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Method for researching reaction behavior of PQQ (Pyrroloquinoline quinone), and Lys (Lysine) and Arg (Arginine)

A technology of reaction and behavior, applied in the field of biological analysis, can solve problems such as incomplete understanding of the mechanism of action

Active Publication Date: 2018-12-25
JIANGSU INST OF NUCLEAR MEDICINE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, its mechanism of action is not fully understood

Method used

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  • Method for researching reaction behavior of PQQ (Pyrroloquinoline quinone), and Lys (Lysine) and Arg (Arginine)
  • Method for researching reaction behavior of PQQ (Pyrroloquinoline quinone), and Lys (Lysine) and Arg (Arginine)
  • Method for researching reaction behavior of PQQ (Pyrroloquinoline quinone), and Lys (Lysine) and Arg (Arginine)

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Experimental program
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Effect test

Embodiment 1

[0037] 1. UPLC chromatographic separation conditions

[0038] UPLC analysis conditions, column: ACQUiTY UPLC@ BEH C18 column (2.1×100mm, 1.7μm), mobile phase A: H 2 O+0.1% formic acid, B: Methanol; 2 / 98 B / A was eluted for 5 minutes, and gradient eluted to 60 / 40 B / A in 20 minutes. PDA detection. After adding 0.1% formic acid to the mobile phase and gradient elution, PQQ can be separated from the reaction product better.

[0039] 2. Standard curve, precision and detection limit determination

[0040] Accurately weigh a certain amount of PQQ, prepare standard solutions with concentrations of 0.2, 0.4, 0.6, 0.8, and 1.0 mM, inject samples separately, plot the obtained integral area against the sample concentration, and perform linear regression, and the regression equation is y =6E +06x-7834.9, R 2 =0.9998; linear range 0.2~1.0mM; such as figure 1 shown.

[0041] Under the above-mentioned chromatographic conditions, 6 consecutive injections were made on the same day, and th...

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Abstract

The invention discloses a method for researching reaction behavior of PQQ, and Lys and Arg. The method is a first part of the research on the inhibition effect of PQQ and AGEs (advanced glycation endproducts), and belongs to the field of bioanalysis technology. The Lys and the Arg are main free amino acids of DNA involved in the glucose reaction. In order to clarify the mechanism of action, the method researches the reaction behavior of PQQ, and Lys and Arg, optimizes UPLC (ultra performance liquid chromatograph) chromatographic separation conditions, avoids the damage to the chromatographiccolumn by using a high concentration salt solution, and also separates the enantiomers. The method comprises the steps of: determining standard curve, accuracy and detection limit of the PQQ concentration; measuring free PQQ concentration during the reaction of the PQQ with Arg and Lys; identifying structure of reaction product of the PQQ and Lys or Arg by UPLC-MS. According to measurement mass spectrum result, new compound PQQ-Lys, pqq-Arg is found, and it also find that one molecular of PQQ can simultaneously react with one molecule of Lys and one molecular of Argto obtain a new compound. The PQQ inhibits the formation of AGEs by reacting with Lys and Arg to provide experimental basis for the possible mechanisms of PQQ-inhibition AGEs formation.

Description

technical field [0001] The method for studying the reaction behavior of PQQ (pyrroloquinoline quinone) with Lys (lysine) and Arg (arginine) is the first part of the research on the inhibitory effect of PQQ and advanced glycation end products AGEs, which belongs to the field of bioanalysis technology . Background technique [0002] Advanced glycation end products (AGEs) are chronic non-enzymatic glycation products of proteins. AGEs are ubiquitous in normal people, and the level of plasma AGEs will also increase with age. It is an important biomarker of body aging, and the level of AGEs in diabetic patients and the elderly generally increases. It has been reported that the formation of AGEs may occur during early plaque formation in Alzheimer's disease (AD). AGEs bind to RAGE (AGEs receptor) on the cell surface and can cause oxidative stress, triggering an inflammatory cascade, atherosclerosis, and even thrombosis. The accumulation of AGEs in vivo causes multiple complicati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 蔡刚明周杏琴钦晓峰徐希杰顾晓波张荣军俞惠新包建东
Owner JIANGSU INST OF NUCLEAR MEDICINE
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