Detection of free mannose and glucose in serum using high performance liquid chromatography
AssigneeThe Affiliated Hospital of Qingdao University
InventorsLijuan Zhang, Yong Liu
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Abstract
Provided is a method of detecting free mannose and glucose in serum using high performance liquid chromatography. Compared to existing technology, the pretreatment process of samples is simpler, the detection time is shortened, and the detection efficiency is greater. When detection of serum samples is carried out, mannose, rhamnose and glucose may be completely separated, and mannose and glucose will not affect each other during quantification, thereby ensuring the accuracy of detection results.
Claims10
A method of detecting free mannose and glucose in serum, using high performance liquid chromatography comprising:
- a) generating serum free monosaccharide comprising:
- (i) sequentially adding:
- ultrapure water, rhamnose solution, NaOH solution and 1-phenyl-3-methyl-5-pyrazolone solution into a serum sample and mixing to obtain a solution with a pH between 7-14, centrifuging the solution, and heating the solution for a predetermined time to obtain a sample A;
- (ii) cooling sample A to room temperature, adding HCl solution, mixing to obtain a solution with a pH between 1-7, centrifuging the solution, extracting with chloroform, wherein the supernatant becomes sample B;
- and
- (iii) centrifuging sample B at high-speed to obtain a sample C;
- and
- b) detecting sample C using high performance liquid chromatography;
- wherein the detection method is an external standard method, and rhamnose is used as internal reference monosaccharide.
Dependencies are inferred from the claim text.