Simultaneous quantitation of fatty acids, sterols and bile acids in human stool by capillary gas–liquid chromatography

Batta, Ashok K.; Salen, Gerald; Batta, Priti; Tint, G. S.; Alberts, David S.; Earnest, David L.

doi:10.1016/s1570-0232(02)00289-1

Cited by 71 publications

(50 citation statements)

References 14 publications

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“…The dried extract was saponified by adding freshly prepared 1 N KOH ethanolic solution (3 ml). Total cholesterol was derivatized as described previously (26). Recoveries for cholesterol and cholesteryl esters were assessed using cholesterol and cholesteryl oleate standards and found to range from 84% to 92%.…”

Section: Cholesterol Extraction and Derivatizationmentioning

confidence: 99%

Assessing the effects of LXR agonists on cellular cholesterol handling: a stable isotope tracer study

Webb

Jaye

Ghosh

et al. 2006

Journal of Lipid Research

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The liver X receptors (LXRs) a and b are responsible for the transcriptional regulation of a number of genes involved in cholesterol efflux from cells and therefore may be molecular targets for the treatment of cardiovascular disease. However, the effects of LXR ligands on cholesterol turnover in cells has not been examined comprehensively. In this study, cellular cholesterol handling (e.g., synthesis, catabolism, influx, and efflux) was examined using a stable isotope labeling study and a two-compartment modeling scheme. In HepG2 cells, the incorporation of 13 C into cholesterol from [1-13 C]acetate was analyzed by mass isotopomer distribution analysis in conjunction with nonsteady state, multicompartment kinetic analysis to calculate the cholesterol fluxes. Incubation with synthetic, nonsteroidal LXR agonists (GW3965, T0901317, and SB742881) increased cholesterol synthesis (z10-fold), decreased cellular cholesterol influx (71-82%), and increased cellular cholesterol efflux (1.7-to 1.9-fold) by 96 h. As a consequence of these altered cholesterol fluxes, cellular cholesterol decreased (36-39%) by 96 h. The increased cellular cholesterol turnover was associated with increased expression of the LXR-activated genes ABCA1, ABCG1, FAS, and sterol-regulatory element binding protein 1c.In summary, the mathematical model presented allows time-dependent calculations of cellular cholesterol fluxes. These data demonstrate that all of the cellular cholesterol fluxes were altered by LXR activation and that the increase in cholesterol synthesis did not compensate for the increased cellular cholesterol efflux, resulting in a net cellular cholesterol loss.-Aravindhan, K., C. L. Webb, M. Jaye, A. Ghosh, R. N. Willette, N. J. Di Nardo, and B .M. Jucker. The liver X receptor (LXR) is an oxysterol-sensing nuclear receptor responsible for transcriptional regulation of a number of genes involved in reverse cholesterol transport and therefore may be a molecular target for the treatment of cardiovascular disease (1-3). LXR is a member of the nuclear receptor family that regulates cellular cholesterol efflux (4) and whole body cholesterol excretion (5) and is endogenously activated by various oxysterols (6), including 24(S),25-epoxycholesterol and 22(R)-hydroxycholesterol, an intermediate in steroid hormone production (7, 8). Additionally, synthetic LXR agonists that potently upregulate macrophage ABCA1 expression have been shown to be antiatherogenic in genetic mouse models of atherosclerosis (9, 10).Cellular cholesterol metabolism and handling has been examined in terms of its synthesis by measuring the fraction of newly synthesized cholesterol (11,12), in terms of catabolism by measuring levels of bile acids through biochemical assays or HPLC (13,14), in terms of influx by measuring the radioactivity of absorbed [ 3 H]cholesteryl oleate (15, 16), and in terms of efflux by measuring specific radioactivity in labeled cells (17)(18)(19)(20). However, prior methods address only the unidirectional flow of cholesterol and are not ...

