Method for quantitatively analyzing retinol and precursor thereof on dry blood spot card with tiny amount of blood

A technology for dry blood filter paper and retinol, which is applied in the field of quantitative analysis of retinol and its precursors on trace blood dry blood filter paper, can solve the problem of lack of extraction of retinol, and achieve good linear relationship and high precision , the effect of high sensitivity

Active Publication Date: 2015-09-16
CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, there is still a lack of international and domestic methods f

Method used

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  • Method for quantitatively analyzing retinol and precursor thereof on dry blood spot card with tiny amount of blood
  • Method for quantitatively analyzing retinol and precursor thereof on dry blood spot card with tiny amount of blood
  • Method for quantitatively analyzing retinol and precursor thereof on dry blood spot card with tiny amount of blood

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[0048] Example 1

[0049] (1) Materials and methods

[0050]

[0051] main reagent

[0052] Reagent preparation: retinol, retinyl acetate, retinyl palmitoyl ester standard dissolved in 0.1% (w / v) in the methanol solution, the β-carotene standard substance is dissolved in the dichloromethane containing BHT 0.1% (w / v), and the preparation concentration is respectively 5mg / ml, 2mg / ml, 2mg / ml, 5mg / ml The stock solution in ml is diluted 1000 times with methanol containing 0.1% (w / v) BHT to be the working solution. The working solution is freshly prepared every week, protected from light, and stored at -20°C.

[0053] Stable isotope-labeled 19, 19, 19, 20, 20, 20-d6-retinyl acetate as internal standard, dissolved in methanol containing BHT 0.1% (w / v), 0.1 μmol / L as working solution concentration.

[0054] 1. Establishment of HPLC-MS / MS method for the analysis of retinol and its precursors in dried blood filter paper

[0055] 1.1 Preparation of dried blood filter pa...

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Abstract

The invention provides a method for detecting retinol and a precursor thereof. The method comprises the following steps: 1, extracting retinol and/or the precursor thereof from a sample; 2, separating retinol and/or the precursor thereof through HPLC; and 3, carrying out MS/MS quantitative detection, wherein the sample is a dry blood spot card sample, and an extractant adopted in the invention is trichloromethane. A filter paper card is adopted as a carrier to adsorb a tiny amount of blood in order to make the dry blood stop card, so the difficulties of much blood induced by venous blood collection, bad population compliance, death induced by animal model blood drawing, and inconvenient long-distance conveying of samples in previous serum collection and detection process are overcome. The method has the advantages of small matrix effect, good linear relation of analysis in the concentration range of a standard curve, low detection limit, high sensitivity, good accuracy and high precision, is related with the result of detection of retinol in serum through HPLC, allows the contents of the retinol and the precursor thereof in the sample to be simultaneously and synchronously analyzed, and provides a method for comprehensive assessment of the in vivo retinol level.

Description

technical field [0001] The invention relates to an analysis and detection method for retinol and its precursor in trace blood dried blood filter paper sheets. Background technique [0002] Retinol in the blood needs to be ingested from animal and plant foods, and its precursors: β-carotene, retinyl acetate and retinyl palmitoyl ester are transformed and formed in the intestinal mucosa and liver cells, and co-exist in the blood. Only by combining information on the content of retinol and its precursors can we fully measure the level of retinol in the body. [0003] Currently, fluorescence, ultraviolet spectrophotometry and high performance liquid chromatography (High performance liquid chromatography, HPLC) are commonly used to quantitatively analyze retinol in serum or plasma. The extraction steps of the first two methods are cumbersome, and the results are greatly affected by impurities; while HPLC The method can separate complex components in serum, and its detection sen...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
Inventor 李廷玉苗静琨邹琳余加林杨亭
Owner CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV
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