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Method for detecting content of methylene blue in blood plasma

A technology of methylene blue and plasma, which is applied in the field of pharmaceutical analysis, bioanalysis and analytical chemistry, and clinical testing. It can solve problems such as method failure, low absorbance value, and large plasma consumption, and achieve high detection precision and accuracy. , The determination method is simple, and the effect of linear regression is good

Inactive Publication Date: 2012-10-03
苏州市中心血站 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in practical application, it is found that this method has two significant shortcomings: (1) the operation is particularly cumbersome, time-consuming, and consumes a large amount of plasma; (2) the absorbance value of the sample is extremely low, usually less than 0.01; this is consistent with the "People's Republic of China Compared with the absorbance 0.3~0.7 required for accurate determination of content under "Appendix IV A UV-Vis Spectrophotometry" in the Pharmacopoeia, the determined residual amount is obviously inaccurate
This will obviously lead to the invalidation of the method in Document 1, which cannot be used for routine detection of plasma methylene blue residues

Method used

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  • Method for detecting content of methylene blue in blood plasma

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Embodiment 1

[0039] A method for detecting methylene blue content in plasma, comprising the steps of:

[0040] Add 280 μL of carboxymethyl-β-cyclodextrin saturated solution to 300 μL of blank plasma, and add 120 μL of PBS buffer to obtain a plasma blank sample;

[0041] Add 56 μL of 5 μmol / L methylene blue stock solution to 300 μL blank plasma, add 280 μL of carboxymethyl-β-cyclodextrin saturated solution, and add 64 μL of PBS buffer solution to obtain a plasma standard sample containing methylene blue;

[0042] Add 280 μL of carboxymethyl-β-cyclodextrin saturated solution to 300 μL of the plasma sample to be tested containing methylene blue, and add 120 μL of PBS buffer to obtain the plasma sample to be tested;

[0043] A fluorescence spectrophotometer is used for detection, and the detection conditions are: excitation wavelength 665nm, emission wavelength 680nm, excitation and emission slit widths of 5nm, scanning speed 100nm / min, excitation optical path 10mm, emission optical path 10mm,...

Embodiment 2

[0049] A method for detecting methylene blue content in plasma, comprising the steps of:

[0050] Add 50 μL of 0.1 μmol / L methylene blue stock solution; 28 μL, 56 μL of 5 μmol / L methylene blue stock solution; 28 μL, 42 μL of 25 μmol / L methylene blue stock solution, and add carboxymethyl - Add 280 μL of β-cyclodextrin saturated solution to 70 μL, 92 μL, 64 μL, 92 μL and 78 μL of PBS buffer respectively to obtain 5 plasma standard samples containing methylene blue;

[0051] Add 280 μL of carboxymethyl-β-cyclodextrin saturated solution to 300 μL of the plasma sample to be tested containing methylene blue, and add 120 μL of PBS buffer to obtain the plasma sample to be tested;

[0052] A fluorescence spectrophotometer is used for detection, and the detection conditions are: excitation wavelength 665nm, emission wavelength 680nm, excitation and emission slit widths of 5nm, scanning speed 100nm / min, excitation optical path 10mm, emission optical path 10mm, measure the above sample T...

Embodiment 3

[0054] A method for detecting methylene blue content in plasma, comprising the steps of:

[0055] Add 280 μL of carboxymethyl-β-cyclodextrin saturated solution to 300 μL of blank plasma, and add 120 μL of PBS buffer to obtain a plasma blank sample;

[0056] Add 50 μL of 0.1 μmol / L methylene blue stock solution; 28 μL, 56 μL of 5 μmol / L methylene blue stock solution; 28 μL, 42 μL of 25 μmol / L methylene blue stock solution, and add carboxymethyl - Add 280 μL of β-cyclodextrin saturated solution to 70 μL, 92 μL, 64 μL, 92 μL and 78 μL of PBS buffer respectively to obtain 5 plasma standard samples containing methylene blue;

[0057] Add 280 μL of carboxymethyl-β-cyclodextrin saturated solution to 300 μL of the plasma sample to be tested containing methylene blue, and add 120 μL of PBS buffer to obtain the plasma sample to be tested;

[0058] A fluorescence spectrophotometer is used for detection, and the detection conditions are: excitation wavelength 665nm, emission wavelength 6...

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Abstract

The invention discloses a method for detecting the content of methylene blue in blood plasma. The method comprises the following steps of: (a) adding carboxymethyl-beta-cyclodextrin saturated solution into blank blood plasma, enabling the concentration of the mixed solution to reach 1-4.5mmol / L and maintaining the constant volume to obtain a blood plasma blank sample; (b) adding quantitative methylene blue solution into the blank blood plasma and adding the carboxymethyl-beta-cyclodextrin saturated solution to obtain a standard blood plasma sample containing the methylene blue with the concentration of Cs; and (c) adding the carboxymethyl-beta-cyclodextrin saturated solution in a blood plasma sample containing the methylene blue to obtain a blood plasma to-be-measured sample containing the methylene blue with the concentration of Cs; detecting the strength or the area of a fluorescence peak of the sample; and then calculating according to a reference substance comparison method to obtain the concentration Cs of the methylene blue in the blood plasma to-be-measured sample. According to the method disclosed by the invention, the lowest detectable limit is 0.012mumol / L and the unexpected effect is obtained.

Description

technical field [0001] The invention relates to a method for detecting the content of methylene blue in blood plasma, belonging to the fields of clinical testing, drug analysis, biological analysis and analytical chemistry. Background technique [0002] In order to reduce the risk of clinical blood transfusion virus infection and enhance the safety of blood transfusion, methylene blue is generally used to inactivate the virus in the plasma, that is, the photochemical virus inactivation method, that is, by adding a certain amount of photosensitizer methylene blue to the plasma, A photochemical reaction occurs under light, thereby inactivating the virus in the plasma. In order to ensure the safety of plasma, it is necessary to ensure that there is a sufficient concentration of photosensitizer methylene blue (that is, a sufficiently high release amount) in the plasma before inactivation, and that there must be a sufficiently low concentration of photosensitizer methylene blue i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 谢洪平周群刚谢莲唐建华方敏徐军王明元
Owner 苏州市中心血站
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