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Background Separation gels are often used in collection tubes, but adsorption of drugs onto the gel may cause falsely low concentrations in therapeutic drug monitoring. In this study, the stability of apixaban, edoxaban, rivaroxaban, flecainide, amiodarone, and desethylamiodarone was assessed in tubes, with and without gel separators. Methods Drug-free blood was spiked and stored for up to 7 d in non-gel tubes and gel tubes from two manufacturers (Vacuette and Vacutainer). The samples were analyzed in triplicates using ultra high pressure liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Results At ambient temperature conditions, the serum concentrations of apixaban, edoxaban, and rivaroxaban in a tube with acrylic-based gel had already decreased at baseline, while it took 6 h to observe the same result in a tube with olefin-based gel. At 4 °C, the reduction in serum concentration was considerably slower. For flecainide, the gel tube concentrations were stable at ambient temperature for 3 d, but decreased after 7 d in acrylic-based gel tubes. Amiodarone and desethylamiodarone stored in gel tubes at 4 °C showed decrease in concentrations after 24 h and 6 h, respectively. Conclusion Acrylic-based gel tubes should not be used for any of the tested drugs. Although olefin-based gel tubes may be used for anticoagulants and flecainide, it is advisable to prefer non-gel tubes as a general precaution.

Objectives Choosing the right device for blood collection is an issue that is frequently encountered in laboratory medicine. The purpose of this study was to compare the quality of the samples collected in terms of hemolysis index, pain intensity and tube filling rates of the samples drawn with BD Vacutainer® EclipseNeedle and BD Vacutainer Ultra-Touch™ Butterfly Blood Collection Set, which is currently used in laboratories. Methods Blood samples were drawn from the 44 apparently healthy adult individuals from different arm sites by routine phlebotomy into Citrate, SST and EDTA tubes in Karapınar State Hospital. K, P, AST, LDH tests and hemolysis index were analyzed by Mindray BS-800 chemistry analyzer. Complete blood count (CBC) was determined by Mindray BC 6000. Results There was no significant difference in K, P, AST, LDH and CBC values between the devices for hemolysis (p>0.05). No significant difference was found out with respect to visual hemolysis and hemolysis index, tube filling speed and pain intensity between the devices. Conclusions Our findings demonstrate that two sets may used for venipuncture blood collection without creating additional hemolysis risk.

Dual-column headspace gas chromatographic analysis with two flame-ionization detectors is a commonly used analytical technique for forensic blood ethanol quantitation. This technique is also applicable to the identification and quantitation of other volatile organic compounds such as methanol in biological samples. Compound identification by retention time is limited to those compounds with known retention times programmed into the instrument method. Historically, an early-eluting peak from an unidentified compound has been observed in both chromatograms from antemortem blood samples analyzed for ethanol concentration with this technique. The unidentified compound's retention time matches that of methanol on one column but not on the second column. This previously unidentified compound has been identified as isobutylene. The proposed source of the isobutylene contamination historically observed in antemortem blood samples collected in 10-ml gray-top blood collection tubes is the conventional rubber stopper. Isobutylene was detected in deionized water stored in each of the seven lots of 10-ml blood tubes tested; the expiration dates of the tubes tested spanned the years 2002-2022. Misidentification of isobutylene as methanol is possible when using a single-column gas chromatographic system. The presence of isobutylene in blood collected in a gray-top collection tube does not represent laboratory contamination, is not an interferent with blood ethanol quantitation, and does not affect the ethanol concentration in the blood. A 0.150 g/dl aqueous ethanol standard was stored in a gray-top tube to evaluate the potential impact of isobutylene on ethanol quantitation. The solution's average ethanol concentration measured after storage was 0.150 g/dl.

Background Direct membrane feeding assays (DMFA) are an important tool to study parasite transmission to mosquitoes. Mosquito feeding rates in these artificial systems require optimization, as there are a number of factors that potentially influence the feeding rates and there are no standardized methods that apply to all anopheline species. Methods A range of parameters prior to and during direct membrane feeding (DMF) were evaluated for their impact on Anopheles farauti sensu stricto feeding rates, including the starving conditions and duration of starving prior to feeding, membrane type, DMF exposure time, mosquito age, feeding in the light versus the dark, blood volume, mosquito density and temperature of water bath. Results The average successful DMFA feeding rate for An. farauti s.s. colony mosquitoes increased from 50 to 85% when assay parameters were varied. Overnight starvation and Baudruche membrane yielded the highest feeding rates but rates were also affected by blood volume in the feeder and the mosquito density in the feeding cups. Availability of water during the pre-feed starvation period did not significantly impact feeding rates, nor did the exposure duration to blood in membrane feeders, the age of mosquitoes (3, 5 and 7 days post-emergence), feeding in the light versus the dark, or the temperature (34 °C, 38 °C, 42 °C and 46 °C) of the water bath. Conclusion Optimal feeding conditions in An. farauti s.s. DMFA were to offer 50 female mosquitoes in a cup (with a total surface area of ~ 340 cm2 with 1 mosquito/6.8 cm2) that were starved overnight 350–500 µL of blood (collected in heparin-coated Vacutainer tubes) per feeder in feeders with a surface area ~ 5 cm2 (with a maximum capacity of 1.5 mL of blood) via a Baudruche membrane, for at least 10–20 min. Graphical Abstract

Background: Laboratory examination has several factors that can affect the results of the examination, one of which is the pre-analytic factor that can affect the results of erythrocyte examination is the ratio between blood volume and anticoagulant. If the blood volume is insufficient, the anticoagulant causes red blood cells to become krenated, and if the excess blood volume can cause anticoagulants it can cause blood clots. Research Objective: This study aims to determine the ratio of the number of erythrocytes in the blood sample volume of 3 mL, 2 mL, and 1 mL with anticoagulant K2EDTA. Research Methods: This study used primary data with a hematological examination at the UTD RSUD Dr. H. Abdul Moeloek Bandar Lampung. This type of research is quantitative using an observational analytic design with a crossapproach sectional through a hematological examination using the Hematology Alayzer Mindray BC-3600 with a sample size of 40 respondents who meet the inclusion and exclusion criteria. Results: The results of the mean examination of the number of erythrocytes between the blood volume of 1 mL, 2 mL, 3 mL with the anticoagulant K2EDTA had different results, at a volume of 3 mL showed the lowest results. Conclusion: There is no significant difference between the examination of the number of erythrocytes with the blood sample volume of 1 mL, 2 mL, and 3 mL in thetube vacutainer K2EDTA Keywords: Hematology Examinatio;, Blood Volume; K2EDTA.

Phosphatidylethanol (PEth) is a group of phospholipids formed in cell membranes following alcohol consumption by action of the enzyme phospholipase D (PLD). PEth measurement in whole blood samples is established as a specific alcohol biomarker with clinical and forensic applications. However, in blood specimens containing ethanol, formation of PEth may continue after sampling leading to falsely elevated concentrations. This study evaluated the use of dried blood spot (DBS) and microsampling specimens to avoid post-sampling formation of PEth. Filter paper cards and three commercial devices for volumetric microsampling of finger-pricked blood were assessed, using PEth-negative and PEth-positive whole blood fortified with 2 g/L ethanol. PEth (16:0/18:1) was measured by LC–MS/MS. Post-sampling formation of PEth occurred in wet blood and in the volumetric devices, but not filter paper cards, when stored at room temperature for 48 h. Addition of an inhibitor of PLD, sodium metavanadate (NaVO3), eliminated post-sampling formation during storage and drying. In conclusion, the present study confirmed previous observations that PEth can be formed in blood samples after collection, if the specimen contains ethanol. The results further demonstrated that post-sampling formation of PEth from ethanol also occurred with commercial devices for volumetric dried blood microsampling. In order for a PEth result not to be questioned, it is recommended to use a PLD inhibitor, whether venous blood is collected in a vacutainer tube or finger-pricked blood is obtained using devices for dried blood microsampling. Graphical abstract

