Method for detecting bile acid in excrement by UPLC-MS/MS
A bile acid and feces technology, which is applied in the field of bile acid detection in feces, can solve the problems of insufficient detection range of bile acid, and achieve the effect of absolute quantification, strong specificity, and high sensitivity
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Embodiment 1
[0046] Internal standard working solution preparation:
[0047] Firstly, chenodeoxycholic acid-D4, cholic acid-D4, ursodeoxycholic acid-D4, glycochenodeoxycholic acid-D4, glycocholic acid-D4 and lithocholic acid- D4 was prepared into 1mg / mL internal standard mother solution respectively, and then the mother solution was diluted according to the concentration shown in Table 1 to obtain the internal standard stock solution, and the internal standard stock solution was processed in the next step according to the requirements in Table 1 to obtain the internal standard work liquid.
[0048] Table 1
[0049]
[0050] Chromatographic conditions:
[0051] Chromatographic column: Agela Venusil MP C18 2.1mm×100mm×2.5μm;
[0052] Mobile phase: A phase: 0.1% formic acid aqueous solution; B phase: acetonitrile;
[0053] Column temperature: 50°C;
[0054] Injection volume: 2μL;
[0055] The chromatographic gradient is shown in Table 2.
[0056] Table 2
[0057] time min...
Embodiment 2
[0082] Compared with Example 1, the difference is that the chromatographic gradient is as shown in Table 4.
[0083] Table 4
[0084] time min Flow rateml / min A% B% 0.0 0.5 80 20 0.5 0.5 80 20 2 0.5 65 35 10 0.5 50 50 16 0.5 0 100 16.1 0.5 80 20 19 0.5 80 20
[0085] According to the chromatographic gradient of Example 2, the mixed standard sample is separated, and the separation effect is worse than that of Example 1. The specific performance is that the peaks are unevenly distributed, and there are miscellaneous peaks at the end of the gradient. For details, see Figure 5 The total ion chromatogram is shown.
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