Citation: ZHOU Lang, LI Qiao-Jing, LI Yong-Sheng, GAO Xiu-Feng. Rapid Determination of Pyruvic Acid in Erythrocytes by Lysing-heating Fluorescence Capillary Method[J]. Chinese Journal of Analytical Chemistry, ;2012, 40(11): 1725-1729. doi: 10.3724/SP.J.1096.2012.20271 shu

Rapid Determination of Pyruvic Acid in Erythrocytes by Lysing-heating Fluorescence Capillary Method

1.
¹ School of Chemical Engineering, Sichuan University, Chengdu 610065, China;

Corresponding author: LI Yong-Sheng,
Received Date: 9 March 2012
Available Online: 25 May 2012

In the conventional method of erythrocytes treatment, there was the efflux of pyruvic acid in erythrocytes (PAE) during washing and separation of erythrocytes/leukocytes using the physiological saline. This phenomenon caused the deviation of the measured value of PAE from its real concentration. This research designed a method comprising the steps of employing the erythrocyte lysis buffer to specifically lyse the red blood cells, separating leukocytes and platelets by centrifugation, removing the protein by heating the hemolysate, and detecting PAE using the pyruvate-enzyme fluorescence capillary analysis. The optimized procedures for the blood pretreatment were as follows: at the centrifugal speed of 16000 r/min, the blood sample was centrifuged for 1 min to obtain erythrocytes, after the erythrocytes were lysed by the lysis buffer, the hemolysate was heated for 5 min at 100℃. Through the contrast experiment, it was found that this method was superior to other erythrocyte treatment method. The pyruvate determination method in this research had a good linearity in the range of 10-120 mmol/L, a detection limit of 0.98 mmol/L, and a sensitivity of 5.64 F L/mmol, it was 60 times higher than that of references method. This method offered a relative standard deviation of PAE less than 2.8% (n=11), and a recovery range of 98.3%-104.1%.
- Enzyme fluorescence capillary analysis,
- Pyruvic acid,
- Erythrocytes,
- Lysing-heating method
1. [1]
  1 Lam T K T, Gutierrez-Juarez R, Pocai A, Rossetti L. Science, 2005, 309(5736): 943-947
2. [2]
  2 Gajovic N, Binyamin G, Warsinke A, Scheller F W, Heller A. Anal. Chem., 2000, 72(13): 2963-2968
3. [3]
  3 Horiguchi N, Horiguchi H. E. U. Patent, 1522852, 2003
4. [4]
  4 Shepotinovskii V I, Barsukova O I, Mikarshinovskii Z I. S. U. Patent, 1770906, 1972
5. [5]
  5 Darghouth D, Koehl B, Heilieret J F, Madalinski G, Bovee P, Bosman G, Delaunay J, Junot C, Roméo P H. Haematol.-Hematol. J., 2011, 96(12): 1861-1865
6. [6]
  6 Markuszewski M J, Szczykowska M, Siluk D, Kaliszan R. J. Pharm. Biomed. Anal., 2005, 39 (3-4): 636-642
7. [7]
  7 Ogasawara Y, Funakoshi M, Ishii K. Blood Cell Mol. Dis., 2008, 41(3): 237-243
8. [8]
  8 LI Yong-Sheng, GAO Xiu-Feng. Spectrosc. Spect. Anal., 2007, 27(8): 1565-1569
9. [9]
  李永生, 高秀峰. 光谱学与光谱分析, 2007, 27(8): 1565-1569
10. [10]
  9 Li Y S, Gao X F. Anal. Chim. Acta, 2007, 588 (1): 140-146
11. [11]
  10 GAO Xiu-Feng, LI Yong-Sheng, JIANG Yan-Xia. Chinese J. Anal. Chem., 2006, 34(9): 1220-1222
12. [12]
  高秀峰, 李永生, 姜艳霞. 分析化学, 2006, 34(9): 1220-1222
13. [13]
