A detection kit for detecting fat-soluble vitamins in serum by high performance liquid chromatography tandem mass spectrometry and its detection method
A technology of high performance liquid chromatography and fat-soluble vitamins, applied in the field of chemical analysis, can solve problems such as poor stability, high cost of consumables, difficulty in achieving high-throughput and automation of liquid-liquid extraction, etc., to ensure precision and accuracy, improve Accuracy and reliability, to ensure the effect of sample separation effect
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Embodiment 1
[0062] Embodiment 1: sample preparation, pretreatment, detection, analysis
[0063] 1. Sample preparation
[0064] 1. Preparation of standard curve and quality control samples
[0065] Prepare vitamin A, 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, vitamin E, vitamin K1, and vitamin K2 standard products into a mixed solution, which is used as the stock solution of the standard working solution and the quality control working solution, and then mixed with the negative blank Serum and negative serum were mixed at a volume ratio of 1:49 to prepare standard curve and quality control samples.
[0066] The six fat-soluble vitamins have 10 serial concentrations (S1-S10) in the standard product, as shown in Table 1:
[0067] Table 1. Concentrations of 10 series of 6 fat-soluble vitamins in standard products (S1~S10)
[0068]
[0069]
[0070] The 6 kinds of fat-soluble vitamins have three series concentrations of low (L), medium (M) and high (H) in quality control products, as...
Embodiment 2
[0115] Example 2: Comparison of long-term storage stability evaluation results of different internal standard solution systems
[0116] In this embodiment, according to the internal standard solution preparation method provided in Example 1, an internal standard solution containing 6 kinds of fat-soluble vitamin internal standards was prepared, and different internal standard solution system components as shown in Table 7 were used to carry out stability tests respectively. The stability test refers to putting it in a 48°C incubator for six months to observe whether the internal standard solution has any abnormal phenomena such as turbidity and deterioration, and compare whether the internal standard solution that has been tested for stability is different from the newly prepared internal standard solution. The detection method provided in Example 1 detects whether the concentration deviation of the internal standard component occurs, and whether the recovery rate of the target...
Embodiment 3
[0121] Embodiment 3: Comparison of detection results using different internal standard solution systems
[0122] According to the internal standard solution preparation method provided in Example 1, the present embodiment prepares internal standard solutions containing 5 kinds of fat-soluble vitamin internal standards, adopts different internal standard solution system components as shown in Table 8, and provides according to Example 1 Step method After sample preparation and pretreatment, take the lowest concentration point (S1) of the standard curve and enter the liquid chromatography-mass spectrometry system for analysis. The peak areas of the six fat-soluble vitamins in the S1 sample are as shown in Table 8:
[0123] Table 8. Comparison of detection results using different internal standard solution systems
[0124]
[0125] As can be seen from Table 8, when using the internal standard solution system 50mM ammonium acetate (75% methanol+15% acetonitrile+10% isopropanol)...
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Abstract
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