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Method for reducing non-specific adsorption of fat-soluble vitamin 96-hole plate

A fat-soluble vitamin, non-specific technology, applied in the non-specific adsorption during sample pretreatment, to reduce the field of high-performance liquid phase detection of fat-soluble vitamins, can solve the problems affecting the accuracy, low recovery rate of fat-soluble vitamins, etc., to improve the accuracy The effect of reducing non-specific adsorption problems and improving efficiency

Inactive Publication Date: 2020-04-10
JINAN YING SHENG BIOTECH
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0006] The above pretreatment process involves the elution process, as well as the subsequent nitrogen blowing and reconstitution process. The above process requires a matching 96-well collection plate, so the sample remains in the 96-well collection plate for a long time, and in several experiments It was found that the recovery rate of fat-soluble vitamins was low. The inventor analyzed that the reason may be that the 96-well collection plate is made of plastic, which is easy to cause non-specific combination with fat-soluble vitamins, especially vitamin D and vitamin K. The defects of the above equipment have seriously affected the detection. the accuracy of

Method used

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  • Method for reducing non-specific adsorption of fat-soluble vitamin 96-hole plate
  • Method for reducing non-specific adsorption of fat-soluble vitamin 96-hole plate
  • Method for reducing non-specific adsorption of fat-soluble vitamin 96-hole plate

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Embodiment 1

[0045] In this example, a pre-treatment method for the detection of fat-soluble vitamin samples is provided, which is a treatment method for reducing non-specific adsorption on a 96-well collection plate.

[0046] 1) First, the 96-well collection plate is autoclaved, and the conditions for autoclaving are explored;

[0047] 2) Put the serum sample and the internal standard working solution in a 96-well extraction plate, and mix by pipetting;

[0048] 3) Transfer all the liquid in 2) to a 96-well filter plate (equipped with an autoclaved 96-well collection plate), press the liquid into the small column with a positive pressure device, and let it stand for 5-10 minutes;

[0049] 4) Add the extract, and collect the eluate in a 96-well collection plate with a positive pressure device. This step is repeated once.

[0050] 5) Place the 96-well collection plate at room temperature and dry it with nitrogen gas (flow rate of 2 mL / min, about 5 min).

[0051] 6) Reconstitute and test ...

Embodiment 2

[0053] In this embodiment, a method for determining the content of fat-soluble vitamins (vitamin A, vitamin E, vitamin K, 25-OH VD2, 25-OH VD3) in serum samples is provided.

[0054] 1.1 Preparation of samples to be tested

[0055] Take 300 μL of serum to a 96-well extraction plate, add 100 μL of internal standard working solution, and mix by pipetting; transfer all the above liquid to a 96-well filter plate (equipped with a 96-well collection plate), and press the liquid into the small column with a positive pressure device Inside, let stand for 5-10min. Add 750 μL of extraction solution, and collect the eluate in a 96-well collection plate with a positive pressure device. This step is repeated once. Place at room temperature, blow dry with nitrogen, redissolve in the reconstituted solution, shake and mix for 2 minutes, cover with aluminum foil, and serve as the sample to be tested.

[0056] The above extract is n-hexane.

[0057] 1.2 Preparation of standard working solut...

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Abstract

The invention, which belongs to the technical field of fat-soluble vitamin detection pretreatment, particularly relates to a method for reducing non-specific adsorption of a fat-soluble vitamin 96-hole plate. Commercial 96-hole plates have important application in the sample pretreatment process. The inventor finds that the material of the 96-hole collecting plate has high non-specific adsorptionon fat-soluble vitamins in the research process, and the subsequent nitrogen blowing step can influence the detection of the fat-soluble vitamins. The invention provides a method for effectively reducing non-specific adsorption of a 96-hole plate to fat-soluble vitamins; the method comprises the step of pretreating a 96-hole collecting plate by adopting high-pressure sterilization; and the research proves that the treatment method can effectively reduce the specific adsorption of the 96-hole collecting plate to the fat-soluble vitamins and improve the recovery rate.

Description

technical field [0001] The invention belongs to the technical field of fat-soluble vitamin detection pretreatment, and in particular relates to a method for reducing non-specific adsorption of fat-soluble vitamins before high-performance liquid phase detection and during sample pretreatment. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Fat-soluble vitamins (including VA, VE, VK and the main metabolites 25-OH VD2 and 25-OH VD3 of VD) are low-molecular organic compounds that are indispensable for maintaining normal substance metabolism and certain special physiological functions in the human body, and can regulate Various physiological functions of the human body, enhanc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88
Inventor 冯振景叶松弭兆元
Owner JINAN YING SHENG BIOTECH
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