Stability of VACUETTE Lithium Heparin Separator tubes with modified centrifugation conditions
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1 Stability of VACUETTE Lithium Heparin Separator tubes with modified centrifugation conditions Background: Greiner-Bio-One, Austria has been selling plastic evacuated tubes (VACUETTE ) for venous blood collection since 19. The anticoagulant heparin activates antithrombins, thus blocking the coagulation cascade and producing a whole blood/plasma sample making it ideal for rapid analysis and analysis of blood from patients on anticoagulant therapy. VACUETTE Lithium Heparin Separator tubes contain a barrier gel in the tube. The specific gravity of this material lies between the blood cells and plasma. During centrifugation the gel moves upwards providing a stable barrier separating the plasma from cells. Plasma may be aspirated directly from the collection tube, eliminating the need for manual transfer to another container for analysis. [1] Study Objective: In order to shorten turn-around-time (TAT), laboratory professionals very often centrifuge a variety of blood collection tubes under the same centrifuge settings. This study was carried out to compare two different centrifugation conditions including stability of analytes for hours in order to improve laboratory efficiency by standardizing centrifugation conditions. [,3] Study design and procedure: Venous blood was collected from presumably healthy donors into four VACUETTE Lithium Heparin Separator tubes (7, 13/1, ml) using VACUETTE Safety Blood Collection Sets with Holder (item # ). Two different VACUETTE gel tubes (Sample A US P-Gel and Sample B European Gel) were included in this test. A discard tube was used to guarantee appropriate filling of the tubes. The order of collection was randomized. All tubes were inverted times immediately after blood collection. Tubes were then centrifuged with a Rotanta R centrifuge from Hettich at two different centrifugation conditions (see Table 1). Table 1: Centrifugation Sample RCF (x g) Time Temp [ C] Sample A g 1 min C P-Gel Sample A 1g 1 min C P-Gel Sample B g 1 min C Gel Sample B Gel 1g 1 min C All sample tubes were analyzed for the components listed below at the initial time point within hours of blood collection and after hours on the Siemens Advia analyzer and Abbott Architect analyzer using accompanying reagents from the instrument manufacturers. Between measurements, the samples were stored in the refrigerator at - C. The following parameters were measured: Alanine Aminotransferase (ALT) Alkaline Phosphatase (ALP) Aspartate Aminotransferase (AST) Beta-HCG Blood Urea Nitrogen (BUN) Calcium Chloride Cholesterol Cholinesterase (CHE) Creatinine page 1/1