Age and body weight at puberty were studied in two groups of Mutun helfen aged 4 to 7 months and comprising 13 heifers in each group. Group 1 heifers received nutritional supplementation et lating of a mixture of dry brewer's spent porin and palm kernel cake in a ratio of 1:1 addition to normal grazing. Group 1 theifers were grazed without any supplementation. From the age of 10 months and tasting for a period of 6 months, blood samples were collected twice weekly from each heifer via the jugular vein into heparinized Vacutabner tubes. Samples were centrifuged within 30 minutes of collection and the plasma stored frozen until assayed for progesterone using coat-2-count solid phase 125, radioimmunoassay kits. Body weights of all heifers were also recorded bi-weekly during the experimental period. Puberty was assumed to have occurred in any heifer when plasma progesterone values reached/exceeded i mg/ml. Results showed that Group 1 heifers were significantly younger (P< 0.01) at puberty than Group II heifers (12.25 + 0.20 months vs. 14.60+1.67 months) but their body weights did not differ significantly (90.50+ 1.82 kg & 85.20+ 0.87 kg for Groups I and II heifers, respectively). These results show that under improved feeding, puberty in Muturu heifers could be attained relatively at an early age. the first oestrus in the young female, is due to ovarian changes which are in turn controlled by complex endocrinological events. In the Mutury bull, Ezekwe (1992) observed that under improved husbandry and nutritional conditions, puberty was attained at an early age of 11 months and a body weight of 87 kg. The study also revealed that Muturu bulls could be used for breeding from the age of 15 months. Studies on female reproduction of indigenous Nigerian Cattle breed have concentrated largely on zebu (Bos indicus) cattle (Johnson and Gambo, 1979, Zakari et al., 1981; Oyedipe et al., 1982; Dawuda et al., 1989). There is thus a dearth of inforination on the reproductive characteristics such as puberty of indigenous Nigerian non-zebu cattle like the Muturu. The Muturu, which is the most widely distributed indigenous non-zebu cattle in the southeastern states of Nigeria, has a population of 42,000 out of an estimated total national population of 177,000. The Mutunu is highly tolerant to trypanosomiasis and is also well adapted to the hot humid conditions prevalent in the southern rainforest areas. Despite these obvious advantages, the Muturu is considered an endangered breed because of its rapidly declining numbers (Adeniyi, 1984). The need to document important aspects of its reproductive characteristics is therefore impelling. This study was thus undertaken to determine the age and body weight at which Muturu heifers attain puberty under improved husbandry and management conditions.

The Barricor tube (Becton Dickinson [BD], Sunnyvale, CA, USA) was recently developed to mechanically separate plasma by increasing the centrifugation rate. We compared the Barricor tube with existing serum- and plasma-based tubes based on 35 biochemical analytes and preanalytical turnaround time (TAT). Blood samples were collected from 30 healthy volunteers in a Barricor tube, serum separating tube (SST, Vacutainer SST II Tube 8.5 mL, #368972; BD), or plasma separating tube (PST, Vacutainer PST Tube 8.0 mL, #367964; BD) in random order. Next, 27 chemistry analytes, six immunochemistry analytes, and two cardiac markers were compared using Passing-Bablok regression and the Bland-Altman method. Preanalytical TAT was measured for each tube. The Barricor tube exhibited bias exceeding the desirable limit for nine and four analytes compared with the SST and PST, respectively. The Barricor tube lactate dehydrogenase value showed a bias of -10.29% and -9.86% compared with that of the SST and PST, respectively. The preanalytical TAT of Barricor tube was 8.8 minutes, which was the shortest among the three tubes. The clinical performance of the Barricor tube was equivalent to that of the SST and PST for most analytes, with an apparent advantage in preanalytical TAT. When using the Barricor tube, the reference range needs to be changed for some analytes that exceed the desirable bias limit.

Introduction: The blood sample is collected and transported to the laboratory for analysis, during this time the samples bares many extra-analytical factors prior to analysis. Contact time of plasma or serum with the blood cells is one of these factors because of ongoing cell metabolism as well as active and passive movement of analytes in cellular compartments. Aim and Objective: The aim of our study is to determine the stability of different seventeen biochemical parameters in primary collection (without separation of serum/plasma) and Secondary collection (with separation of serum/plasma) tube. Materials and Methods: Hundred patients’ venous blood sample were collected in plain vacutainer and fluoride vacutainer (primary tube), set of the seventeen biochemical analytes like sodium, potassium, ionized calcium, chloride, glucose, creatinine, urea, uric acid, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, total protein, albumin, bilirubin, triglyceride, total cholesterol and HDL-cholesterol tests were analysed. Once the tests were carried out from primary tube, approximately half serum was separated into two aliquots (secondary collection tube). All tubes were stored at 2 to 8° C. After 24 hr and 48 hr again same sate of tests were performed on the same equipment. Statistical analysis was done after that. Results: Out of seventeen analytes most analytes remained stable in both primary and secondary tubes up to 48 hr at 2 to 8° C, these includes chloride, creatinine, urea, uric acid, total bilirubin, total protein, albumin. alkaline phosphatase (ALP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT). Plasma glucose decreased in both primary and secondary collection tube but concentration is within significant change limit (SCL). While serum potassium is increase 28.84% at 24 hr and 75.58% at 48 hr; concentration is also out of SCL range in primary collection tube even though samples stored at 2 - 8⁰ C. Conclusion: Most biochemical analytes will remain stable for 48 hr at 2 - 8⁰ C, only few analytes like plasma glucose and serum potassium requires stringent control before serum-clot separation. Keywords: Collection tube, Pre-analytical factors, Specimen stability.

Background Diabetes mellitus (DM) patients show a significant derangement in various hematological parameters including changes affecting the red blood cells (RBCs). All these derangements have an imposing effect on any of the RBC indices. Thus, the main aim of this study was determining the RBCs parameters and their correlation with renal function, and also the magnitude of anemia in DM patients in comparison with healthy controls. Methods A comparative cross-sectional study was conducted at the chronic illness clinic of University of Gondar Hospital from January to April 2020. A total of 246 participants (164 DM and 82 controls) were selected using a simple random sampling technique. Data were collected using a pretested structured questionnaire. Five milliliters venous blood was collected by vacutainer blood collection technique. RBC parameters and renal function tests were determined by using Sysmex KX21N and BS-200E Mindray analyzers, respectively. The data were tested for normality using Shapiro-Wilk and Kolmogorov-Smirnov tests. Independent sample t-test and Pearson's and Spearman correlation statistics were used to analyze variables. A p-value ˂0.05 was considered as statistically significant. Results The mean hemoglobin (Hgb) concentration was 13.8±0.7 and 15±1.3 among DM and controls, respectively. RBC (p=0.014), Hgb (p ˂0.001), hematocrit (Hct) (p ˂0.001), and mean cell volume (MCV) (p ˂0.001) were significantly lower in DM patients as compared to controls. On the other hand, mean cell hemoglobin concentration (MCHC) was significantly increased in DM patients (p ˂0.001) than the controls. Besides, a significant negative correlation was found between Hgb and creatinine (Cr) in DM patients. Conclusion The mean values of RBC parameters (RBC, Hgb, Hct, and MCV) for DM patients were found significantly lower than the control groups. Besides a significant negative correlation was found between Cr and RBC indices (RBC, Hgb, Hct, and MCV) in DM patients. It is, therefore, suggested that RBC parameters abnormalities should be evaluated and treated periodically in DM patients for better prognosis and quality of life.

Background: Hepcidin is known to be the central regulator of iron homeostasis in the body. It is up-regulated by inflammation and downregulated by anemia. CKD is a state of chronic inflammation seen in kidney. Previous work has shown that serum hepcidin levels were increased in patients with CKD. This was surprising as these patients had a chronic inflammatory state and co-existent anemia. Aim and Objectives: The aim of the study is to estimate the levels of hepcidin in CKD patients and to check the correlation of hepcidin to inflammation in chronic kidney disease. Methods: This cross-sectional study was conducted at the Department of Biochemistry, Central Laboratory, Sree Balaji Medical College and Hospital, Chromepet, Chennai during January 2017 - June 2018 among 50 patients of chronic kidney disease in the age group of 18-60 years. The blood samples were collected using vacutainer system. Samples for serum hepcidin, ferritin and hsCRP were collected in red topped plain vacuum tube. The samples were centrifuged at 3000 rpm for 15 minutes. The samples were then processed, and values were obtained. The data were analysed using SPSS package. Results: The mean values of s. Hepcidin, s. ferritin and hsCRP levels were found to be increased in the study population. The mean value of s. hepcidin was found to have strong positive correlation with the mean values of s. ferritin and hsCRP with r-value > 0.7. Conclusion: Hepcidin levels are elevated in CKD and hepcidin is a predictor of inflammation since it correlated well with the inflammatory markers hsCRP and ferritin levels.