  11 Zhao Y Y, Gao X F, Li Y S, Ju X, Zhang J, Zheng J. Talanta, 2008, 76(2): 265-270
14. [14]
  12 Li Y S, Du Y D, Chen T M, Gao X F. Biosens. Bioelectron., 2010, 25(6): 1382-1388
15. [15]
  13 LI Yong-Sheng, YANG Wei, LI Qiao-Jing, ZHOU Lang, GAO Xiu-Feng. Chinese J. Anal. Chem., 2011, 39(7): 1058-1064
16. [16]
  李永生, 杨微, 李乔婧, 周朗, 高秀峰. 分析化学, 2011, 39(7): 1058-1064
17. [17]
  14 LI Yong-Sheng, GAO Xiu-Feng. China Patent, 1737538, 2012
18. [18]
  李永生, 高秀峰. 中国专利, 1737538, 2012
19. [19]
  15 Hawley T S, Hawley R G. Flow Cytometry Protocols. 2nd ed., New York: Humana Press, 2004: 164
20. [20]
  16 Dubinsky W P, Racker E. J. Membr. Biol., 1978, 44(1): 25-36
21. [21]
  17 ZHU Zhong-Yong. Practical Medical Laboratory Sciences. Beijing: People's Military Medical Press, 1992: 436-440
22. [22]
  朱中庸. 实用医学检验学. 北京: 人民军医出版社, 1992, 436-440
1. [1]
  Siyi ZHONG , Xiaowen LIN , Jiaxin LIU , Ruyi WANG , Tao LIANG , Zhengfeng DENG , Ao ZHONG , Cuiping HAN . Targeting imaging and detection of ovarian cancer cells based on fluorescent magnetic carbon dots. Chinese Journal of Inorganic Chemistry, 2024, 40(8): 1483-1490. doi: 10.11862/CJIC.20240093
2. [2]
  Yuan Zheng , Quan Lan , Zhenggen Zha , Lingling Li , Jun Jiang , Pingping Zhu . Teaching Reform of Organic Synthesis Experiments by Introducing Reverse Thinking and Design Concepts: Taking the Synthesis of Cinnamic Acid Based on Retrosynthetic Analysis as an Example. University Chemistry, 2024, 39(6): 207-213. doi: 10.3866/PKU.DXHX202310065
3. [3]
  Yanxi LIU , Mengjia XU , Haonan CHEN , Quan LIU , Yuming ZHANG . A fluorescent-colorimetric probe for peroxynitrite-anion-imaging in living cells. Chinese Journal of Inorganic Chemistry, 2025, 41(6): 1112-1122. doi: 10.11862/CJIC.20240423
4. [4]
  Mi Wen , Baoshuo Jia , Yongqi Chai , Tong Wang , Jianbo Liu , Hailong Wu . Improvement of Fluorescence Quantitative Analysis Experiment: Simultaneous Determination of Rhodamine 6G and Rhodamine 123 in Food Using Chemometrics-Assisted Three-Dimensional Fluorescence Method. University Chemistry, 2025, 40(4): 390-398. doi: 10.12461/PKU.DXHX202405147
5. [5]
  Zhaoxin LI , Ruibo WEI , Min ZHANG , Zefeng WANG , Jing ZHENG , Jianbo LIU . Advancements in the construction of inorganic protocells and their cell mimic and bio-catalytical applications. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2286-2302. doi: 10.11862/CJIC.20240235
6. [6]
  Shuyu Liu , Xiaomin Sun , Bohan Song , Gaofeng Zeng , Bingbing Du , Chongshen Guo , Cong Wang , Lei Wang . Design and Fabrication of Phospholipid-Vesicle-based Artificial Cells towards Biomedical Applications. University Chemistry, 2024, 39(11): 182-188. doi: 10.12461/PKU.DXHX202404113
7. [7]
  Liqiang Huang , Peng Lin . 数-图分析法解释仪器分析实验课程教学中的难点. University Chemistry, 2025, 40(6): 353-359. doi: 10.12461/PKU.DXHX202407074
8. [8]
  Shipeng WANG , Shangyu XIE , Luxian LIANG , Xuehong WANG , Jie WEI , Deqiang WANG . Piezoelectric effect of Mn, Bi co-doped sodium niobate for promoting cell proliferation and bacteriostasis. Chinese Journal of Inorganic Chemistry, 2024, 40(10): 1919-1931. doi: 10.11862/CJIC.20240094