2 Gamma Glutamyltransferase (GGT) Glucose Iron Lactate Dehydrogenase (LDH) Magnesium Phosphate Potassium Sodium Total Bilirubin Total Protein Triglyceride Troponin I Thyroid stimulating hormone (TSH) Urea Uric acid Vitamin B 1 Free Triiodthyronin (ft 3 ) Free Thyroxin (ft ) Results: Comparison analysis was performed at the initial time point and at hours. Statistical evaluation was performed with the T-test (α =.) using StatSoft, Version 9. Clinical evaluation was based on the allowed recommendation by the German Medical Association (RIILBÄK). [] No clinically significant differences were observed for either gel type with the two centrifugation settings of g for 1 minutes and 1g for 1 minutes at the initial time point or after hours. However, Glucose values were affected by decomposition in cells after hours. The use of a glycolysis inhibitor to compensate for this biochemical process would have been beneficial. Outlier testing was performed and showed that donor sample 1 (Sample A) for Alkaline Phosphatase at g for 1 min was an outlier and was, therefore, excluded from statistical analysis. For Lactate Dehydrogenase, an analysis error occurred with donor sample 3 at the centrifugation setting of g for 1 minutes at hours (Sample B); donor sample 3 at 1g for 1 min (Sample B) and donor sample 3 at 1g for 1 min (Sample B) at the initial time point and hours, all of which were excluded from statistical analysis. Conclusion: In this trial, the stability of the biochemical analytes was demonstrated when using the centrifugation conditions as tested (g for 1 min at C as well as 1g for 1 min at C) for VACUETTE Lithium Heparin Separator tubes to optimize TAT in routine laboratories. These centrifugation conditions for VACUETTE Lithium Heparin Separator tubes showed comparable results for the biochemical parameters tested at the initial time point and at hours. Differences in results were not clinically significant. References: [1] Instructions for Use. Evacuated Blood Collection System. For in vitro Diagnostic Use. Rev. 13. [] O Keane M. P., Evaluation of three different specimen types (serum, plasma, lithium heparin and serum gel separator) for analysis of certain analytes: clinical significance of differences in results and efficiency in use. Clin. Chem. Labor. Med. (), - [3] Thomas L., Laboratory and Diagnosis. Indication and Evaluation of Laboratory Results for Medical Diagnosis. 7 th edition. TH-Books. [] Guideline from the medical association in Germany for quality assurance of laboratory tests. German Medical Journal. Vol. 1, Issue 7.. page /1
3 Alanine Aminotransferase (ALT, GPT) Reference range: - U/l Sample A h Alanine Aminotransferase h GPT _1_A GPT 1_1_A Alanine Aminotransferase h GPT _1_A_ GPT 1_1_A_ Sample B h Alanine Aminotransferase h GPT _1_B GPT 1_1_B Alanine Aminotransferase h GPT _1_B_ GPT 1_1_B_ Alkaline Phosphatase: Reference range: 3-1 U/l Sample A h 1 1 Alkaline Phosphatase h AP _1_ A AP 1_1_A 1 1 Alkaline Phosphatase h AP _1_ A_ AP 1_1_A_ U/l Sample B h 1 Alkaline Phosphatase h AP _1_ B AP 1_1_B 1 Alkaline Phosphatase h AP _1_ B_ AP 1_1_B_ 1 1 page 3/1
