ISLH

Laboratory Hematology 4:1720 1998 Carden Jennings Publishing Co., Ltd.

#97-24.Leong

Official Publication

Performance of K 2EDTA- vs. K 3EDTA-collected blood specimens on various hematology analyzers

J. PHILLIPS,1 J. COINER,2 E. SMITH,3 D. BECKER,4 J. LEONG5
1 2

Kaiser Permanente Regional Reference Lab, Aurora, CO Culpeper Memorial Hospital, Culpeper, VA 3 Northeast Medical Center, Concord, NC 4 Vencor Hospital, Arlington, VA 5 Coulter Corporation, Miami, FL

ABSTRACT
This study compared the performance of specimens preserved with potassium (K2) ethylene diamine tetraacetic acid (EDTA) and K3EDTA anticoagulants when analyzed on hematology analyzers that use histogram differential technology. Four clinical institutions collected a total of 471 whole blood specimens for accuracy comparison; 30 specimens were collected for stability studies. The specimens were examined on one hematology analyzer at each site for complete blood count (CBC) and white blood cell differential results. Statistical analyses included mean percent differences, distribution of percent differences, standard deviation, coefcient of variation, analysis of variance, correlation coefcient, and regression. All data showed excellent agreement for CBC and differential results between the two salts of EDTA.
Lab Hematol 4:1720, 1998

ential technology. Other studies have shown little difference in anticoagulant types when specimens are collected, stored, and analyzed under proper conditions [2]. Because this study was designed to compare anticoagulant performance under routine laboratory conditions using histogram differential technology, however, possible variations in test quality, such as an improper ratio of blood to EDTA, were not evaluated.

MATERIALS AND METHODS

Each venous whole blood specimen was collected in both K 2EDTA and K 3EDTA (Vacutainer Plus collection tubes, Becton

TABLE 1. Accuracy study: CBC parameters of four test sites

Mean percent difference ([K3 K2]/K3 100)/n JT MDII ONYX STKR WBC 103 cells/L RBC 106 cells/L Hgb g/dL MCV fL PLT 103 cells/L 0.0 0.04 0.2 0.4 3 0.0 0.0 0.0 0.2 2 0.1 0.04 0.1 0.2 3 0.0 0.04 0.1 0.3 0.3

KEY WORDS:

Anticoagulant Blood collection Histogram differential

INTRODUCTION
In the clinical laboratory, plastic tubes containing dipotassium ethylene diamine tetraacetic acid (K 2EDTA) have emerged as an alternative to glass tubes containing tripotassium ethylene diamine tetraacetic acid (K 3EDTA). This study evaluated various hematological parameters of blood preserved with K 2EDTA as compared with that preserved with K 3EDTA. Previous studies have examined the accuracy and stability of results from instruments with volume, conductivity, scatter (VCS) technology for the white blood cell (WBC) differential [1], whereas this study examined results from instruments with histogram differ-

WBC, white blood cell; RBC, red blood cell; Hgb, hemoglobin; MCV, mean corpuscular volume; PLT, platelet.

TABLE 2. Accuracy study: differential parameters of four test sites

Mean percent difference ([K3 K2]/K3 100)/n JT MDII ONYX STKR LY% MO% GR% 0.5 0.2 0.5 0.3 0.5 0.2 0.2 0.1 0.0 0.1 0.5 0.5

This study was sponsored by Coulter Corporation, Miami, FL. Address correspondence to Judy Leong, MC 12-A03, Coulter Corporation, Miami, FL 33196-2500. Received 4 November 1997; accepted 8 January 1998

LY%, lymphocyte percent; MO%, monocyte percent; GR%, granulocyte percent.

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TABLE 3. Analysis of variance

Instrument Parameter WBC F ratio F crit p value Correlation coefcient RBC F ratio F crit p value Correlation coefcient Hgb F ratio F crit p value Correlation coefcient MCV F ratio F crit p value Correlation coefcient RDW F ratio F crit p value Correlation coefcient PLT F ratio F crit p value Correlation coefcient MPV F ratio F crit p value Correlation coefcient LY% F ratio F crit p value Correlation coefcient MO% F ratio F crit p value Correlation coefcient GR% F ratio F crit p value Correlation coefcient RDW, red cell distribution width; MPV, mean platelet volume. JT MDII ONYX STKR

0.0019 3.881 0.965 0.999 0.3217 3.881 0.5711 0.997 0.5636 3.881 0.4536 0.997 0.2768 3.881 0.599 0.986 0.7589 3.881 0.384 0.981 0.033 3.881 0.855 0.986 0.0196 3.881 0.888 0.975 0.0693 3.881 0.793 0.993 0.173 3.884 0.173 0.782 0.857 3.884 0.857 0.99

0.0002 3.889 0.989 0.992 0.0099 3.889 0.921 0.957 0.016 3.889 0.898 0.975 0.08 3.889 0.777 0.991 0.107 3.889 0.744 0.979 0.054 3.889 0.816 0.973 0.005 3.889 0.944 0.946 0.105 3.889 0.747 0.991 2.766 3.889 0.098 0.7 0.033 3.889 0.857 0.975