Essential and nonessential element concentrations in human blood provide important information on the nutritional status of individuals and can assist in the screening or diagnosis of certain disorders and their association with other causative factors. A simple and sensitive method, suitable for use with small sample volumes, for quantification of multiple trace element concentrations in whole blood and plasma has been developed using inductively coupled plasma-mass spectrometry. Method validation was performed using standard reference materials of whole blood and serum using varying sample treatments with nitric acid, water and hydrogen peroxide. The method was applied to quantify the trace element concentrations in whole blood and plasma samples (0.1 mL) from 50 adult blood donors in Queensland. The whole blood sample (5 mL) was collected in Vacutainer tubes with K2EDTA as anticoagulant. The developed method was able to quantify, in blood and plasma samples over a wide range of concentrations, several essential elements: cobalt, copper, zinc, iron, manganese and selenium; the nutritionally probably essential elements vanadium and strontium; and nonessential elements including lead, cadmium, arsenic, caesium, barium, thallium and uranium. Significant differences (P

Blood specimens are used to study the aetiology, diagnosis, treatment, prognosis, and prevention of blood diseases that includes research purposes. The primary health care worldwide calls for ideal systems that can provide accurate results but it is challenging to analyse the samples immediately. Thus, the primary objective of this study was to determine the changes in the red cell indices when stored at room temperatures. To accept such samples that have delayed after collections for some time as from zero (0) hours, at 6 hours, 12 hours, 24 hours and 48 hours requires testing and confirming the results validations. Therefore, this research analysed samples of blood to validate the results after storage at room temperature in different hours within 48 hours of storage at room temperature. Samples collected from donors aged 16-65 years at Kisii Teaching and Referral Hospital in Kisii County, Western Kenya were tested in this study. The technique of Flow cytometry was used to analyse results of blood obtained through Venepuncture procedure from donors and placed in 4 ml Ethylene Diamine Tetra Acetic (EDTA) Vacutainer tubes of standardised anticoagulant ratio. A cross-sectional study with laboratory experiment was employed to analyse the data between the month of April and August 2018. Percentile male and female blood samples were analysed at 48% and 52% respectively. The analytes of obtained were used to determine the two red blood cell parameters in controlled room temperature within 48 hours on Packed Cell Volume (PCV) and Total Red Cell counts. Statistical analysis with computerized applications package (SPSS, version 21.0) was used to determine the changes that occurred during the study with repeated Analysis of Variation (ANOVA). Comparative mean values were calculated for each blood samples. The results were insignificant at sixth and twelfth hours of storage for most indices except from the RBC total counts, which had a deviation between 6-48 hours. In conclusion blood sample analysis is safe for tests results up 12 hours of storage at room temperature when not tested immediately after collection. This study recommends further finds for Reticulocytes Production Index to assess bone marrow activities.

Background Some therapeutic drugs are unstable during sample storage in gel tubes. BD Vacutainer® Barricor™ Plasma Blood Collection Tube with nongel separator was compared with plasma gel tubes, BD Vacutainer PST™, PST II, and BD Vacutainer Serum Tube for acetaminophen, salicylate, digoxin, carbamazepine, phenytoin, valproic acid, and vancomycin during sample storage for up to 7 days. Methods Seven hospital sites enrolled 705 participants who were taking at least one selected drug. The study tubes were collected and tested at initial time (0 h), after 48 h of storage at room temperature and on day 7 (after additional 5 days of refrigerated storage). The performance of BD Barricor tube was evaluated for each drug by comparing BD Barricor samples with samples from the other tubes at 0 h from the same participant; stability was evaluated by comparing test results from the same tube at 0 h, 48 h, and 7 days. Results At 0 h, BD Barricor showed clinically equivalent results for selected therapeutic drugs compared with the other tubes, except phenytoin in BD PST. Phenytoin samples ≥20 µg/mL in BD PST had 10–12% lower values than samples in BD Barricor. During sample storage, all selected drugs remained stable for 7 days in BD Barricor and in serum aliquots. In BD PST, all drugs remained stable except phenytoin and carbamazepine and in BD PST II except for phenytoin. Conclusion The BD Barricor Tube is effective for the collection and storage of plasma blood samples for therapeutic drug monitoring without sample aliquoting.

Testing for Human Immunodeficiency Virus (HIV), widely distributed in sub-Saharan Africa since it is mainly invasive but, could be non-invasive and quick also, reducing waiting time especially when required for presurgical procedures. This study determined the HIV status of patients using Urine screening test method and to compare its performance to blood-based testing methods. The routine pre and post-test counselling for HIV screening were done for all provider-initiated HIV testing using blood-based screening methods. Due to the cost and unavailability of enough urine testing kits, only patients who tested positive for HIV with blood-based methods and were scheduled for surgery or a surgical procedure were enrolled in the study. Informed consent was obtained. Paired urine and blood samples were collected at the same visit into clean universal bottles and analyzed immediately. A colloidal gold enhanced rapid immuno-chromatographic assay (Alliance Biomedical) kit for the rapid qualitative detection of antibodies to Human Immunodeficiency Virus (HIV) I and II in urine were used in comparison to the standard HIV testing of ante-cubital venous blood collected in EDTA vacutainer and analyzed using Determine (T) HIV 1 and 2 in vitro qualitative immunoassay strip, UNI GOLD rapid test kit and the Chembio HIV 1/2 STAT PAK assay strip. A total of 7568 patients were tested for routine provider-initiated HIV testing, 521 tested HIV positive. There were 105 (20.15%) males and 416 (79.85%) females, age ranged from 15 years to >80 years. Most of the surgeries performed were Caesarian section 93 (37%), Hernia 55 (22%), Lumps 48 (19%), Acute appendicitis 33 (13%), Uterine fibroids 10 (4%), Ruptured ectopic pregnancy 2 (1%) and others (Intestinal obstruction, Postoperative adhesions, Ingrown toe nails, Breast abscess, Hemorrhoids, Anal fissures etc.) 10 (4%). DETERMINE RAPID HIV TEST METHOD USING BLOOD: A total of 521 HIV positive samples were tested, 502 (96.35%) tested HIV positive and 19 (3.65%) tested HIV negative. These 19 HIV negatives were re-tested with Stak Pak: 19 (100%) tested HIV positive. UNI GOLD HIV TEST METHOD USING BLOOD: A total of 521 HIV positive samples were tested, 521 (100%) tested HIV positive. URINE TESTING METHOD: A total of 251 (48.18%) of the 521 HIV positive patients were scheduled to undergo a surgical procedure. These were re-tested using the Urine testing method, 235 (93.63%) tested HIV positive while 16 (6.37%) tested negative. The blood sample of the 16 who tested negative using the Urine testing method was subjected to confirmatory test using Stat Pak and all 16 (100%) tested HIV positive. The specificity for Unigold and Determine blood testing was 100%. All three tests had a Positive Predictive Value (PPV) of 100% while the Negative Predictive Values (NPV) were 100% and 99.73% for Unigold and Determine respectively. The use of Urine HIV testing method compared well to the blood HIV testing methods and could be a better non-invasive sample method for screening of HIV/AIDS in the population especially among surgeons’ pre-surgical procedures.

Introduction: Sample for plasma glucose estimation is taken in fluoride Vacutainer and that is to prevent glycolysis and so that accurate glucose estimation can be done. Objectives: The objective of the study is to study various glycolytic inhibitors for preservation of blood glucose and other common clinical biochemical parameters. Materials and Methods: This prospective study was conducted at new civil hospital Surat. Study includes total 100 participants. We have prepared 5 mmol/L DL Glyceraldehydes containing Vacutainer. Blood sample from all participants were taken in 3 Vacutainer like plain Vacutainer, fluoride Vacutainer and DL Glyceraldehydes containing Vacutainer. Various biochemical investigations were performed from all above collected blood samples to see the difference of significant among them by calculating p value. P value less than 0.05 was considered as a difference of significant. Results: There is no significant changes in results of glucose between fluoride and glycrealdehyde containing Vacutainer. Biochemical parameter like SGPT, Creatinine, Total Bilirubin, Albumin, cholesterol, Total protein, Uric acid, electrolytes also does not show any significant difference between DL-Glyceraldehyde and plain Vacutainer. The Result of Serum Creatinine was found to be high from Glyceraldehyde containing Vacutainer as compared to plain Vacutainer and difference among them is highly significant.(p Conclusion: From our study we would like to conclude that. DL -Glyceraldehyde containing vial for many biochemistry related parameter analysis is better option as it is save additional use of vaccutte, except for serum creatinine (by alkaline picrate method). So DL Glycrealdehyde cannot be solely used as alternative for plain and fluoride containing Vacutainer. Keywords: DL-Glyceraldehyde, Glycolysis, Fluoride, Creatinine, Vacutainer.