4 Aspartate Aminotransferase (AST, GOT) Reference range: -3 U/l Sample A h 7 Aspartate Aminotransferase h 7 Aspartate Aminotransferase h GOT _1_A GOT 1_1_A GOT _1_A_ GOT 1_1_A_ Sample B h 7 Aspartate Aminotransferase h GOT _1_B GOT 1_1_B 7 Aspartate Aminotransferase h GOT _1_B_ GOT 1_1_B_ Beta-HCG Reference range: -.1 mie/ml Sample A h, 1, Beta-HCG h HCG _1_A HCG 1_1_A, 1, Beta-HCG h HCG _1_A_ HCG 1_1_A_ 1, 1, 1, 1, 1, 1, [mle/ml] 1, [mle/ml] 1,,,,,,,,,,, Sample B h, 1, 1, Beta-HCG h HCG _1_B HCG 1_1_B, 1, 1, Beta-HCG h HCG _1_B_ HCG 1_1_B_ 1, 1, 1, 1, [mle/ml] 1, [mle/ml] 1,,,,,,,,,,, RO page /1
5 Blood Urea Nitrogen (BUN) Reference range: 7-3 mg/dl Sample A h Blood Urea Nitrogen (BUN) h BUN _1_A BUN 1_1_A Blood Urea Nitrogen (BUN) h BUN _1_A_ BUN 1_1_A_ mg/dl 1 1 Sample B h Blood Urea Nitrogen (BUN) h BUN _1_B BUN 1_1_B Blood Urea Nitrogen (BUN) h BUN _1_B_ BUN 1_1_B_ Calcium Reference range:.1-.7 mmol/l Sample A h 3,, Calcium h Ca _1_A Ca 1_1_A 3,, Calcium h Ca _1_A_ Ca 1_1_A_,, 1, mmol/l 1, 1, 1,,,,, Sample B h 3, Calcium h 3, Calcium h Ca _1_B Ca 1_1_B Ca _1_B_ Ca 1_1_B_,,,, 1, 1, 1, 1,,,,, RO page /1
6 Chloride Reference range: 9-1 mmol/l Sample A h Chloride h Chloride h 1 Cl _1_A Cl 1_1_A 1 Cl _1_A_ Cl 1_1_A_ 1 1 mmol/l Sample B h Chloride h Chloride h 1 Cl _1_B Cl 1_1_B 1 Cl _1_B_ Cl 1_1_B_ 1 1 Cholesterol Reference range: 1- mg/dl Sample A h 3 3 Cholesterol h Chol _1_A Chol 1_1_A 3 3 Cholesterol h Chol _1_A_ Chol 1_1_A_ 1 mg/dl Sample B h 3 3 Cholesterol h Chol _1_B Chol 1_1_B 3 3 Cholesterol h Chol _1_B_ Chol 1_1_B_ RO page /1
7 Cholesterinesterase Reference range:.-11. ku/l Sample A h 1 1 Cholinesterase h CHE _1_A CHE 1_1_A 1 1 Cholinesterase h CHE _1_A_ CHE 1_1_A_ 1 1 [ku/l] [ku/l] Sample B h 1 1 Cholinesterase h CHE _1_B CHE 1_1_B 1 1 Cholinesterase h CHE _1_B_ CHE 1_1_B_ 1 1 [ku/l] [ku/l] Creatinine Reference range male: mg/dl Sample A h 1, Creatinine h 1, Creatinine h 1, Crea _1_A Crea 1_1_A 1, Crea _1_A_ Crea 1_1_A_ 1, 1, 1, 1, 1,, mg/dl 1,,,,,,,,,, Sample B h 1, Creatinine h 1, Creatinine h 1, Crea _1_B Crea 1_1_B 1, Crea _1_B_ Crea 1_1_B_ 1, 1, 1, 1, 1,, 1,,,,,,,,,, RO page 7/1
8 Gamma Glutamyltransferase (GGT) Reference range: - U/l Sample A h 1 9 GGT h GGT _1_A GGT 1_1_A 1 9 GGT h GGT _1_A_ GGT 1_1_A_ 7 7 U/l Sample B h 1 9 GGT h GGT _1_B GGT 1_1_B 1 9 GGT h GGT _1_B_ GGT 1_1_B_ Glucose Reference range: -11 mg/dl Sample A h 1 1 Glucose h Gluc _1_A Gluc 1_1_A 1 1 Glucose h Gluc _1_A_ Gluc 1_1_A_ 1 1 Sample B h 1 Glucose h 1 Glucose h 1 Gluc _1_B Gluc 1_1_B 1 Gluc _1_B_ Gluc 1_1_B_ 1 1 RO page /1
9 Iron Reference range: µmol/l Sample A h 3 3 Iron h Fe _1_A Fe 1_1_A 3 3 Iron h Fe _1_A_ Fe 1_1_A_ [µmol/l] 1 µmol/l Sample B h 3 3 Iron h Fe _1_B Fe 1_1_B 3 3 Iron h Fe _1_B_ Fe 1_1_B_ [µmol/l] 1 [µmol/l] Lactate Dehydrogenase Reference range: - U/l Sample A h 3 LDH h LDH _1_A LDH 1_1_A 3 LDH h LDH _1_A_ LDH 1_1_A_ Sample B h 3 LDH h LDH _1_B LDH 1_1_B 3 3 LDH h LDH _1_B_ LDH 1_1_B_ RO page 9/1
10 Magnesium Reference range:.7-1. mmol/l Sample A h 1, Magnesium h 1, Magnesium h Mg _1_A Mg 1_1_A Mg _1_A_ Mg 1_1_A_ 1, 1,,,,,,,,,,, Sample B h 1, 1, Magnesium h Mg _1_B Mg 1_1_B 1, 1, Magnesium h Mg _1_B_ Mg 1_1_B_,,,,,,,,,, Phosphate Reference range: mmol/l Sample A h 1, 1, Phosphate h P _1_A P 1_1_A 1, 1, Phosphate h P _1_A_ P 1_1_A_ 1, 1, 1, 1,,,,,,,,,, Sample B h 1, 1, 1, 1, Phosphate h P _1_B P 1_1_B, 1, 1, 1, 1, Phosphate h P _1_B_ P 1_1_B_,,,,,,,,,, RO page 1/1
