Title Page
Abstract
Contents
LIST OF ABBREVIATIONS 15
PART Ⅰ. In Vitro Evaluation of DINCH-Plasticized Blood Bags for Red Blood Cell Storage with CPDA-1 Anticoagulant 17
1. INTRODUCTION 18
2. MATERIALS AND METHODS 21
2.1. Donor Recruitment 21
2.2. Blood Collection and Component Production 24
2.3. Blood Storage, Sample Preparation, and in vitro Testing Schedule 28
2.4. RBC Concentrate Quality Analysis 31
2.5. Statistical Analysis 41
3. RESULTS 42
3.1. Donor Recruitment and Study Progress 42
3.2. Component Integrity 44
3.3. RBC Concentrate Quality 44
4. DISCUSSION 90
PART Ⅱ. Analysis of DEHP and DINCH Concentrations in Blood Donors and Red Blood Cell Concentrates 100
1. INTRODUCTION 101
2. MATERIALS AND METHODS 103
2.1. Reagents 103
2.2. Calibration Standard Preparation 103
2.3. Sample Preparation 105
2.4. UHPLC-MS/MS Analysis 105
2.5. Evaluation of Analytical Performance 108
2.6. Identification of Plasticizer Contamination 108
2.7. Plasticizer Measurement in Donors and Blood Bags 108
2.8. Statistical Analysis 108
3. RESULTS 110
3.1. UHPLC MRM Chromatogram 110
3.2. Analytical Performance 111
3.3. Identification of Plasticizer Contamination 113
3.4. Plasticizer Measurement in Donors and Blood Bags 115
4. DISCUSSION 120
REFERENCES 127
CONFLICT OF INTEREST 135
국문 초록 136
Table 1. Inclusion and exclusion criteria of blood donation participants. 23
Table 2. In vitro testing plan schedule throughout the study course. 30
Table 3. Information on reagent contents for 2,3-DPG measurement. 35
Table 4. Information on reagent contents for ATP measurement. 38
Table 5. Plasma dilution rates for the measurement of analytes using Vitros 5600. 40
Table 6. Characteristics of the donors that were qualified for study participation. 43
Table 7. Hemolysis (%) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA-1. 47
Table 8. Plasma hemoglobin (mg/dL) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA-1. 48
Table 9. Hemoglobin (g/dL) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA-1. 58
Table 10. Hematocrit (%) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA- 1. 59
Table 11. RBC count (×10⁶/μL) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA-1 60
Table 12. WBC count (×10³/μL) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA-1. 61
Table 13. Platelet count (×10³/μL) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA-1. 62
Table 14. MCV (fL) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA-1. 63
Table 15. MCH (pg) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA-1. 64
Table 16. MCHC (g/dL) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA- 1. 65
Table 17. EMA binding test (%) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA-1. 68
Table 18. 2,3-DPG (μmol/g Hb) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA-1. 72
Table 19. ATP (μmol/g Hb) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA-1. 73
Table 20. pH results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA-1. 78
Table 21. pCO₂ (mmHg) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA- 1. 79
Table 22. pO₂ (mmHg) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA- 1. 80
Table 23. Glucose (mg/dL) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA-1. 86
Table 24. Lactate (mmol/L) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA-1. 87
Table 25. K⁺ (mmol/L) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA- 1. 88
Table 26. Na⁺ (mmol/L) results for matched quintets (n=20) of RBC concentrates stored in DEHP and DINCH bags with CPDA- 1. 89
Table 27. Chemical structure and molecular weight of plastic materials. 104
Table 28. UHPLC-MS/MS settings and MRM transition information. 107
Table 29. Results of within-run precision for the measurement of DEHP and DINCH 111
Table 30. DEHP concentration (mg/L) results for matched quintets of RBC concentrates stored in DEHP and DINCH bags with CPDA-1. 118
Table 31. DINCH concentration (mg/L) results for matched quintets of RBC concentrates stored in DEHP and DINCH bags with CPDA-1 119
Figure 1. Advertisement poster to recruit blood donors for study participation. 22
Figure 2. Study design. A pool-and-split study was used to evaluate DINCH- plasticized blood bags for red blood cell storage with CDPA-1 anticoagulant. Whole... 27
Figure 3. Hemolysis of RBC concentrate units stored in DEHP- and DINCH- plasticized blood bags for 35 days (n=20). (A) The DINCH bags showed increased... 45
Figure 4. Median values of plasma hemoglobin levels of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). The... 46
Figure 5. Median values of hemoglobin levels of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 50
Figure 6. Median values of hematocrit levels of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 51
Figure 7. Median values of RBC count of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 52
Figure 8. Median values of WBC count of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 53
Figure 9. Median values of platelet count of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 54
Figure 10. Median values of MCV of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 55
Figure 11. Median values of MCH of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 56
Figure 12. Median values of MCHC of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 57
Figure 13. Median values of EMA binding test of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 67
Figure 14. Median values of 2,3-DPG of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 2,3-DPG was not measured... 70
Figure 15. Median values of ATP of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 71
Figure 16. Median values of pH of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 75
Figure 17. Median values of pCO₂ of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 76
Figure 18. Median values of pO₂ of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 77
Figure 19. Median values of glucose of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 82
Figure 20. Median values of lactate of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 83
Figure 21. Median values of K⁺ of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 84
Figure 22. Median values of Na⁺ of RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days (n=20). 85
Figure 23. Gradient information of UHPLC for DEHP and DINCH measurement. 106
Figure 24. Representative chromatogram for the measurement of DEHP and DINCH 110
Figure 25. Linearity for DEHP and DINCH measurement. 112
Figure 26. Comparison between blank and 50 mg/L DEHP solution chromatograms. 114
Figure 27. Median values of DEHP in pooled blood (day 0) and RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days. 116
Figure 28. Median values of DINCH in pooled blood (day 0) and RBC concentrate units stored in DEHP- and DINCH-plasticized blood bags for 35 days. 117