Adenosine is a purine ribonucleoside with important roles in various physiological processes. A number of studies have indicated the importance of adenosine in cardiovascular diseases including syncope, however, the accurate determination of adenosine in human blood is challenging due to the molecule’s instability. In the present study, we report a simple method for the pre-treatment of blood samples and the development of a fast and efficient hydrophilic interaction chromatographic tandem mass spectrometry method for the analysis of adenosine in patient blood. During collection, samples were mixed directly with a solvent mixture containing 95% acetonitrile and 10 mM ammonium formate in a Vacutainer tube, resulting in successful prevention of adenosine metabolic processes and direct blood sample deproteinization. The method was validated according to bioanalytical industry guidelines and found to be accurate, repeatable, specific and sensitive with LLOQ 0.005 μg/mL, thus allowing its application in the analysis of real clinical samples.

Ceftiofur is an important broad-spectrum 3 rd generation cephalosporin antibiotic. Owing to its time-dependent antimicrobial actions, the length of time of being above bacterial MIC is the critical point in using ceftiofur for chemotherapy rather than its peak of concentration. Consequently, this experiment was carried out to evaluate, for the first time, the pharmacokinetics of the long-acting ceftiofur crystalline acid-free form (ceftiofur-CAF) in camels. Ceftiofur-CAF 200 mg/ml suspension sterile solution was injected i/m at a dose 6.6 mg/kg. Blood samples were collected from the jugular vein in vacutainer tubes at 0, 0.13, 0.25, 0.5, 1, 2, 4, 8, 12, 24, 48, 72, 96, 120, and 144 hours after administration of the drug. Ultrahigh Performance Liquid Chromatography-Mass Spectrometry (UPLC MS/MS) was used to measure serum concentration. Pharmacokinetic modeling was by a two-compartment model. Pharmacokinetics of ceftiofur-CAF after single i/m injection in she-camels was best modeled in the two-compartment model, where the drug slowly distributed to a second compartment with poor tissue penetration and high preference to the central compartment. In this study, the maximum plasma concentration (C max ) was 9.29±0.42 μg/ml at T max equals 9.41±1.35 h. The area under the curve (AUC 0-∞ ) was 354.1±57.22 μg/ml*h. The distribution and elimination half-lives were 7.42 and 46.13 h, respectively. The mean residence time (MRT) was 42.01 h. Compared with the rapidly absorbed form of ceftiofur (ceftufor-RAF) in camels, there was almost similar maximal serum concentration but with delayed time to maximal concentration (T max ), longer means residence time (MRT) and higher distribution and elimination half-lives. In terms of antibacterial efficacy, ceftiofur-CAF stayed above a previously recommended level of 0.2 μg/ml for 7 days, which can be achieved after a single i/m injection of 6.6 mg/kg. The obtained pharmacokinetics data in camels recommends repeated administration of 2 days apart for bacteria requiring MIC levels above 2 μg/ml. Key words: Ceftiofur; pharmacokinetics; camel; cephalosporins FARMAKOKINETIKA DOLGODELUJOCE CEFTIOFURNE KRISTALINICNE PROSTE KISLINE PRI SAMICAH ARABSKIH KAMEL ( Camelus dromedarius ) Izvlecek: Ceftiofur je pomemben sirokospektralni antibiotik 3. generacije cefalosporinov. Zaradi njegovih casovno odvisnih protimikrobnih ucinkov je cas, ko je raven ceftiofura nad bakterijskim MIC in ne pri njegovem vrhu koncentracije kriticna tocka pri uporabi tega antibiotika. Poskus je bil izveden z namenom ovrednotenja farmakokinetike dolgo delujoce ceftiofurjeve kristalinicne brezkislinske oblike (ceftiofur-CAF) pri kamelah. Ceftiofur-CAF v koncentraciji 200 mg/ml suspenzije sterilne raztopine smo injicirali i/m v odmerku 6,6 mg/kg. Vzorci krvi so bili zbrani iz vratne vene v vakuumskih epruvetah ob injiciranju antibiotika in nato 8, 15 in 30 minut po injiciranju ter 1, 2, 4, 8, 12, 24, 48, 72, 96, 120 in 144 ur po injiciranju antibiotika. Za merjenje serumske koncentracije je bila uporabljena tekocinska kromatografija ultra visoke locljivosti ( UPLC MS/MS ). Farmakokineticno modeliranje je bilo izvedeno z dvokomponentnim modelom. Farmakokinetiko ceftiofur-CAF-a po enkratnem i/m injiciranju v kamele je bilo najbolje modelirati v modelu z dvema predelkoma, kjer se je zdravilo pocasi razdeljevalo v drugi predelek s slabo penetracijo v tkiva in veliko prednostjo do osrednjega predelka. Najvisja koncentracija antibiotika v plazmi (C max ) je bila 9,29 ± 0,42 μg/ml pri T max 9,41 ± 1,35 ure. Povrsina pod krivuljo (AUC0-∞) je bila 354,1 ± 57,22 μg/ml*h. Razpolovni cas razporeditve in izlocanja je bil 7,42 oziroma 46,13 ure. Povprecni cas prisotnosti antibiotika (MRT) je bil 42,01 h. V primerjavi s hitro absorbirano obliko ceftiofurja (ceftufor-RAF) pri kamelah je bila skoraj podobna najvecja koncentracija v serumu, vendar z zakasnjenim casom do najvecje koncentracije (T max ), daljsim casom zadrževanja (MRT) in vecjim razpolovnim casom porazdelitve in izlocanja. Ceftiofur-CAF ostal dni nad predhodno priporoceno ravnijo ucinkovitosti 0,2 μg/ml kar 7 dni, kar je bilo mogoce doseci po enkratni i/m injekciji 6,6 mg/kg. Pridobljeni podatki o farmakokinetiki v kamelah priporocajo veckratno dajanje v razmaku 2 dni za bakterije, ki potrebujejo ravni MIC nad 2 μg/ml. Kljucne besede: Ceftiofur; farmakokinetika; kamela; cefalosporini

Drugs may potentially adsorb to blood collection tubes containing gel separators in the preanalytical phase of therapeutic drug monitoring. The aim of this study was to compare measured concentrations of 28 psychoactive drugs and 13 metabolites in spiked serum samples stored on standard (plain) tubes versus barrier gel tubes during a 2-6-day period at room temperature.Drug-free ("blank") serum samples spiked with mixes of antidepressants, antipsychotics, or mood stabilizers (valproic acid and lamotrigine), including relevant metabolites, were transferred to tubes with and without gel, that is, BD Vacutainer SST II Advance gel tubes and BD Vacutainer Glass Serum Tubes (Becton-Dickinson Company, Plymouth, United Kingdom). Mean serum concentrations of the drugs or metabolites measured by ultra-high performance liquid chromatography-tandem mass spectrometry analyses of protein-precipitated samples were compared after storage on plain or gel tubes at 3 time points (day 0, day 2/48 hours, and day 6/144 hours) in room temperature.Mean serum concentrations of all antidepressants, except for one metabolite, and 13 of 18 antipsychotic drugs were significantly lower in gel tubes compared with plain tubes after 2 days of storage (2%-28% lower, P0.05). After 6 days of storage, mean serum concentrations of all antipsychotic drugs and antidepressants were significantly lower in gel tubes versus plain tubes (9%-49% lower, P0.02), except for amisulpride and O-desmethylvenlafaxine. Serum concentrations of the mood stabilizers were not significantly different in gel tubes compared with plain tubes (P0.1). There was a clear relationship between log P (partition coefficient) and residual serum concentrations during gel tube storage (r -0.50 and -0.42 at day 2 and day 6, respectively; P0.02).This study shows that storage on gel for more than 2 days significantly decreases the serum concentrations of antidepressant and antipsychotic drugs as compared to storage in plain tubes. Thus, using tubes with gel separators in the therapeutic drug monitoring of psychoactive drugs should be reconsidered.

