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- W4381125616 abstract "<h3>Introduction</h3> 7α-Hydroxy-4-cholesten-3-one (C4) is the common intermediary of both primary bile acids (BAs). Its concentration in serum correlates with the rate of hepatic BA production. C4 was recommended, alongside <sup>75</sup>SeHCAT, for the investigation of Bile Acid Diarrhoea (BAD) in patients with chronic diarrhoea by the British Society of Gastroenterology 2018 guidelines. This project aimed to develop and validate an assay to quantitate C4 in serum for the investigation of BAD and assess if C4 is stable enough in blood for testing of patients in primary care. <h3>Methods</h3> The method was developed using a Waters TQS Mass Spectrometer. Method accuracy was underpinned by calibrating to quantitative nuclear magnetic resonance analysis. C4 was analysed in a high-throughput 96-well plate format using deuterated C4 as an internal standard. Simple liquid-liquid extraction was performed and full validation criteria assessed. To assess C4 stability, samples (n=8) were collected from healthy volunteers (n=12). Samples were incubated at 20°C for up to 72 hours and retrieved, centrifuged and frozen for storage at different time points prior to C4 analysis. <h3>Results</h3> The C4 method demonstrated excellent analytical performance and passed all validation criteria. The method was found to be accurate, precise, free from matrix effects and was not susceptible to assay interference. C4 5—1000 nmol/L could be reliably quantitated. In unseparated blood, C4 concentration was found to decrease over time. After 2, 4, 8, 12, 24, 48 and 72 hours mean C4 changes were -1.2% (95% CIs: -2.4 — 0.0%), -1.1% (-2.0 — -0.2%), -2.3% (-3.3 — -1.3%), -2.6% (-4.6 — -0.6%), -4.4% (-6.4 — - 2.4%), -8.9% (-10.7 — - 7.2%), and -12.6% (-14.3 — - 10.8%), respectively <h3>Conclusions</h3> We report a robust and accurate method of analysing C4 in serum, offering a convenient option alongside <sup>75</sup>SeHCAT for BAD investigation. C4 was found to degrade in unseparated blood over time, however after 48 hours the mean change was < -9% from baseline. This enables collection of samples from both primary and secondary care, which should improve the patient pathway, but is unsuitable for home collection." @default.
- W4381125616 created "2023-06-19" @default.
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- W4381125616 date "2023-06-01" @default.
- W4381125616 modified "2023-09-27" @default.
- W4381125616 title "P164 Bile acid malabsorption: the development of a novel serum 7α-hydroxy-4-cholesten-3-one (C4) method to aid diagnosis" @default.
- W4381125616 doi "https://doi.org/10.1136/gutjnl-2023-bsg.235" @default.
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