Quantify 15 bile acids in human plasma using Anyeep TQ9100.
This study used the Anyeep TQ9100 ultra-high-performance liquid chromatography-tandem mass spectrometry system from Anyeep to develop a method for simultaneously quantifying 15 bile acids in human plasma. The method demonstrated excellent linearity and met relevant regulatory requirements for limits of quantification. Using this method and equipment enables accurate detection and quantification of bile acids.
Bile acids are the primary functional components of bile and participate in the enterohepatic circulation., itsBile acid levels are a key indicator of liver and gallbladder function. Currently, bile acid testing is primarily used to screen for Intrahepatic Cholestasis of Pregnancy (ICP).
CurrentlyClinically, LC-MS/MS is the preferred method for analyzing and quantifying 15 bile acids due to its simplicity, sensitivity, and high specificity. Accordingly, this study employs the Anyeep TQ9100 liquid chromatography-tandem mass spectrometry system to measure the following bile acids in human serum: cholic acid (CA), glycocholic acid (GCA), taurocholic acid (TCA), deoxycholic acid (DCA), glycodeoxycholic acid (GDCA), taurodeoxycholic acid (TDCA), chenodeoxycholic acid (CDCA), glycchenodeoxycholic acid (GCDCA), taurochenodeoxycholic acid (TCDCA), ursodeoxycholic acid (UDCA), glycoursocholic acid (GUDCA), tauroursocholic acid (TUDCA), lithocholic acid (LCA), glycolithocholic acid (GLCA), and taurocholic acid (TLCA).
Instruments and Reagents
Anyeep TQ9100 Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry System
Acetonitrile (Mass Spectrometry Grade, Thermo Fisher Scientific, USA)
Water (Ultra-pure Water)
Formic Acid (Merck KGaA, Germany)
Liquid Chromatography Conditions
Mobile PhaseA: 0.1% formic acid, water
Mobile Phase B: Acetonitrile
Chromatography column：Agilent Poroshell 120 EC C18 column2.1mm×100mm 1.9 μm
Column Temperature：40 ℃
Flow rate: 0.25 mL/min
Injection volume：5 μL
Mass Spectrometry Conditions
Ionization Source： Electrospray Ionization Source (ESI)﹣）
Source Temperature： 550 ℃
Spray Voltage： 2500 V
air curtain： 10 psi
Mist gas： 50 psi
Auxiliary Gas： 55 psi
Crash： 8
Scan Mode：MRM
Sample Processing
Retrieve test sample 100 μL, add 20 μL internal standard, 380 μL acetonitrile, vortex and centrifuge (4 °C, 15000) rpm, 5 min); collect supernatant 400 μL in 96-well plate; evaporate to dryness with nitrogen at 40 °C; add 100 Reconstitute in μL of solution (50% acetonitrile) on a microplate shaker for 1200. rpm/min shaking 5 min, 4000 RPM centrifugation at 5 min, 5 μL injection.
Summary
This method utilizes the TQ 9100 triple quadrupole mass spectrometry system, independently developed and manufactured by Anyipu, to achievePlasmaSimultaneous detection of 15 bile acids with excellent linearity and high accuracy.High specificityEach sample analysis takes only 6 minutes, offering high throughput.Suitable for clinical diagnostic analysis。