Background The target values for plasma glucose concentrations for the investigation and diagnosis of diabetes and impaired fasting glucose, and the realization that small incremental changes in glucose concentration increase the risk of adverse events, has led to greater focus on laboratory glucose results. Although analytical methods show acceptable precision, the control of preanalytical error due to the stability of glucose remains problematic. The aim of this study was to compare glucose concentrations in 3 different and commercially available blood tubes, with analysis and storage under current practices and conditions. Methods Blood samples for glucose were obtained from consenting patients attending the Diabetic Clinic at the Royal Victoria Hospital, Belfast. Blood was collected into BD Vacutainer® Barricor™ Lithium Heparin tubes, BD Vacutainer Fluoride EDTA tubes, and Greiner Vacuette® FC-Mix (sodium fluoride/citrate/Na2EDTA) tubes in that order. The Barricor tubes were immediately centrifuged at 4000g for 3 min. All samples were then sent to the Biochemistry Laboratory for analysis on the same day, and again the following day after storage at 4 °C. Results There was no significant difference in mean glucose concentrations between immediately centrifuged Barricor and FC-Mix tubes when analyzed on day 0. Both tube types demonstrated higher mean glucose concentrations than traditional fluoride EDTA (F/EDTA) samples. Conclusions Both immediately separated Barricor and citrated FC-Mix plasma preserve glucose concentrations to the same extent, and better than F/EDTA preservative. These newer technologies involved offer pragmatic solutions to improved glucose analysis, allowing laboratories to choose the best option given the source of their samples.

Autologous conditioned serum (ACS) is commonly used for the treatment of human and equine osteoarthritis (OA). The objective of this study was to investigate the influence of incubation time and incubation tube on concentrations of interleukin-1 receptor antagonist (IL-1ra), interleukin-1 beta (IL-1β), and insulin-like growth factor-1 (IGF-1) in ACS of horses with and without OA. Blood samples of 7 control horses and 6 horses with naturally occurring OA were obtained. The concentration of serum IL-1ra, IL-1β, and IGF-1 was determined after 12, 24, and 36 hours of incubation using commercial serum vacutainer glass tubes (incubated serum [IS]) or a specific equine ACS system. There was no significant difference in IL-1ra, IL-1β, and IGF-1 concentrations between IS and ACS samples detectable at 24 or 36 hours of incubation. The ACS of horses with OA and control horses did not contain significantly different cytokine or growth factor concentrations. In conclusion, an incubation time exceeding 24 hours does not lead to a significant increase in cytokine and growth factor concentrations in equine ACS. Incubation of whole blood in commercial serum glass tubes has similar effects on the IL-1ra, IL-1β, and IGF-1 concentrations when compared with the incubation in specialized equine ACS kits. Interleukin-1 receptor antagonist, IL-1β, and IGF-1 concentrations in ACS of OA horses is not significantly increased when compared with control horses.

Aim: The effect of citrate phosphate dextrose adenine-1 on the hematological parameters of stored Nigerian indigenous dog's blood with plastic vacutainer was investigated. This was done in view of determining the viability and stability of the studied parameters for blood banking purpose. Till date, there is no literature on the stability of whole blood of Nigerian indigenous dogs for blood banking purposes. Materials and Methods: A total of six apparently adult healthy dogs were sampled, and their blood was stored at 4°C and analyzed for their packed cell volume (PCV), hemoglobin (Hgb) concentration, red blood cell (RBC) count, total and differential white blood cell (WBC) count, platelet count (PC), mean corpuscular values (mean corpuscular Hgb [MCH], MCH concentration, and mean corpuscular volume [MCV]), blood potency of hydrogen (pH), and erythrocyte sedimentation rate (ESR) over a period of 14 days. Results: Significant changes were observed in some of the studied parameters. Of the 14 days' study period, PCV, Hgb concentration, total WBC count, PC, and neutrophil count showed no significant (p≥0.05) difference until day 2 post-storage (PS), while RBC count, ESR, MCV, and lymphocyte count showed no significant (p≥0.05) variation up until day 3 PS. Blood pH showed no significant (p≥0.05) variation within 24-h PS but was significantly (p≤0.05) higher than obtained values on days 1, 3, 7, 10, and 14 PS. Conclusion: Based on our finding, we could suggest that hematological laboratories and hemotherapists could use canine blood stored in a plastic vacutainer for 2-3 days.

Context: Elevated blood pressure and alterations in lipid and lipoproteins play a major role in the development and progression of cardiovascular diseases. Aim: This study is aimed at determining the blood pressure and lipid profile in automechanics. Settings and Design: A total of 120 male subjects between 18 and 55 years of age comprised 60 automechanics and 60 age-matched occupationally unexposed control subjects in Emene, Enugu State, Nigeria. Subjects and Methods: Four milliliters of fasting blood samples was collected from all subjects, 2 mL of blood was dispensed into K2-EDTA vacutainer tube for blood lead analysis, while the other 2 mL was dispensed into plain vacutainer tube, allowed to retract, centrifuged, and the serum used for serum lipid profile analysis. Blood pressure was measured using aneroid sphygmomanometer. Statistical Analysis: Statistical Package for Social Sciences (SPSS) version 21.0 was used for data analysis. Results: Automechanics had significantly higher values of systolic and diastolic blood pressures compared with the controls. Serum levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), very low-density lipoprotein cholesterol (VLDL-C), and triglyceride (TG) were also significantly higher in the automechanics compared with the controls (P < 0.05); serum high-density lipoprotein cholesterol (HDL-C) level, however, did not differ significantly between the two groups (P > 0.05). Blood lead level showed a significant positive correlation (P < 0.05) with systolic and diastolic blood pressures, serum TC, LDL-C, VLDL-C, and TG, with no significant correlation with serum HDL-C (P > 0.05) in the automechanics and no correlation in the controls. Conclusion: The result of this study indicates that blood pressure is elevated and lipid profile altered in automechanics which suggests that these groups of workers are prone to increased risk of developing hypertension and cardiovascular disorders due to occupational exposure to lead.

Background The use of anticoagulants may influence the composition of blood cells and interfere with plasma levels of IL-1ra when unprocessed EDTA blood samples are stored for long periods of time. Methods Blood was drawn into EDTA and heparinized blood collection tubes from 11 HIV-1 negative men participating in the Multicenter AIDS Cohort Study (MACS) and 4 healthy volunteers. The blood was processed according to the experiments detailed in the method and after incubation; supernatants were collected and stored at -70 °C until batch testing using IL-1ra ELISA. Results There was no difference between the levels of IL-1ra in EDTA blood collected into plastic and glass tubes (p = .911). There were significant increases from baseline levels of IL-1ra (p ≤ .05) after 24 h incubation for diluted whole blood and PBMC supernatants but not for granulocytes supernatants. Conclusion EDTA as an anticoagulant influences the blood concentrations of IL-1ra in unprocessed blood. Thus, EDTA blood is not a suitable specimen for measurement of IL-1ra. Other types of anticoagulated blood should be processed within one hour of draw whenever measuring plasma levels of IL-1ra.

AIM This study aims to test the feasibility and effectiveness of a novel pacifier-based saliva collection method on children and infants in comparison to an oral swab-based saliva collection method. MATERIALS AND METHODS This study was performed during spring 2018 in a clinical non-sponsored setting at Queen Silvia Children's Hospital pediatric emergency ward. Saliva collection was performed by comparing oral swab (Salimetrics® SalivaBio's Children's Swab) with a pacifier-based saliva collection method (Salivac®). All participating children used both saliva collection systems. The amount of saliva collected in 2 minutes was measured. The amount of time needed for the healthcare professional was recorded. Parental preference was evaluated by a questionnaire. RESULTS No statistically significant difference was observed in collected saliva (174 µL for pacifier-based saliva collection and 158 µL for oral swab). The healthcare professional spent significantly less (p < 0.001) mean time with the pacifier-based saliva collection method than with the oral swab (31 vs 150 sec). A total of 48 out of the 52 caretakers preferred the pacifier-based saliva collection method compared to the oral swab. CONCLUSION The novel pacifier-based saliva collection method proved to be a feasible, appreciated, and effective way of collecting saliva that simplifies the saliva collection method among children and infants. CLINICAL SIGNIFICANCE The pacifier-based saliva collection method simplifies saliva testing. The closed vacutainer system minimizes the risk of saliva contamination and opens up for novel home testing strategies.

Background Oncology patients have frequent venipunctures, which causes scarring, making subsequent draws difficult and painful. Novel blood collection systems may decrease discomfort in patients experiencing repeat blood draws. Methods Oncology outpatients (n = 101; criteria excluded 12) were recruited to determine their preference for either of two blood collection systems, the 23-gauge standard BD Vacutainer Push Button Blood Collection Set (Standard Push Button system) or the 25-gauge BD Vacutainer UltraTouch Push Button Blood Collection Set (UltraTouch Push Button system). Subjects received two blinded, randomized blood draws, one with each device and just one device for each arm. Subjects subsequently rated their blinded preference for blood collection system. Specimen quality was assessed for each device with measurements for plasma hemoglobin (Shimadzu UV-1800 spectrophotometer, Shimadzu), lactate dehydrogenase, and potassium (Vitros 4600/5600 analyzer, Ortho Diagnostics). Results Preference for the 25-gauge UltraTouch Push Button system over the 23-gauge Standard Push Button system was significant (UltraTouch, n = 51; Standard n = 30; no preference, n = 8; P = 0.0196). Regarding sample quality, the 25-gauge UltraTouch Push Button system had significantly lower plasma hemoglobin (average 5.34 mg/dL) vs the 23-gauge Standard Push Button system (9.37 mg/dL; P Conclusion Subjects in an oncology clinic preferred phlebotomy with the 25-gauge UltraTouch Push Button system, and samples using this device had less hemolysis as assessed by plasma hemoglobin.

