Detection kit for detecting fat-soluble vitamins in serum by high performance liquid chromatography-tandem mass spectrometry and detection method thereof

Abstract

The invention provides a detection kit for detecting fat-soluble vitamins in serum by high performance liquid chromatography-tandem mass spectrometry and a detection method thereof. Methanol, acetonitrile, isopropanol and an internal standard substance working solution are mixed to prepare an internal standard solution, and ammonium acetate is added into the internal standard solution, so that sample pretreatment is simply and efficiently completed; the recovery rate of vitamins A, E, K1 and K2 in the serum sample can be obviously improved without any additional sample enrichment operation, the pretreated sample is more stable, and the detection sensitivity of fat-soluble vitamins in the serum is greatly improved; and the prepared internal standard solution is very stable, can be stored for a long time and can be taken at any time, so that the detection process is simpler, more convenient and more efficient, the detection cost is lower, and the detection result is more accurate and stable. Meanwhile, the invention further provides application of ammonium acetate to preparation of an internal standard solution.

Description

technical field [0001] The invention relates to the technical field of chemical analysis, in particular to a detection kit and a detection method for detecting fat-soluble vitamins in serum by high performance liquid chromatography tandem mass spectrometry. Background technique [0002] Vitamin A (VA), also known as retinol, has the functions of maintaining normal vision, maintaining epithelial tissue, and promoting normal growth and development of bones. Insufficient VA in the human body can lead to nutritional deficiencies such as dry eye disease, night blindness, and hyperkeratosis of the skin. 25-Hydroxyvitamin D is an important factor that regulates the metabolism of calcium and phosphorus in the human body, coordinates the mobilization or deposition of bone calcium, the absorption or excretion of urinary calcium, maintains the stability of blood calcium levels in the body, and plays an important role in bone development and shaping, muscle nerve conduction effects and ...

AI Technical Summary

Problems solved by technology

[0003] At present, there are many reports on the determination methods of fat-soluble vitamins in serum at home and abroad. Common methods include: chromatography, spectrophotometry, gas chromatography, and liquid chromatography. Among many methods, liquid chromatography or liquid chromatography Tandem mass spectrometry is the preferred method for the determination of fat-soluble vitamins. Previous methods have problems such as complex operation process, many interference factors, poor specificity, long analysis time, low sensitivity, poor qualitative and quantitative accuracy, and poor reproducibility. The reason is that the sample pretreatment process is cumbersome, the recovery rate is not high, and the stability is poor.

[0004] Existing fat-soluble vitamin sample pretreatment methods mainly adopt liquid-liquid extraction (such as CN106504947A) and solid-liquid extraction method (such as CN110763788A), need multiple steps such as dilution, multiple extraction, rotary evaporation, nitrogen drying, purification, etc. , the process is cumbersome and requires a lot of manual operations, and it is difficult to achieve high-throughput and automation for liquid-liquid extraction. Although solid-liquid extraction can achieve high-throughput, the sample extraction process is mainly completed by SLE plates, and the cost of consumables is high.

[0005] At present, there are also many related patent applications for sample pretreatment using protein precipitation method for the determination of fat-soluble vitamin samples in serum and plasma, such as CN111999397A, CN110487943B, etc., but there is generally a problem of low recovery rate resulting in low sensitivity, and some even After protein precipitation, further extraction is required for subsequent detection and analysis, which makes clinical detection require more manual operations, which is not conducive to clinical promotion and use

Images