11 Potassium Reference range: mmol/l Sample A h Potassium h Potassium h K _1_A K 1_1_A K _1_A_ K 1_1_A_ Sample B h Potassium h K _1_B K 1_1_B Potassium h K _1_B_ K 1_1_B_ Sodium Reference range: 13-1 mmol/l Sample A h 1 1 Sodium h Na _1_A Na 1_1_A 1 1 Sodium h Na _1_A_ Na 1_1_A_ Sample B h 1 Sodium h 1 Sodium h 1 Na _1_B Na 1_1_B 1 Na _1_B_ Na 1_1_B_ RO page 11/1
12 Total Bilirubin Reference range:.1-1. mg/dl Sample A h, 1, Total Bilirubin h TBili _1_A TBili 1_1_A, 1, Total Bilirubin h TBili _1_A_ TBili 1_1_A_ 1, 1, 1, 1, 1, 1, 1, 1,,,,,,,,,,, Sample B h, 1, Total Bilirubin h TBili _1_B TBili 1_1_B, 1, Total Bilirubin h TBili _1_B_ TBili 1_1_B_ 1, 1, 1, 1, 1, 1, 1, 1,,,,,,,,,,, Total Protein Reference range: - g/l Sample A h 1 Total Protein h TP _1_A TP 1_1_A 1 Total Protein h TP _1_A_ TP 1_1_A_ [g/l] Sample B h 1 Total Protein h TP _1_B TP 1_1_B [g/l] [g/l] 1 Total Protein h TP _1_B_ TP 1_1_B_ [g/l] RO page 1/1
13 Triglyceride Reference range: -1 mg/dl Sample A h 3 3 Triglyceride h Tri _1_A Tri 1_1_A 3 3 Triglyceride h Tri _1_A_ Tri 1_1_A_ Sample B h 3 3 Triglyceride h Tri _1_B Tri 1_1_B 3 3 Triglyceride h Tri _1_B_ Tri 1_1_B_ Troponin I Reference range: -. ng/ml Sample A h,3, Troponin I h Trop _1_A Trop 1_1_A,3, Troponin I h Trop _1_A_ Trop 1_1_A_,, [ng/ml],1 [ng/ml],1,1,1,,,, Sample B h,3 Troponin I h,3 Troponin I h, Trop _1_B Trop 1_1_B, Trop _1_B_ Trop 1_1_B_,, [ng/ml],1 [ng/ml],1,1,1,,,, RO page 13/1
14 Thyroid stimulating hormone (TSH) Reference range:.7-3. µu/ml Sample A h 7 TSH h 7 TSH h TSH _1_A TSH 1_1_A TSH _1_A_ TSH 1_1_A_ [µu/ml] 3 [µu/ml] Sample B h 7 TSH h TSH _1_B TSH 1_1_B 7 TSH h TSH _1_B_ TSH 1_1_B_ [µu/ml] 3 [µu/ml] Urea Reference range: 1- mg/dl Sample A h Urea h U _1_A U 1_1_A Urea h U _1_A_ U 1_1_A_ Sample B h Urea h U _1_B U 1_1_B Urea h U _1_B_ U 1_1_B_ RO page 1/1
15 Uric acid Reference range male: mg/dl Sample A h 1 Uric Acid h 1 Uric Acid h UA _1_A UA 1_1_A UA _1_A_ UA 1_1_A_ Sample B h 1 Uric Acid h UA _1_B UA 1_1_B 1 Uric Acid h UA _1_B_ UA 1_1_B_ Vitamin B 1 Reference range male: 1- ng/l Reference range female: 1- ng/l Sample A h 9 Vitamin B1 h Vit B _1_A Vit B 1_1_A 9 Vitamin B1 h Vit B _1_A_ Vit B 1_1_A_ 7 7 [pg/ml] [pg/ml] Sample B h 9 Vitamin B1 h 9 Vitamin B1 h Vit B _1_B Vit B 1_1_B Vit B _1_B_ Vit B 1_1_B_ 7 7 [pg/ml] [pg/ml] RO page 1/1
16 Free Triiodthyronin (ft 3 ) Reference range: pg/ml Sample A h, 3, ft3 h ft3 _1_A ft3 1_1_A, 3, ft3 h ft3 _1_A_ ft3 1_1_A_ 3, 3,,, [pg/ml], [pg/ml], 1, 1, 1, 1,,,,, Sample B h, 3, ft3 h ft3 _1_B FT3 1_1_B, 3, ft 3 h ft3 _1_B_ FT3 1_1_B_ 3, 3,,, [pg/ml], [pg/ml], 1, 1, 1, 1,,,,, Free Thyroxin (ft ) Reference range:.7-. ng/dl Sample A h 1, 1, ft h ft _1_A ft 1_1_A 1, 1, ft h ft _1_A_ ft 1_1_A_ 1, 1, 1, 1, [ng/dl], [ng/dl],,,,,,,,, Sample B h 1, 1, ft h ft _1_B ft 1_1_B 1, 1, ft h ft _1_B_ ft 1_1_B_ 1, 1, 1, 1, [ng/dl], [ng/dl],,,,,,,,, RO page 1/1
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