Bovine brucellosis is among the top five diseases primarily threatening both public health and livestock economy. Available data are limited to central and highland areas of the country leaving documented literature on the disease in cattle to be found hardly in pastoral and agropastoral regions of the country. As a result, the magnitude and extent of the disease remained to be investigated. A cross-sectional study design was conducted on local Afar cattle aged six months and above from February 2017 to January 2019 in selected districts of Afar region. Technically, study districts and kebeles were selected purposively whereas simple random sampling technique was applied to select cattle owners and individual animals for sample collection. An average of 8 ml whole blood was drawn of jugular vein into plain vacutainer tube using sterile needle. Using Thrusfield formula, a total of 420 blood samples were collected. The sera were tested by RBPT and CFT tests for detection of Brucella antibodies. Data were analyzed using Stata v14.0. Of the 420 sera tested by RBPT, 50 were positive for Brucella antibodies providing an overall animal level prevalence of 11.9% and those RBPT positive sera were further retested by specific and sensitive confirmatory CFT test and 24 of the retested samples had come positive for the disease providing an overall individual animal seroprevalence of 5.7% over the three districts. Of the 3 associated factors (sex, age, and district) considered, only sex had significantly associated ( P < 0.05 : 0.036 ) with the disease. To estimate the strength of sex impact, odds ratio was generated using bivariate and multivariate logistic regression analyses with 95% CI and P < 0.05 providing OR of 2.484 (1.061–5.815) and 2.514 (1.041–6.07), respectively. Hence, the computations revealed that male cattle were 2.484 and 2.514 times more likely at higher risk for the disease as compared to their female counterparts.

Objective: The aim of the present study was to determine the effect of storage time and temperature on laboratory results of routine clinical biochemistry analytes in sera from apparently healthy volunteers. Materials and Methods: Ten healthy volunteers were instructed to fast overnight and 10 ml of blood was collected from each subject without anticoagulant (in red capped vacutainer). Samples were allowed to clot at room temperature for 20 min, centrifuged and serum separated which was stored in various aliquots. Baseline analysis (“0” day values) of 18 analytes in serum of each subject was done without delay on the same day of collection. Other aliquots were stored at 0° C and 4±1° C with cover of aluminium foil to avoid exposure of direct light and analysed on 3, 7, 15 and 30 days. Results: Urea, uric acid, phosphorus, TG and HDL were stable till 7 days whereas ALP was stable till 15 days but SGOT was stable upto 30 days at both 0° C and 4±1° C. ALP, Amylase and urea were stable up to 30 days at 0° C temperature. All analytes showed significant variation on 3rd day which were stored in room temperature except calcium which was stable. But glucose, creatinine, inorganic phosphorus and potassium were least stable and should be determined within 48 hours at 4±1° C and 24 hours at 23±1° C for these analytes. Conclusion: Various routine biochemical analytes were stable using the storage conditions tested in this study at least up to 7 days in usual refrigerator. This evidence can be used in exceptional circumstances because processing of any analyte on the same day should be done for better reproducibility. We recommend that samples should be analysed in the laboratory within preferably 24 h of collection to ensure valid results with the turn-around time from sample drawing to reporting the analytical result could be shortened. Beyond all this, it is even very important and useful to check the reliability of technical and instrumental resources that the laboratory will use during the study because molecular alterations of the analytes due to variable storage conditions can cause misleading results.

Background and aim The study aimed to determine whether the MPs levels and platelet activation are affected by the COVID-19 infection in both malignant and non-malignant patients compared to healthy individuals and define their contribution to the COVID-19 associated coagulopathy and the relation of these MPs to other hematologic parameters. Patients and methods We recruited 23 malignant patients with reverse transcription polymerase chain reaction (RT-PCR) positive COVID-19, also, 19 COVID-19 non-malignant patients, and 20 healthy volunteers were also enrolled for comparison. Blood samples were collected from patients and healthy donors into 5 mL vacutainer tube containing 3.5% buffered sodium citrate solution for measurement of total microparticles (TMPs), platelet microparticles (PMPs), endothelial microparticles (EMPs), CD62 activated platelets, and CD41 platelet marker. Results COVID-19 malignant patients had significantly lower hemoglobin and platelets compared to COVID non-malignant ones, while they had significantly higher C-reactive protein, LDH, AST, Albunim, creatinine, and prognostic index (PI) compared to COVID-19 non-malignant patients. significant accumulations of TMPs, PMPs, EMPs, and activated platelets in COVID-19 affected patients compared to healthy controls. TMPs, and EMPs were significantly accumulated in COVID-19 malignant compared to COVID-19 non-malignant patients with no significant difference in PMPs between both. Conclusion Circulating MPs and activated platelets may be promising novel prognostic biomarkers capable of identifying potentially severe COVID-19 patients who require immediate care especially in cancer patients.

Background In 2020, the SARS-CoV-2 virus spread worldwide and infected more that 10 million people, causing more than 500,000 deaths worldwide. The infection has systemic effects on the respiratory and cardiovascular systems; thus, patients can present a variety of symptoms from asymptomatic to rapid deaths. In this paper, we present the first case of post-mortem SARS-CoV-2 molecular testing in Western part of Romania in a deceased with disseminated intravascular coagulation (DIC) and elevated D-dimer levels. Methods During the autopsy which took place at the Institute of Forensic Medicine from Timisoara, Romania, blood sample was collected in a vacutainer with EDTA and sent to the Laboratory of Forensic Genetics from Victor Babes University of Medicine and Pharmacy, Timisoara, Romania. Viral RNA extraction was performed automated on the Maxwell 48 RSC Extraction System (Promega, USA) using the Maxwell RSC Viral Total Nucleic Acid Purification kit (Promega, USA). After RNA extraction, the samples were amplified on a 7500 real-time PCR (Applied Biosystems, USA) using the genesig® Real-Time PCR Assay (Primer Design, UK). Results The molecular testing showed a cycle threshold value of 23.4 (1.2 x 106 copies/mL), indicating increased viral loads, which correlated with the laboratory analysis results, especially with D-dimer levels. Conclusions In cases of coagulopathy of SARS-CoV-2, patients in hospitals should be monitored closely for thrombosis development. Thus D-dimer can be used as prognostic marker in monitoring the evolution of SARS-CoV-2 infected patients.

Supplemental Digital Content is Available in the Text., Purpose: To compare 3 methods for creating ethylenediaminetetraacetic acid (EDTA) solution using readily available Vacutainer tubes for the treatment of band keratopathy. Methods: All 3 protocols used commercially available Vacutainer blood collection tubes coated with K2EDTA. An osmometer was used to measure and compare the concentration of EDTA created using 3 different protocols. The time required for preparation of the solution was measured and compared to evaluate its efficiency for everyday clinical use. In addition, volume of EDTA solution obtained was measured for method 1. The most promising protocol for clinical use was then used for treatment of a series of patients. Results: Average osmolarity was 532, 285, and 422 for methods 1, 2, and 3, respectively (ANOVA P < 0.01, all Tukey honestly significant difference P < 0.01). For the respective mixtures, average concentration was 65, 35, and 52 mg/mL, and average time to create solution was 189, 38, and 83 seconds (ANOVA P < 0.01, all Tukey honestly significant difference P < 0.01). The most promising, method 3, was found to be safe and effective in removing calcium from the corneal stroma in a series of 5 patients with 6 eyes treated. It also yielded 25% more solution for clinical use than method 1. Conclusions: Method 3 using a single 10-mL Vacutainer tube with 18 mg of K2-EDTA had the best balance of effective concentration of EDTA, time to preparation, and simplicity of methodology, when compared with previously published methods 1 and 2. It also yielded a greater final volume of solution.

