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Novel method for measuring nucleotide content in infant formula milk powder

An infant formula and nucleotide technology, which is applied to measuring devices, instruments, and material analysis through electromagnetic means, to achieve the effects of improving qualitative ability, reducing labor rate, and improving inspection speed

Inactive Publication Date: 2012-01-25
WISSUN INT NUTRITION GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, existing methods have significant limitations

Method used

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  • Novel method for measuring nucleotide content in infant formula milk powder
  • Novel method for measuring nucleotide content in infant formula milk powder
  • Novel method for measuring nucleotide content in infant formula milk powder

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Experimental program
Comparison scheme
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Embodiment Construction

[0016] The present invention is described in detail below in conjunction with embodiment

[0017] 1. Experimental part

[0018] 1.1 Reagents

[0019] CMP, purity ≥ 98%, UMP, purity ≥ 99%, AMP, purity ≥ 99.0%, GMP, purity ≥ 99%, IMP, purity ≥ 99% were analytically pure (SIGMA). Formic acid, ammonium formate, acetic acid, ammonia water and ammonium acetate are of superior grade. The experimental water was 18.2 MΩ·cm ultrapure water.

[0020] equipment

[0021] Liquid chromatography tandem quadrupole mass spectrometer (Agilent1200RRLC, Agilent6410BQQQ, Masshanter software); Milli-Q water purification system (Millipore); vortex mixer; ultrasonic cleaner; high-speed centrifuge; PHS-3E laboratory pH count.

[0022] Standard solution preparation

[0023] Accurately weigh CMP, AMP, UMP, GMP, IMP, and ultrapure water to prepare a 1 mg / mL standard stock solution. Protect from light and store at low temperature.

[0024] Blank matrix solution preparation

[0025] Tak...

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Abstract

The invention discloses a novel method for measuring the nucleotide content in infant formula milk powder. The method comprises the following steps of: dissolving an infant formula milk powder sample in water; adjusting the pH to 4.6 with a formic acid solution; precipitating proteins in the sample; ultrasonically extracting, centrifuging and filtering; separating five types of nucleotides, i.e. CMP (Cytidine), AMP (Adenosine), UMP (Uridine), GMP (Guanosine) and IMP (Inosine) in a filtrate with a reversed phase chromatographic column; performing tandem mass spectrum; scanning in a multiple reaction monitoring (MRM) scanning mode; and quantifying with a substrate by using an external reference method. In the invention, five types of nucleotides in the infant formula milk powder are measured with an LC-MS / MS (Liquid Chromatogram-Mass Spectrometry / Mass Spectrometry) method, so that the using quantity of the sample is extremely small in an ESI (Electrospray Ionization) mode, an ion source is prevented from being polluted easily, and high sensitivity and low detection limit are realized. In particular, the MRM method is adopted, interference of non-nucleotide molecular ions can be well eliminated; and meanwhile, the method has the advantages of simple sample pretreatment and wide application range.

Description

technical field [0001] The present invention relates to a new method for determining nucleotide content in formula milk powder for infants and young children, in particular to a method for determining cytosine nucleotide and uracil nucleotide in formula milk powder for infants and young children by using liquid chromatography-tandem mass spectrometry and multi-reaction ion monitoring The invention discloses a detection method for five kinds of nucleotides including adenine nucleotide, guanine nucleotide and inosine nucleotide, belonging to the technical field of chemical composition detection. Background technique [0002] Nucleotides are composed of nitrogen-containing bases, ribose or deoxyribose sugars, and phosphates linked by three molecules. Bases and sugars are linked to form nucleosides by glycosidic linkages, and nucleosides and phosphates are linked to form nucleotides by ester linkages. Its main physiological function is to widely participate in the energy metabo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/60G01N27/62
Inventor 陈光耀林玉宙
Owner WISSUN INT NUTRITION GRP
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