
that;
The chromatogram of retinol and 25OHD were Unusual,
they were not similar in shape, not identical the peaks differ
in height and there was a new peak present in gel tube sample
than the same patient samples collected in plain tubes (no
gel) (Fig. 2 and Fig. 3) and (Fig. 4 and Fig. 5)
The chromatography from the gel separator tubes may
affect the quantitation of both retinol and 25OHD.
42
Sawsan Mohammed Ali Hadi: Gel Blood Collection Tube Affecting Test Results
Fig. 2. Chromatogram of patient, serum collected into plain tube, vitamin A had a retention time of 2.3 minutes and 3.0 minutes for the internal standard,
retinol concentration ug/dl.
Fig. 3. Chromatogram of the same patient, serum collected into (gel tube), retinol Concentration ug/dl.
American Journal of Pharmacy and Pharmacology 2016; 3(4): 40-45
43
A new and unusual peak present between retinol and internal standard and the peaks show differ in height which effect the
concentration and vitamin result.
Comparing the chromatogram of vitamin D in serum collected in plain tubes and those collected in gel tubes, the results
were obtained as in (Fig3 and Fig4).
Fig. 4. Chromatogram of patient, serum collected into plain tube, vitamin D had a retention time of 3.7 minutes and 6.3 minutes for the internal standard,
vitamin D concentration ng/ml.
Fig. 5. Chromatogram of the same patient, serum collected into (gel tube), vitamin D3 Concentration ng/ml.
44
Sawsan Mohammed Ali Hadi: Gel Blood Collection Tube Affecting Test Results
A new and unusual peak present and the peaks show
differ in heights which effect the concentration and vitamin
result.
Tube comparison study for patient serum collected into
plain tube (no gel) and gel blood collection tubes show the
effects of gel which caused chromatographic interference
with retinol and 25OHD and detect the errors which affect
the ability of clinical laboratories to produce accurate
results.
4. Discussion
Ideally, the gel in blood collection tube should be inert to
the specimens collected in.
The gel components in blood collection tube may release
materials (gel pieces and silicone oil) into the specimens and
cause chromatographic interference with vitamin A (retinol)
and vitamin D (25OHD) by the HPLC method compared to
plain blood collection tube (no gel).
“Using gel vials for blood collection might cause disturbed
chromatograms caused by ingredients of the gel. We
recommend using EDTA plasma or serum vials without gel
[21]”.
Finally, evaluation of blood collection tubes by reference
clinical laboratories should be done to help in detecting tuberelated errors and interferences in test results which can
adversely influence patient outcomes and decrease laboratory
efficiency.
Table1. shows blood collection tubes-majority of
laboratories collect blood either in plain tubes or gel tubes.
For 25-OH vitamin D analysis;
Many laboratories collect blood specimen in a plain tube
(containing no gel) such as UWHC Lab, Regional Medical
Laboratory, NMS Labs, and Quest Diagnostics.
Other laboratories use gel blood collection tube and plain
tube (no gel) such as Mayo medical laboratories, Labcorp,
and Laboratory Corporation of America.
Other laboratories reject using gel blood collection tube
such as UWHC Lab.
For vitamin A (Retinol) analysis;
Many laboratories collect the blood specimen in a plain
tube (containing no gel) such as MLabs, Clinical Test
Catalog, Department of Pathology, and Lenco Clinical
Reference Laboratory
Other laboratories use gel blood collection tube and plain
tube (no gel) such as Mayo medical laboratories, UWHC
Lab, Labcorp, and Laboratory Corporation of America.
Other laboratories reject using gel blood collection tube
such as NMS Labs, MLabs, Clinical Test Catalog, and
Department of Pathology.
Laboratories should pay attention to the specimen types;
using wrong tube may make the blood sample unusable and
affect the clinical decision. Pre-analytical steps required for
laboratory testing results and any blood collection tubes
should be evaluated and compared the results obtained for
new and current tubes.
Table 1. Blood collection tubes specimen required in medical laboratories.
Medical laboratories
Mayo medical laboratories
http://www.mayomedicallaboratories.com/test-catalog/Specimen/60298 A
http://www.mayomedicallaboratories.com/test-catalog/Specimen/83670 D
UWHC Lab
http://www.uwhealth.org/lab-test-directory/ A
http://www.uwhealth.org/lab-test-directory/D
Regional Medical Laboratory
http://www.rmlonline.com/site/labtests/3603650?section_id=11 A
http://www.rmlonline.com/site/labtests/3603730 D
NMS Labs
http://www.nmslabs.com/uploads/PDF/2010_04_12_DBUpdate.pdf A
http://www.nmslabs.com/tests/Vitamin-D--25-Hydroxy--D2-and-D3---SerumPlasma/7628SP D
MLabs, Clinical Test Catalog, Department of Pathology
https://www.pathology.med.umich.edu/handbook/#/details/1020
Labcorp, Laboratory Corporation of America
https://www.labcorp.com/wps/portal/provider/testmenu
Lenco Clinical Reference Laboratory
http://www.lencolab.com/about_us/about_us.html
http://www.lencolab.com/doctors/test%20menue.html
Quest Diagnostics
http://www.questdiagnostics.com/home/physicians/testing-services/by-testname/vitamin D
Specimen Container;
Plain tube, red top tube, (No-Gel)
Gel blood collection tube, gel separation tube, serum gel, SST
Collection Container/Tube
Vitamin A (Retinol)
Collection container/Tube
25-OH vitamin D
Preferred: Plain tube
Acceptable: Gel separation tube
Preferred: Gel separation tube
Acceptable: Plain tube
Preferred; Plain tube
Acceptable; Gel separation tube
Acceptable; Plain tube
Rejection; SST
Preferred; Gel separation tube
Preferred; Plain tube
Rejection; Gel separation tube
Preferred; Plain tube
Preferred; Plain tube
Rejection: Gel separation tube
Preferred; Plain tube or Gel
separation tube
Preferred; Plain tube or Gel
separation tube
Preferred; Plain tube
Preferred; Plain tube
American Journal of Pharmacy and Pharmacology 2016; 3(4): 40-45
5. Conclusions
Gel collection tubes caused chromatographic interference
with vitamin A (Retinol) and vitamin D (25OHD)
quantitation by the HPLC method compared to serum
collection tubes by plain tube (containing no gel).
Gel blood collection tubes may adversely influence patient
outcomes, patient treatment, patient monitor, patient
diagnosis and decrease laboratory efficiency, and affect the
clinical decision.
Standardization of collection tube procedure will be an
important element in accurate analysis.
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