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Kit for simultaneously quantifying and detecting niacin, nicotinamide and pantothenic acid

A quantitative detection and nicotinamide technology, applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems that the vitamins to be tested cannot meet the detection limit and cannot be quantified, and achieve high accuracy, convenient storage and transportation, good precision effect

Inactive Publication Date: 2016-09-21
首都儿科研究所 +1
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Problems solved by technology

The combination of DBS and LC-MS / MS technology has become an important method in the quantitative analysis of small molecules, but in the field of vitamin quantification, there are no reports at home and abroad on the application of DBS combined with LC-MS / MS technology for the quantification of B vitamins
In addition to the sample pretreatment and separation problems that need to be solved in liquid whole blood, due to the small amount of whole blood collected (50 μ), it is also prone to the situation that the vitamins to be tested cannot meet the detection limit and cannot be quantified.

Method used

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  • Kit for simultaneously quantifying and detecting niacin, nicotinamide and pantothenic acid
  • Kit for simultaneously quantifying and detecting niacin, nicotinamide and pantothenic acid
  • Kit for simultaneously quantifying and detecting niacin, nicotinamide and pantothenic acid

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Embodiment 1

[0036] Select 15 dried whole blood spots of children under 10 years old, take a blood spot, which is equivalent to 50 μL of whole blood, add 50 μL of isotope internal standard solution, 450 μL of 200 g / L trichloroacetic acid aqueous solution, vortex and soak the liquid surface Dried blood slices were extracted by ultrasound for 30 minutes in the dark, vortexed for 5 minutes, centrifuged at 13,000 rpm for 5 minutes, and 40 μL was taken for LC-MS / MS analysis. After calculation by the standard curve of the same batch, the contents of the three B vitamins in the whole blood were quantitatively obtained (see Table 7).

[0037] Figure 1-3 For the standard working curves of niacin, nicotinamide, and pantothenic acid, the abscissa is the ratio of the concentration of the vitamin to the corresponding isotope internal standard, and the ordinate is the ratio of the response of the vitamin to the corresponding isotope internal standard. The weighting method is 1 / X2.

[0038] Figure 4-...

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Abstract

The invention provides a kit for quantitatively detecting niacin, nicotinamide and pantothenic acid in peripheral whole blood at the same time, which mainly consists of dried blood slices (Whatman protein saver 903), capillary blood collection tubes, nicotinic acid and nicotinamide and pantothenic acid vitamin standard substance and its isotope internal standard, trichloroacetic acid, and also includes dry blood spot preparation, dry blood spot pretreatment and detection steps. The nicotinic acid, nicotinamide, and pantothenic acid dry blood spot quantitative kit provided by the present invention combines dry blood spot sampling technology with high performance liquid chromatography-tandem mass spectrometry technology for the quantification of B vitamins, a minimally invasive, minimal blood collection at the same time Realize the evaluation of the nutritional status of multiple B vitamins, and realize the quantitative work of simultaneous detection of multiple vitamins under low sample volume conditions that cannot be accomplished by metabolic enzyme-dependent methods and immunoassay methods.

Description

technical field [0001] The invention belongs to the field of vitamin quantitative detection, in particular to a kit for simultaneous quantitative detection of nicotinic acid, nicotinamide and pantothenic acid in whole blood dried blood spots. Background technique [0002] Pantothenic acid (VB5), niacin (VB3), and nicotinamide are B vitamins with small molecular weight, and their content in the human body is very low, but they are crucial to the maintenance of normal physiological functions of the body. Nicotinic acid (VB3) and nicotinamide participate in the central electron transfer and hydrogen transfer of cell metabolism through the formation of nicotinamide adenine dinucleotide (NAD) and nicotinamide adenine dinucleotide phosphate (NADP), and participate in at least 200 Carbohydrate, fatty acid and amino acid metabolic reactions. Pantothenic acid (VB5) is an essential component of coenzyme A (CoA), and acetyl CoA formed by CoA and acetic acid is a key factor in the meta...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/06G01N30/89G01N33/82
CPCG01N30/88G01N30/06G01N30/89G01N33/82G01N2030/062G01N2030/065G01N2030/8822
Inventor 张霆刘鸿君吴建新霍军生张敏邵立军杜克贺王国才
Owner 首都儿科研究所
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