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Section: Cholesterol Extraction and Derivatizationmentioning

confidence: 99%

Assessing the effects of LXR agonists on cellular cholesterol handling: a stable isotope tracer study

Webb

Jaye

Ghosh

et al. 2006

Journal of Lipid Research

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“…The fecal neutral sterols were analysed in freeze-dried samples by gas chromatography on a 25-m fused silica capillary column (CP-SIL 5CB, 0.25-mm i.d., and 0.25-µm film thickness; Varian, U.S.A.). The samples were then subjected to nbutylation and trimethylsilylation (Sylon HTP was purchased from Supelco), according to Batta et al (2002), except that the reaction mixture was centrifuged prior to evaporation at 1400 g for 10 min. The residue was dissolved in hexane and centrifuged, and then a 1-µL sample was injected.…”

Section: Chemical Analysesmentioning

confidence: 99%

Hydrophobically Modified Celluloses as Novel Cholesterol-lowering Polymers

Tůma¹,

Volek²,

Synytsya³

et al. 2014

BioResources

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Hydrophobically modified celluloses were prepared by the esterification of monocarboxycellulose (MCC) and carboxymethylcellulose (CMC) with methanol, followed by the amino-de-alkoxylation of the methyl esters with n-octadecylamine. These cellulose derivatives were fed at 30 and 60 g/kg for 4 weeks to female rats. The diets were supplemented with palm fat (60 or 50 g/kg) and cholesterol (0 or 10 g/kg). There was no significant effect of amidated celluloses on the feed intake or body weight of rats. Both MCC and CMC derivatives significantly decreased the concentrations of cholesterol present in the serum and liver. The supplementation of diets with hydrophobically modified celluloses tended to increase fecal concentrations of cholesterol and coprostanol and significantly increased fecal concentrations of total neutral sterols. In rats fed diets containing cholesterol, the total serum cholesterol correlated negatively with the fecal concentrations of neutral sterols. It can be concluded that MCC-C18 and CMC-C18 are effective cholesterollowering agents.

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“…The silyl derivates were quantified on a gas chromatograph equipped with an SAC-5 capillary column (Supelco, Bellefonte, U.S.A.), operated isothermally at 285°C. The neutral and acidic faecal sterols were determined in freeze-dried samples by gas chromatography on a capillary column CP-SIL 5CB (Varian, U.S.A.) after butylation and silylation according to Batta et al [16]. Norcholic acid was used as the internal standard.…”

Section: Analysesmentioning

confidence: 99%

Gender-based differences in the effect of dietary cholesterol in rats

et al. 2012

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Male and female rats were fed diets supplemented with cholesterol and palm fat at 10 and 50 g/kg, respectively; serum, hepatic tissue and faeces were analysed. Cholesterol supplementation significantly increased serum and hepatic cholesterol both in male and female rats. Male and female rats fed the cholesterol-containing diet differed significantly in serum cholesterol concentration (2.48 µmol/mL vs 2.92 µmol/mL), concentration of serum triacylglycerols, but not in hepatic cholesterol concentration. The serum and hepatic cholesterol concentrations correlated non-significantly in male rats (r=0.491; P=0.063) and significantly in female rats (r=0.818; P<0.001). Cholesterol supplementation non-significantly decreased relative expression of the hepatic LDL receptor gene and significantly increased relative expression of the hepatic cholesterol 7α-hydroxylase gene in rats of both genders. The faeces of control rats contained similar amounts of cholesterol and bile acids. Cholesterol supplementation increased cholesterol concentration 10 times in the faeces of male rats and 12 times in faeces of female rats. The corresponding increases of bile acid concentration were much lower (83% in male rats and 108% in female rats). It can be concluded that the effects of cholesterol supplementation were more pronounced in female than in male rats.

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Simultaneous quantitation of fatty acids, sterols and bile acids in human stool by capillary gas–liquid chromatography

Cited by 71 publications

References 14 publications

Assessing the effects of LXR agonists on cellular cholesterol handling: a stable isotope tracer study

Assessing the effects of LXR agonists on cellular cholesterol handling: a stable isotope tracer study

Hydrophobically Modified Celluloses as Novel Cholesterol-lowering Polymers

Gender-based differences in the effect of dietary cholesterol in rats

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