Background: Patients with spontaneous intracerebral hemorrhage (ICH) have high mortality and morbidity rates; approximately one-third of patients with ICH experience hematoma expansion (HE). The spot sign is an established and validated imaging marker for HE. High-sensitivity C-reactive protein (hs-CRP) is an established laboratory marker for inflammation and secondary brain injury following ICH. Objective: To determine the association between the spot sign and hs-CRP, hematoma expansion, and clinical outcomes. Methods: Between December 2014 and September 2016, we prospectively recruited 1,964 patients with acute symptomatic ICH at 13 hospitals in Beijing, China. Next, we selected 92 patients within 24 h of the onset of symptoms from this cohort for the present study. ICH was diagnosed in the emergency room by non-contrast computed tomography (NCCT) scans. Follow-up scans were carried out within 48 h to evaluate patients for HE. Multidetector computed tomography angiography (MDCTA) was also used to identify spot signs. Blood samples were collected from each patient at admission in EDTA tubes (for plasma) or vacutainer tubes (for serum). hs-CRP values were determined by a particle-enhanced immunoturbidimetric assay in the laboratory at Beijing Tiantan Hospital, Capital Medical University. Patients were categorized into two groups according to their hs-CRP levels (hs-CRP

Introduction: The mitochondrial membrane potential (Δψ) and level of carbonylated proteins in mononuclear leukocytes of peripheral blood (ML) is of interest as a possible marker of systemic mitochondrial dysfunction in COPD. Aim: Evaluation of the indicators of secondary mitochondrial dysfunction of ML in smokers and ex-smokers with exacerbation of COPD. Methods: The study included 25 people. The control group consisted of 8 healthy volunteers. 17 patients with non-infectious exacerbation of COPD were divided into groups of comparable age: smokers (n=9) and ex-smokers (n=8). Blood was taken on the second day of hospitalization using the BD Vacutainer CPT. Δψ of ML was assessed using the JC-1 Assay Kit on a FC 500 flow cytometer using Cyflogic. The level of carbonylated proteins (CP) of ML was determined spectrophotometrically in terms of 106cells/ml [Fomina N.V., et al. Angiol Sosud Khir.2015;21(1):29-34]. Results: Δψ in ML of the control group was 0.318± 0.031 and was higher relative to the group of smokers (0.244±0.046;p=0.01). When comparing the Δψ of ML in the control group and ex-smokers (0.297±0.047), differences were not found. Smokers ML characterized by the higher content of CP (234[221;452]) relative to the ex-smokers (169[101;237];p=0.04). The CP level of ML in the control group (21[16;65]) was lower both in comparison with the group of smokers (p= 0.0006) and ex-smokers (p=0.005). Conclusions: Smokers are characterized by a pronounced decrease in Δψ of mitochondria of mononuclear leukocytes in exacerbation of COPD compared with ex-smokers. A high level of carbonylated proteins in smokers indicates a greater severity of oxidative stress in this group of patients.

Aims: (a) To evaluate the types and frequencies of preanalytical errors occurring in a tertiary care hematology diagnostic center and (b) To evaluate differences if any, across groups [outpatient data (OPD) vs inpatient data (IPD), type of test requested [complete blood count (CBC) vs coagulation] and laboratory (routine vs emergency). Settings and Design: A prospective study was conducted over a period of nine months (August 2017–April 2018) to address the above objectives. All samples received in the clinical hematology division of our institute were included in the analysis. Materials and Methods: Categories of preanalytical errors were defined. This included insufficient, clotted, diluted, and lipemic samples. Clerical errors such as wrong/absent sample labeling, requisition form-sample mismatch, and wrong vacutainer selection were also documented. IPD and OPD data, as well as data pertaining to samples sent for different tests [complete blood count (CBC)/coagulation] and in the routine and emergency laboratories, were segregated. Statistical Analysis Used: All errors in each category were recorded as numbers and corresponding percentages (proportions). The two-tailed z-test was applied to assess the significance of the difference in proportions across all groups. Statistical significance was kept at P < 0.05. Results: A total of 189,104 samples were received in the clinical hematology laboratory during the aforementioned period, out of which preanalytical errors were found in 4052 (2.14%) samples. Inadequate sample quantity (ISQ) comprised the bulk of preanalytical errors in our laboratory (1.11% of total samples) followed by sample clots (0.88%). There was no significant difference in the error frequencies in OPD and IPD (P = 0.1031). The proportion of errors was higher in routine vis-à -vis emergency samples and also in samples sent for coagulation analysis vis-à -vis CBC.

BackgroundThe methylation pattern of cfDNA, isolated from liquid biopsies, is gaining substantial interest for diagnosis and monitoring of diseases. We have evaluated the impact of type of blood collection tube and time delay between blood draw and plasma preparation on bisulfite-based cfDNA methylation profiling.Methods15 tubes of blood were drawn from three healthy volunteer subjects (BD Vacutainer K2E EDTA spray tubes, Streck Cell-Free DNA BCT tubes, PAXgene Blood ccfDNA tubes, Roche Cell-Free DNA Collection tubes and Biomatrica LBgard blood tubes in triplicate). Samples were either immediately processed or stored at room temperature for 24 or 72 hours before plasma preparation. DNA fragment size was evaluated by capillary electrophoresis. Reduced representation bisulfite sequencing was performed on the cell-free DNA isolated from these plasma samples. We evaluated the impact of blood tube and time delay on several quality control metrics.ResultsAll preservation tubes performed similar on the quality metrics that were evaluated. Furthermore, a considerable increase in cfDNA concentration and the fraction of it derived from NK cells was observed after a 72-hour time delay in EDTA tubes.ConclusionThe methylation pattern of cfDNA is robust and reproducible in between the different preservation tubes. EDTA tubes processed as soon as possible, preferably within 24 hours, are the most cost effective. If immediate processing is not possible, preservation tubes are valid alternatives.

Caiman zoo breeding (Caiman crocodilus fuscus) has been developing with greater force in Colombia since the 90s. It is essential to evaluate the physiological ranges of the species to be able to assess those situations in which their health is threatened. The objective of the present study was to determine the typical hematological values of the Caiman (Caiman crocodilus fuscus) with the aid of the microhematocrit, the cyanmethemoglobin technique, and a hematological analyzer. The blood samples were taken from 120 young animals of both sexes in good health apparently (males 44 and females 76). The samples were taken from the coccygeal vein between the second and third interscalar space with lilac lid vacutainer tube whit acid ethylene-diamine-tetra-acetic as an anticoagulant. The animals are located in the middle of the Magdalena area in Colombia. The results obtained on average were Hematocrit: 23.36 %, Hemoglobin: 7.78 g/dl, Red blood cells: 3.89 106/µL, Average corpuscular volume: 60 fl, Average corpuscular hemoglobin concentration: 33.33 g/dl and Average corpuscular hemoglobin: 73.9 pg. This is the first study of hematological values for this particular species in Colombia. RESUMEN La zoocría de babilla (Caiman crocodrilus fuscus) se ha venido desarrollando con mayor fuerza en Colombia desde los años 90. Es muy importante evaluar los rangos fisiológicos de la especie para poder valorar aquellas situaciones en las que su salud se vea comprometida. El objetivo del presente trabajo fue determinar los valores hematológicos normales del caimán común (Caimán crocodrilus fuscus) con la ayuda del microhematocrito, la técnica de cianometahemoglobina y un analizador hematológico. Las muestras de sangre se obtuvieron de 120 animales en aparente buen estado de salud, machos (44) y hembras (76); se tomaron de la vena coccígea entre el segundo y tercer espacio interescamal con tubo vacutainer tapa lila, con ácido etilen-diamino-tetra-acético como anticoagulante. Los animales están ubicados en la zona del Magdalena medio en Colombia, todos en cautiverio. Los resultados obtenidos en promedio fueron: Hematocrito: 23,36 %, Hemoglobina: 7,78 g/dl, Glóbulos rojos: 3,89 106/µL, Volumen corpuscular medio: 60 fl, Concentración de Hemoglobina corpuscular media: 33,33 g/dl y Hemoglobina corpuscular media: 73,9 pg. Este es el primer reporte de valores hematológicos para esta especie en Colombia y Latinoamérica.

