LC–MS/MS-based quantification of cholesterol and related metabolites in dried blood for the screening of inborn errors of sterol metabolism

Becker, Stefan; Röhnike, S.; Empting, Susann; Haas, Dorothea; Mohnike, Klaus; Beblo, Skadi; Mütze, Ulrike; Husain, Ralf A.; Thiery, Joachim; Ceglarek, Uta

doi:10.1007/s00216-015-8731-1

Cited by 24 publications

(10 citation statements)

References 19 publications

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“…Sterols were determined based on the previously published liquid chromatography–mass spectrometry (LC-MS/MS) method [23]. In brief, 10 μl of calibrator, quality control, or sample (serum or plasma) were mixed with 490 μl of internal standard solution (100 μg/L d7-cholesterol in methanol/isopropanol, 1/1, v/v) in polypropylene tubes.…”

Section: Methodsmentioning

confidence: 99%

Low PCSK9 levels are correlated with mortality in patients with end-stage liver disease

et al. 2017

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IntroductionProprotein convertase subtilisin/kexin type 9 (PCSK9) plays a key role in the cholesterol metabolism and is synthesized by the liver. It interacts with the LDL-receptor to promote its degradation. The model of end-stage liver disease (MELD) score is a well-established tool to estimate the risk of mortality in patients with end-stage chronic liver disease. The study aims to assess the associations between PCSK9, hypocholesterinemia, liver synthesis, cholestasis, MELD score and mortality in patients with end-stage liver disease.MethodsSerum samples were obtained from 74 patients with severe liver disease. The study participants were aged between 23 and 70 y (mean: 55.8 y; 47 males [63.5%], 27 females [36.5%]). Samples were selected from those with a wide range of MELD scores (7 to 40).Patients that underwent liver transplantation (17 / 74) were censored at the time of transplantation for mortality analysis.ResultsPCSK9 values ranged from 31.47 ng/mL to 261.64 ng/mL. The median value was 106.39 ng/ml. PCSK9 was negatively correlated with markers of liver function and cholestasis (INR, bilirubin). Over a 90-d follow-up, 15 of 57 (26,3%) patients died within the 90-d follow-up without receiving liver transplantation. Thirteen of 31 (42%) patients with PCSK9 levels below the median died compared to 2/26 (8%) patients with higher PCSK9 levels (p = 0.006). In this cohort, there were no significant correlations between PCSK9, cholesterol, its precursors and several phytosterols.ConclusionsLow PCSK9 serum concentrations were associated with higher mortality in patients with end-stage liver disease. The mean PCSK9 levels in the study population were much lower than those found in normal or healthy populations. Further studies are required to acquire a more detailed understanding of the role of PCSK9 in liver-related mortality.

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Section: Methodsmentioning

confidence: 99%

Low PCSK9 levels are correlated with mortality in patients with end-stage liver disease

et al. 2017

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“…Cholesterol and cholesterol ester were determined based on our previously published LC-MS/MS method [17]. In brief, 10 μl of calibrator, quality control or sample (serum or plasma) was mixed with 490 μl internal standard solution (100 μg/L d7-cholesterol in methanol/isopropanol, 1/1, v/v) into polypropylene tubes.…”

Section: Methodsmentioning

confidence: 99%

Cholesterol esterification in plasma as a biomarker for liver function and prediction of mortality

et al. 2017

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BackgroundAdvanced stages of liver cirrhosis lead to a dramatically increased mortality. For valid identification of these patients suitable biomarkers are essential. The most important biomarkers for liver function are bilirubin and prothrombin time expressed as International Normalized Ratio (INR). However, the influence of several anticoagulants on the prothrombin time limits its diagnostic value.Aim of this study was the evaluation of cholesterol esterification (CE) fraction (esterified cholesterol vs. total cholesterol) as an alternative biomarker for liver synthesis and mortality prediction. Under physiological conditions the CE fraction in blood is closely regulated by lecithin-cholesterol acyltransferase (LCAT) which is produced in the liver.MethodsOne hundred forty-two patients with liver disease clinically considered for orthotopic liver transplant for different indications were enrolled in the study. One patient was excluded because of the intake of a direct oral factor Xa inhibitor which has a strong impact on prothrombin time.ResultsResults of CE fraction were in good agreement with INR (R2 = 0.73; p < 0.001). In patients who died or survived within three months mean CE fraction was 56% vs. 74% (p < 0.001) and mean INR was 2.0 vs. 1.3 (p < 0.001), respectively. The predictive value of CE fraction for three-month mortality risk was higher compared to INR (p = 0.04). Results for one-year mortality were comparable.ConclusionsThe cholesterol esterification fraction is a valid biomarker for liver synthesis and allows reliable prediction of mortality. In contrast to INR, it is independent of anticoagulation and other analytical limitations of coagulation tests.Electronic supplementary materialThe online version of this article (doi:10.1186/s12876-017-0614-9) contains supplementary material, which is available to authorized users.

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“…They were analyzed by LC–MS/MS (Lembcke et al 2005; Becker et al 2015). 25 µl of the supernatants were injected onto the analytical column (Chromolith SpeedROD RP-18e, 50 × 4.6 mm, monolithic column, Merck KGaA, Darmstadt, Germany).…”

Section: Methodsmentioning

confidence: 99%

Circulating sterols as predictors of early allograft dysfunction and clinical outcome in patients undergoing liver transplantation

et al. 2016

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IntroductionSensitive and specific assessment of the hepatic graft metabolism after liver transplantation (LTX) is essential for early detection of postoperative dysfunction implying the need for consecutive therapeutic interventions.ObjectivesHere, we assessed circulating liver metabolites of the cholesterol pathway, amino acids and acylcarnitines and evaluated their predictive value on early allograft dysfunction (EAD) and clinical outcome in the context of LTX.MethodsThe metabolites were quantified in the plasma of 40 liver graft recipients one day pre- and 10 days post-LTX by liquid chromatography/tandem mass spectrometry (LC–MS/MS). Plant sterols as well as cholesterol and its precursors were determined in the free and esterified form; lanosterol in the free form only. Metabolites and esterification ratios were compared to the model for early allograft function scoring (MEAF) which is calculated at day 3 post-LTX from routine parameters defining EAD.ResultsThe hepatic esterification ratio of all sterols, but not amino acids and acylcarnitine concentrations, showed substantial metabolic disturbances post-LTX and correlated to the MEAF. In ROC analysis, the low esterification ratio of β-sitosterol and stigmasterol from day 1 and of the other sterols from day 3 were predictive for a high MEAF, i.e. EAD. Additionally, the ratio of esterified β-sitosterol and free lanosterol were predictive for all days and the esterification ratio of the other sterols at day 3 or 4 post-LTX for 3-month mortality.ConclusionLow ratios of circulating esterified sterols are associated with a high risk of EAD and impaired clinical outcome in the early postoperative phase following LTX.Electronic supplementary materialThe online version of this article (doi:10.1007/s11306-016-1129-z) contains supplementary material, which is available to authorized users.

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LC–MS/MS-based quantification of cholesterol and related metabolites in dried blood for the screening of inborn errors of sterol metabolism

Cited by 24 publications

References 19 publications

Low PCSK9 levels are correlated with mortality in patients with end-stage liver disease

Low PCSK9 levels are correlated with mortality in patients with end-stage liver disease

Cholesterol esterification in plasma as a biomarker for liver function and prediction of mortality

Circulating sterols as predictors of early allograft dysfunction and clinical outcome in patients undergoing liver transplantation

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