Method for detecting concentrations of eight flavonoid compounds in human urine by UPLC-MS/MS (ultra performance liquid chromatography-mass spectrometry/mass spectrometry) method

Abstract

The invention discloses a method for detecting the concentration of eight flavonoid compounds in human urine by a UPLC-MS/MS method, namely the detection of quercetin, hesperetin, myricetin, naringenin, phloretin, isorhamnetin, apigenin and kaempferol. Firstly, an object to be detected is separated from a urine matrix by utilizing ultra-high performance liquid chromatography, a standard curve is established by taking the concentration of the standard substance as an X axis and taking the area of a quantitative ion peak in the standard substance as a Y axis by utilizing a mass spectrometry quantitative method to calculate the concentrations of the eight flavonoid compounds. The method comprises the following steps: concentrating and redissolving the sample by using a freeze dryer, analyzing the sample by using an ultra-high performance liquid chromatography-tandem mass spectrometer (UPLC-MS), pre-freezing moisture and an organic solvent in the sample by using the freeze dryer, directly sublimating in a vacuum state, and capturing evaporated gas by using a cold trap, so that the evaporation process is quickly carried out, using 24-hour freeze drying for replacing nitrogen blowing, so that the sample pretreatment step can be simplified, the detection efficiency is improved, and the method is suitable for large-sample detection of flavonoid compounds in human urine.

Description

technical field [0001] The invention relates to the field of biological monitoring, in particular to a method for detecting the concentrations of eight flavonoid compounds in human urine by UPLC-MS / MS. Background technique [0002] In recent years, the health-promoting benefits of fruits and vegetables have attracted increasing attention from scholars at home and abroad. A large number of epidemiological survey data show that the intake of fruits and vegetables can effectively regulate blood sugar and blood lipid metabolism, and reduce the risk of chronic diseases such as type 2 diabetes, metabolic syndrome, and stroke. At present, the commonly used evaluation tool for fruit and vegetable intake is the meal frequency questionnaire (FFQ). This semi-quantitative questionnaire is not accurate due to the inaccurate self-reported intake and it is difficult to control recall bias. . A large number of studies have shown that flavonoids in urine can be used as biomarkers of fruit ...

AI Technical Summary

Problems solved by technology

[0004] First, there is a risk of blowing the substance in the sample into the laboratory when nitrogen blowing is performed with a nitrogen blowing device, and it must be operated in a fume hood;

[0005] Second, nitrogen blowing flammable solvents has an explosion hazard, so samples containing flammable solvents cannot be concentrated by nitrogen blowing;

[0006] Third, when concentrating toxic solvents, even if the entire system is placed in a fume hood, the operator still has the risk of poisoning;

[0007] Fourth, the sample temperature is high during nitrogen blowing, which is not suitable for temperature-sensitive samples;

[0008] Fifth, the number of holes and gas needles of the nitrogen blowing instrument is limited. The common nitrogen blowing instruments are mainly 12 holes, 24 holes and 36 holes, and the sample processing volume is small, so it is impossible to process samples in large quantities at one time;

[0009] Sixth, the flow of nitrogen gas needs to be kept constant and uniform during the nitrogen blowing process, so the operator needs to constantly adjust the gas flow according to the sample liquid level to monitor the whole process; if the gas flow is not well controlled, it is easy to cause sample splashing and loss of samples , it is also easy to cause mutual contamination of samples;

[0010] Seventh, nitrogen blowing needs to consume nitrogen, increasing additional costs

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