Dried blood spots (DBS) have proven to be a powerful sampling and storage method for newborn screening and many other applications. However, DBS methods have not yet been optimized for broad-spectrum targeted metabolomic analysis. In this study, we developed a robust, DBS-based, broad-spectrum, targeted metabolomic method that was able to measure over 400 metabolites from a 6.3 mm punch from standard Whatman 903TM filter paper cards. The effects of blood spot volumes, hematocrit, vacutainer chemistry, extraction methods, carryover, and comparability with plasma and fingerstick capillary blood samples were analyzed. The stability of over 400 metabolites stored under varying conditions over one year was also tested. No significant impacts of blood volume and hematocrit variations were observed when the spotted blood volume was over 60 &micro, L and the hematocrit was between 31% and 50%. The median area under the curve (AUC) of metabolites in the DBS metabolome declined by 40% in the first 3 months and then did not decline further for at least 1 year. All originally detectable metabolites remained within detectable limits. The optimal storage conditions for metabolomic analysis were &minus, 80 &deg, C with desiccants and without an O2 scavenger. The method was clinically validated for its potential utility in the diagnosis of the mitochondrial disease mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS). Our method provides a convenient alternative to freezing, storing, and shipping liquid blood samples for comparative metabolomic studies.

Introduction: The most common reason for rejection of blood samples in laboratory is because they are under-filled. Clinical and Laboratory Standards Institute (CLSI) guidelines state that blood vacutainers that contain either 10% higher or lower blood than the recommended volume must be rejected. Repeated phlebotomy has to be performed for such samples which can delay the turnaround time. Aim: To study the effects of underfilled K2EDTA vacutainers on automated haematology indices and compare them with the standard. Materials and Methods: This observational and comparative study was conducted at the department of pathology, Chettinad Hospital and Research Institute, Tamil Nadu for a duration of 6 months from January 2019 to June 2019. A 100 paired samples of underfilled K2EDTA vacutainers were compared with the standard for Complete Blood Count (CBC) analysis using BECKMAN COULTER LH 780 analyser. Statistical analyses was done using Statistical Package for Social Science (SPSS) software v.18 and paired student’s t-test. The p-value

Introduction: Serum samples of haemodialysed patients collected through vascular access devices, e.g. central venous catheter (CVC) can contain residual heparin, which can cause incomplete clotting and consequently fibrinogen interference in serum protein electrophoresis (SPE). We hypothesized that this problem may be overcome by addition of thrombin and aimed to find a simple thrombin-based method for fibrinogen interference removal. Materials and methods: Blood samples of 51 haemodialysed patients with CVC were drawn through catheter into Clot Activator Tube (CAT) and Rapid Serum Tube Thrombin (RST) vacutainers (Becton Dickinson, New Jersey, USA) following the routine hospital protocols and analysed with gel-electrophoresis (Sebia, Lisses, France). Samples were redrawn in the CAT tubes and re-analysed after being treated with thrombin using two methods: transferring CAT serum into RST vacutainer and treatment of CAT serum with fibrinogen reagent (Multifibren U, Siemens, Marburg, Germany). Results: Direct blood collection in RST proved to be slightly more efficient than CAT in removing the interfering band in beta fraction (CAT removed 6/51 and RST removed 12/51, P = 0.031). Transferring CAT serum into the RST vacutainer proved to be more efficient for subsequent removal of interfering band from CAT serum than the addition of fibrinogen reagent (39/45 vs. 0/45 samples with efficiently removed interfering band, P < 0.001). Conclusion: Fibrinogen interference caused by incomplete clotting because of residual heparin can be overcome by addition of thrombin. Transferring CAT serum into the RST vacutainer was the most efficient method.

Background Filling of citrate tubes with appropriate amount of blood is essential for obtaining reliable results of coagulation testing. This study aimed to verify whether results of routine coagulation tests are comparable when the new Becton Dickinson Vacutainer® Citrate Plus tubes are filled at minimum or optimal volume. Methods The study population consisted of 133 patients (40 on oral anticoagulant therapy), who had blood collected for routine coagulation testing. Two sequential Vacutainer® Citrate Plus tubes of the same type and lot were drawn. The first tube was collected after a butterfly needle was inserted into the vein, so that the air in the tubing was aspirated into the tube before blood (minimum fill volume), whilst the second was drawn at optimal fill volume. Experiments were repeated using 2.7-mL (n = 86) and 1.8-mL (n = 47) tubes. Results Prothrombin time (PT) and fibrinogen values were slightly but significantly decreased in tubes with minimum than in those with optimal fill volume. The activated partial thromboplastin time (APTT) was slightly prolonged in tubes with minimum than in those with optimal fill volume, but the difference was not statistically significant. An identical trend was noted in separate analyses for the 2.7-mL and 1.8-mL tubes. Spearman’s correlations between the two fill volumes were always >0.94 and bias was always within the quality specifications. Conclusions Blood drawing into Vacutainer® Citrate Plus tubes at minimum fill volume does not clinically bias routine coagulation testing.

Background: For blood, most 24/7 standard (immuno)chemistry parameters are either measured in serum or in lithium heparin plasma. Standard serum and plasma gel tubes have their shortcomings when timely analysis of high quality results is required. Serum requires clotting time and interference of gel globules in the plasma and adsorption of hydrophobic analytes into the gel layer potentially compromises high quality results from lithium heparin gel tubes. We sought to evaluate the impact of BD Vacutainer® Barricor™ Tube (Barricor™) on laboratory efficiency by measuring its effect on TAT and sample quality, as well as evaluate potential cost opportunities resulting from improved sample quality. Methods: TAT data and remediation activities were extracted and captured during two 6 months phases. Serum was used as the predominant matrix in the first phase and Barricor™ plasma was used in the second phase. Results: Barricor™ significantly reduced the median TAT, especially for routine-priority samples during peak-hours. The TAT key-performance-indicator (percentage of results available within 90 ​min) improved to >90% for STAT as well as routine priority samples. Converting from serum gel, Barricor™ reduced fibrin-related remediation activities from 2.3% to 0.4%. This resulted in remediation-related cost reduction of €6.010,47 over the study period. Conclusions: By implementing Barricor™, we saw a significant reduction in TAT and a reduction in fibrin-related remediation time and costs, when compared to a predominant serum workflow. The improved TAT opens up the possibility of consolidating to one single priority level, eliminating the need for the use of the STAT priority level. Keywords: Serum, Lithium heparin plasma, BD barricor, Turn-around-time, Remediation

Background An interlaboratory study was performed to compare the performance of the 3 mL TransFix®/EDTA Vacuum Blood Collection Tube (TVT), with the 5 mL Cyto-Chex® BCT tube (BCT). Both devices are intended for collection and storage of whole blood specimens for immunophenotyping of leukocytes by flow cytometry for up to 14 days. Methods One site in the United States and two in the United Kingdom tested samples from 10 HIV positive patients and four healthy subjects for a total of 42 samples. From each subject, three blood samples were collected: a BD 4 mL K3 EDTA Vacutainer (Vacutainer), a TVT, and a BCT. At all sites, samples were analyzed on a BD FACS Canto II flow cytometer for a full lymphocyte subset count within 6 h of collection (all devices) and on Day 11 and Day 15 (TVT and BCT only). Data obtained from the Vacutainer were used as the control data set with which TVT and BCT data were statistically compared. Results and conclusions Statistical concordance was demonstrated for both devices in relation to cell absolute count recovery. For cell marker signal, both devices exhibited a significant decrease in mean fluorescence intensity (MFI) for the detection of lymphocyte subsets and their target markers. There was a marked increase in autofluorescence observed for BCT stabilized lymphocytes whereas values for the TVTs were comparable to the control. There were eight instances of statistical equivalence between the level of antibody autofluorescence observed in the control tube and the TVT across both patient cohorts, versus two for the BCT. © 2018 International Clinical Cytometry Society.

Backgrounds The BD Vacutainer® Barricor™ Plasma collection tube (BD Barricor) uses an innovative non-gel separation method. This study compared the plasma residual cell count (PRCC) obtained from BD Barricor and from BD PST II plasma tubes. Methods Four BD Barricors and one BD PST II were collected from 40 donors. BD PST II was centrifuged at 1300g/10 min, while the BD Barricors were centrifuged at 1800g/10 min, 4000g/3 min, 4000g/7 min and 4000g/15 min. PRCC was evaluated measuring white blood cells (WBC), red blood cells (RBC) and Platelets (PLT) counts by Siemens ADVIA 2120. Cell-free hemoglobin was quantified by haemolysis index (HI) by Roche Cobas c501. Results BD PST II Median WBC, RBC and PLT counts were 0.38 (109/L), 0.0291 (1012/L) and 113.5 (109/L), respectively. Considering the BD PST II as reference, PRCC differences were expressed as median bias percentage. WBC showed a significant reduction at all the conditions (p Conclusions BD Barricors plasma quality improved with increasing the centrifugation times but already at 4000g/3 min, the suggested centrifugation condition, a significant improvement was achieved.