A liquid chromatogram tandem mass spectrum method for measuring the content of D-pantothenic acid in urine

A technology of liquid chromatography and tandem mass spectrometry, applied in the field of D-pantothenic acid content, to achieve the effects of simple pretreatment, improved analysis throughput, and improved selectivity and accuracy

Active Publication Date: 2017-02-08
CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT +1
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Problems solved by technology

[0005] At present, the detection methods of D-pantothenic acid in urine mainly include high-performance liquid chromatography fluorescence detection (HPLC-FL) and liquid chromatography-mass spectrometry (LC-MS), such as Fukuwatari et al. established a method for the determination of D-pantothenic acid in urine with a quantitative limit of 32.5 µg / mL; Heudi et al. established an LC-MS method for the content of D-pantothenic acid in urine. The urine was simply centrifuged, diluted After detection and analysis, the detection limit of the method is 0.1 μg / mL, and LC-MS / MS has high sensitivity and high selectivity, but there is no method using LC-MS / MS to measure the content of D-pantothenic acid in urine. to report

Method used

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  • A liquid chromatogram tandem mass spectrum method for measuring the content of D-pantothenic acid in urine
  • A liquid chromatogram tandem mass spectrum method for measuring the content of D-pantothenic acid in urine

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example 1

[0030] 1. Instruments and reagents: Agilent 1200 liquid chromatography (Agilent, USA), equipped with G1329A autosampler, G1311A quaternary mixing pump and G1316A, column oven; API 4000 triple quadrupole tandem mass spectrometer (Applied Biosystems, USA) ), equipped with electrospray ion source (ESI) and analyst 1.5 software data processing system.

[0031] The standard D-pantothenic acid was purchased from Wuhan Yitai Technology Co., Ltd. Shanghai Branch. The purity was greater than 98%; both formic acid and acetonitrile were chromatographically pure, and the water used in the experiment was deionized water.

[0032] 2. Sample treatment: Thaw urine at room temperature, then pipette 100 µL of urine into a 10 mL volumetric flask, dilute to volume with 0.1% formic acid aqueous solution, mix well, then pipette about 1 mL into a 1.8 mL chromatographic bottle , to be analyzed by LC-MS / MS.

[0033] 3. Determination method: draw standard solution (concentrations of standard workin...

example 2

[0034] Example 2: According to the determination method described in Example 1, 8 urine samples were selected, and the content of D-pantothenic acid was measured as shown in Table 2.

[0035] Table 2 Content levels of D-pantothenic acid in samples (µg / mL)

[0036] serial number D-pantothenic acid serial number D-pantothenic acid 1 15.6 5 7.99 2 10.7 6 11.5 3 8.94 7 12.0 4 6.88 8 20.1

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Abstract

A liquid chromatogram tandem mass spectrum method for measuring the content of D-pantothenic acid in urine is disclosed. The method is characterized in that the method includes diluting a urine sample and then directly measuring by a liquid chromatogram-electrospray ionization-tandem mass spectrometer, an analyte is quantified by adopting an external standard process, and the content of the D-pantothenic acid in urine can be accurately measured. The method is a novel method for measuring the D-pantothenic acid in urine, the tandem mass spectrum increases selectivity and accuracy of the method, the limit of quantitation of the D-pantothenic acid is 1.54 ng / mL, the recovery rate of the method is 87.9-95.3%, and RSD is lower than 13.0%. Through selection and optimization of a chromatographic column and elution conditions, a chromatographic separating process is improved, and chromatographic analysis time is shortened and is 8 min.

Description

[0001] Technical field: [0002] The invention belongs to the technical field of physical and chemical testing of urine, in particular to a method for measuring the content of D-pantothenic acid in urine by using liquid chromatography-electrospray ionization-tandem mass spectrometer. [0003] Background technique: [0004] D-pantothenic acid is the precursor of coenzyme A (CoA). CoA is the biologically active form of its function. Pantothenic acid is required to participate in the production of energy from carbon-containing substances and the formation of many cell structures. Insufficient food intake or changes in physiological status may cause Causes pantothenic acid deficiency. The lack of pantothenic acid inhibits the reaction that requires CoA, mainly affects the metabolism of fatty acids, and inhibits the β-oxidation of fatty acids, especially the oxidation of fatty acids in peroxisomes is significantly inhibited, which can lead to hyperlipidemia, etc. damage. In additi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/02
CPCG01N30/02G01N30/88
Inventor 胡清源侯宏卫张小涛陈欢刘勇王安刘彤韩书磊付亚宁
Owner CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT
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