Optimization of specific adsorption conditions for hemoglobin in blood by graphite-phase carbon nitride

doi:10.12307/2023.546

Chinese Journal of Tissue Engineering Research ›› 2023, Vol. 27 ›› Issue (30): 4817-4823.doi: 10.12307/2023.546

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Optimization of specific adsorption conditions for hemoglobin in blood by graphite-phase carbon nitride

Lu Tian1, 2, Tao Yuan3, Yang Tuo1, 2, Sun Lulu2, Pei Xinyang1, Zhao Liang1, 2, Li Wenbin1, 4

1Department of Cardiothoracic Surgery, The Third Affiliated Hospital of Xinxiang Medical University, Xinxiang 453003, Henan Province, China; 2College of Life Science and Technology, Xinxiang Medical University, Xinxiang 453003, Henan Province, China; 3Department of Anesthesiology and Perioperative Medicine, People’s Hospital of Zhengzhou University, Zhengzhou 450003, Henan Province, China; 4Beijing Anzhen Hospital, Capital Medical University, Beijing 100029, China

Received:2022-07-25 Accepted:2022-09-21 Online:2023-10-28 Published:2023-04-03
Contact: Li Wenbin, Chief physician, Professor, Doctoral supervisor, Department of Cardiothoracic Surgery, The Third Affiliated Hospital of Xinxiang Medical University, Xinxiang 453003, Henan Province, China; Beijing Anzhen Hospital, Capital Medical University, Beijing 100029, China Zhao Liang, Associate professor, Master’s supervisor, Department of Cardiothoracic Surgery, The Third Affiliated Hospital of Xinxiang Medical University, Xinxiang 453003, Henan Province, China; College of Life Science and Technology, Xinxiang Medical University Xinxiang 453003, Henan Province, China
About author:Lu Tian, Master candidate, Department of Cardiothoracic Surgery, The Third Affiliated Hospital of Xinxiang Medical University, Xinxiang 453003, Henan Province, China; College of Life Science and Technology, Xinxiang Medical University, Xinxiang 453003, Henan Province, China
Supported by:
The Key Science and Technology Program of Henan Province, China, No. 182102310067 (to LWB); the Natural Science Foundation of Henan Province, No. 202300410311 (to ZL); a grant from Henan Fangyuan Medical Union Medical Equipment Co., No. J00043 (to ZL)

Abstract

Abstract: BACKGROUND: Currently, there has been no effective strategy for prolonging the 6-hour safe time limit of cardiopulmonary bypass mainly due to the accumulation of free hemoglobin.
OBJECTIVE: To investigate the optimum conditions and the highest adsorption efficiency of graphite-phase carbon nitride (g-C3N4) adsorbing free hemoglobin.
METHODS: Different concentrations of hemoglobin solutions were prepared to study the adsorption capacity of g-C3N4 for hemoglobin, and then the adsorption efficiency of characteristic proteins (albumin, hemoglobin, cytochrome C) in whole blood at different pH values was explored. Based on clinical application conditions, the influences of pH, g-C3N4: blood sample (mg/mL), reaction temperature (°C) and reaction time (minite) on adsorption efficiency were investigated. Using the model created by the response surface analysis software Design-Expert.8.0.6, pH (A), g-C3N4: blood sample (mg/mL) (B), reaction time (C), and reaction temperature (D) were set as independent variables, and the adsorption efficiency of free hemoglobin in whole blood as dependent variables, the experiment with four factors and three levels was designed. According to the single factor experiment results, the zero level and fluctuation zone of three factors were selected, and the absorbance was measured by ultraviolet spectrophotometer to calculate the content of free hemoglobin before and after adsorption, and then the adsorption efficiency was calculated. Finally, the specific adsorption effect of g-C3N4 on hemoglobin in whole blood was verified by electrophoresis.

RESULTS AND CONCLUSION: (1) With the increase of hemoglobin concentration in blood, the adsorption capacity of g-C3N4 to free hemoglobin increased gradually, but did not satisfy the linear relationship. (2) Single factor experiment showed that when the pH was 8.0, the adsorption efficiency of free hemoglobin in whole blood was 93.5%, and the adsorption of other proteins was few. When the pH value increases further, the adsorption efficiency for free hemoglobin initially remains stable and then gradually decreases. With the increase of g-C3N4: blood sample ratio, reaction time and reaction temperature, the adsorption efficiency of free hemoglobin in whole blood increased gradually. (3) The multiple regression equation of this experiment was calculated by Design-Expert.8.0.6 software, and the optimal action values of the four factors were obtained. The highest adsorption efficiency of free hemoglobin in whole blood was obtained by software analysis under the following experimental conditions: pH=7.5, in g-C3N4: blood sample (mg/mL) was 6.00:1, reaction temperature was 36.5 °C, reaction time was 60 minutes, and the adsorption efficiency of free hemoglobin in whole blood was 54.34%. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed that g-C3N4 achieved specific adsorption of hemoglobin in whole blood under the optimum adsorption conditions. (4) These results indicate that g-C3N4 can specifically adsorb free hemoglobin and improve the adsorption efficiency under the optimal adsorption conditions, which is expected to be a new biomedical material and further promote the development of tissue engineering materials.

Key words: graphite carbon nitride, free hemoglobin, specific adsorption, response surface analysis, optimum adsorption condition

CLC Number:

Lu Tian, Tao Yuan, Yang Tuo, Sun Lulu, Pei Xinyang, Zhao Liang, Li Wenbin. Optimization of specific adsorption conditions for hemoglobin in blood by graphite-phase carbon nitride[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(30): 4817-4823.

Figures/Tables 9

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Optimization of specific adsorption conditions for hemoglobin in blood by graphite-phase carbon nitride

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