9. [9]
  Peng GENG , Guangcan XIANG , Wen ZHANG , Haichuang LAN , Shuzhang XIAO . Hollow copper sulfide loaded protoporphyrin for photothermal-sonodynamic therapy of cancer cells. Chinese Journal of Inorganic Chemistry, 2024, 40(10): 1903-1910. doi: 10.11862/CJIC.20240155
10. [10]
  Jiahao Zeng , Hui Chao . 诱导程序性细胞死亡的金属抗肿瘤药物研究. University Chemistry, 2025, 40(6): 145-159. doi: 10.12461/PKU.DXHX202406019
11. [11]
  Hongmei Chai , Yixia Ren , Xiangyang Hou , Long Tang , Jiawei Xie . 智能手机光传感的“丙酮碘化反应”实验改进. University Chemistry, 2025, 40(6): 193-200. doi: 10.12461/PKU.DXHX202407086
12. [12]
  Di WU , Ruimeng SHI , Zhaoyang WANG , Yuehua SHI , Fan YANG , Leyong ZENG . Construction of pH/photothermal dual-responsive delivery nanosystem for combination therapy of drug-resistant bladder cancer cell. Chinese Journal of Inorganic Chemistry, 2024, 40(9): 1679-1688. doi: 10.11862/CJIC.20240135
13. [13]
  Liwei Wang , Guangran Ma , Li Wang , Fugang Xu . A Comprehensive Analytical Chemistry Experiment: Colorimetric Detection of Vitamin C Using Nanozyme and Smartphone. University Chemistry, 2024, 39(8): 255-262. doi: 10.3866/PKU.DXHX202312094
14. [14]
  Xinyu ZENG , Guhua TANG , Jianming OUYANG . Inhibitory effect of Desmodium styracifolium polysaccharides with different content of carboxyl groups on the growth, aggregation and cell adhesion of calcium oxalate crystals. Chinese Journal of Inorganic Chemistry, 2024, 40(8): 1563-1576. doi: 10.11862/CJIC.20230374
15. [15]
  Yinwu Su , Xuanwen Zheng , Jianghui Du , Boda Li , Tao Wang , Zhiyan Huang . Green Synthesis of 1,3-Dibromoacetone Using Halogen Exchange Method: Recommending a Basic Organic Synthesis Teaching Experiment. University Chemistry, 2024, 39(5): 307-314. doi: 10.3866/PKU.DXHX202311092
16. [16]
  Qingying Gao , Tao Luo , Jianyuan Su , Chaofan Yu , Jiazhu Li , Bingfei Yan , Wenzuo Li , Zhen Zhang , Yi Liu . Refinement and Expansion of the Classic Cinnamic Acid Synthesis Experiment. University Chemistry, 2024, 39(5): 243-250. doi: 10.3866/PKU.DXHX202311074
17. [17]
  Lanjun Cheng , Xinyuan Wang , Jie An , Xiang Wu , Chengfeng Zhu , Yanming Fu , Yougui Li . Improvement of the Resolution Experiment of Racemic Tartaric Acid. University Chemistry, 2025, 40(7): 277-285. doi: 10.12461/PKU.DXHX202408010
18. [18]
  Zhou Fang , Zhihao Zhang , Weihan Jiang , Kin Shing Chan . Warfarin: From Poison to Cure, the Remarkable Journey of a Molecule. University Chemistry, 2025, 40(4): 326-330. doi: 10.12461/PKU.DXHX202406038
19. [19]
  Hexing SONG , Zan SUN . Synthesis, crystal structure, Hirshfeld surface analysis, and fluorescent sensing for Fe³⁺ of an Mn(Ⅱ) complex based on 1-naphthalic acid. Chinese Journal of Inorganic Chemistry, 2025, 41(5): 885-892. doi: 10.11862/CJIC.20240402
20. [20]
  Meijin Li , Xirong Fu , Xue Zheng , Yuhan Liu , Bao Li . The Marvel of NAD⁺: Nicotinamide Adenine Dinucleotide. University Chemistry, 2024, 39(9): 35-39. doi: 10.12461/PKU.DXHX202401027

Get Citation

PDF

XML

Metrics

PDF Downloads(0)
Abstract views(449)
HTML views(54)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142