0.0019 3.884 0.965 0.9971 0.1399 3.884 0.7088 0.986 0.1053 3.884 0.7459 0.9823 0.0525 3.884 0.819 0.9927 0.0214 3.884 0.884 0.9944 0.567 3.884 0.812 0.9925 0.0047 3.884 0.946 0.9812 0.0103 3.884 0.9191 0.994 0.1867 3.884 0.666 0.786 0.00047 3.884 0.983 0.989

0.003 3.876 0.954 0.999 0.213 3.876 0.645 0.997 0.3811 3.876 0.537 0.991 0.166 3.876 0.683 0.994 0.389 3.876 0.844 0.993 0 3.876 0.982 0.997 0.295 3.876 0.587 0.979 0.002 3.88 0.968 0.997 2.98 3.883 0.086 0.775 0.058 3.883 0.81 0.995

Performance of K 2EDTA vs. K 3EDTA

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FIGURE 1. Stability study: mean difference from 1 hour (all sites).

Dickinson, Franklin Lakes, NJ) and analyzed on COULTER JT, MDII, ONYX, or STKR automated hematology instruments. Specimens obtained reect routine population distribution available at each of the four test sites. In the accuracy study, all specimens were analyzed within 30 minutes to 5 hours of collection. The K 2EDTA and K 3EDTA tubes for a single specimen were analyzed within 1 hour of each other. In the stability studies, specimens were analyzed at the following times after phlebotomy: 5 minutes, 15 minutes, 30 minutes, 1 hour (baseline), 4 hours, 8 hours, 12 hours (if available),

24 hours, and 30 hours. Between analyses, specimens were stored at ambient room temperature.

RESULTS AND DISCUSSION

Accuracy During the evaluation period at each site accuracy was assessed by analyzing a minimum of 100 specimens collected in K 2EDTA and K 3EDTA anticoagulants. Specimen results with incomplete computations were excluded. Results showed excellent agreement

FIGURE 2. Stability study: change over time in red cell distribution width (RDW).

FIGURE 3. Stability study: change over time in mean corpuscular volume (MCV).

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TABLE 4. Mean percent differences for all four sites

Mean percent difference ([K 3 K 2]/K 3 100)/n WBC 103 cells/L RBC 106 cells/L Hgb g/dL MCV fL PLT 103 cells/L 0.0% 0.4% 0.6% 0.3% 0.6%

for complete blood count (CBC) and differential parameters; mean percent differences are listed in Tables 1 and 2. One-way analysis of variance (ANOVA) for CBC and differential parameters was evaluated for statistical signicance. Variance between samples was compared with that within samples (F ratio) for the two anticoagulants. The F ratios for CBC and differential parameters were not close to the critical F value (F crit), indicating that there is no significant difference among the sample means. The p value for CBC and differential parameters was greater than 0.05, indicating that the means from the two anticoagulant populations are equivalent (Table 3). Correlation coefcients for all parameters except MO% were 0.946 or better (Table 3). Specimen Stability Stability studies were designed to compare the time frame in which specimens yielded acceptable results over a 30-hour period. Results from 30 specimens were reviewed for numerical and agging consistency. Data for K 2EDTA-preserved specimens were compared with data for K 3EDTA-preserved specimens measured at the same times. Sample results were averaged, and differences between samples collected and stored in the two anticoagulants were calculated. Figure 1 shows trending after 8 hours, including an increase in the granulocyte percentages and a decrease in the lymphocyte and

monocyte percentages. It is important to note, however, that both K 2EDTA and K 3EDTA samples behaved similarly. In earlier studies by Brittin et al. [3], slight swelling of red blood cells (RBCs) occurred when specimens were stored at room temperature, thus resulting in an increased mean corpuscular volume (MCV) over time. Goosens et al. [2] demonstrated an increased red cell distribution width (RDW) for specimens stored in EDTA salts. As seen in Figures 2 and 3, these trends were confirmed by this study; the RDW and MCV showed an upward trend for both K 2EDTA and K 3EDTA after the 8-hour time point. This study indicated that the COULTER JT, MDII, ONYX, or STKR automated hematology instrument results for CBC and automated differential parameters obtained from specimens collected in K 2 EDTA were similar to those obtained using K 3 EDTA. Unlike the studies conducted by Koepke et al. in which a dilution effect was seen with liquid K 3EDTA tubes [4], this study agreed with results obtained by Brunson et al. [1] in that a dilution effect was not apparent. This result is illustrated in Table 4 in which the mean percent difference between the anticoagulants is shown for the directly measured CBC parameters. Equivalence in the use of these two anticoagulants in a clinical laboratory environment is supported by this study.

REFERENCES
1 BRUNSON D, SMITH D, BAK A, PRZYK E, SHERIDAN B, MUNCER D: Comparing hematology anticoagulants: K 2EDTA vs. K 3EDTA. Lab Hematol 1:112, 1995 2 GOOSENS W, VAN DUPPEN V, VERWILGHEN RL: K 2 or K 3EDTA: The anticoagulant of choice in routine hematology? Clin Lab Haematol 13:291, 1991 3 BRITTIN GM, BRECHER G, JOHNSON C, ELASHOFF R: Stability of blood in commonly used anticoagulants. Am J Clin Pathol 52:690, 1969 4 KOEPKE JA, VAN ASSENDELFT OW, BULL BS, RICHARDSON-JONES A: Standardization of EDTA anticoagulation for blood counting procedures. Labmedica 5:15, 1988/1989