Simultaneous quantitation of seven endogenous C-21 adrenal steroids by liquid chromatography tandem mass spectrometry in human serum

Carvalho, Valdemir Melechco; Nakamura, Odete H.; Vieira, José Gilberto H.

doi:10.1016/j.jchromb.2008.07.035

Cited by 58 publications

(41 citation statements)

References 32 publications

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“…Intestinal P450 and DSS-Induced Colitis (Carvalho et al, 2008). Briefly, each tissue sample was prepared in 3 volumes of a methanol:water mixture (2:1, v/v).…”

Section: Methodsmentioning

confidence: 99%

Intestinal Epithelium–Specific Knockout of the Cytochrome P450 Reductase Gene Exacerbates Dextran Sulfate Sodium–Induced Colitis

Zhu

Xie

Ding

et al. 2015

J Pharmacol Exp Ther

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The potential involvement of intestinal microsomal cytochrome P450 (P450) enzymes in defending against colon inflammation and injury was studied in mice treated with dextran sulfate sodium (DSS) to induce colitis. Wild-type (WT) mice and mice with intestinal epithelium (IE)-specific deletion of the P450 reductase gene (IE-Cpr-null) were compared. IE-Cpr-null mice have little microsomal P450 activity in IE cells. DSS treatment (2.5% in drinking water for 6 days) caused more severe colon inflammation, as evidenced by the presence of higher levels of myeloperoxidase and proinflammatory cytokines [tumor necrosis factor-a, interleukin (IL)-6, and IL-1b], and greater weight loss, colonic tissue damage, and colon shortening, in IE-Cpr-null mice than in WT mice. The IE-Cpr-null mice were deficient in colonic corticosterone (CC) synthesis, as indicated by the inability of ex vivo cultured colonic tissues from DSS-treated IE-Cpr-null mice (in contrast to DSS-treated WT mice) to show increased CC production, compared with vehicle-treated mice, and by the ability of added deoxycorticosterone (DOC), a precursor of CC biosynthesis via mitochondrial CYP11B1, to restore ex vivo CC production by colonic tissues from DSStreated null mice. Intriguingly, null (but not WT) mice failed to show increased serum CC levels following DSS treatment. Nevertheless, cotreatment of DSS-exposed mice with DOC, which did not restore DSS-induced increase in serum CC, abolished the hypersensitivity of IE-Cpr-null mice to DSS-induced colon injury. Taken together, our results strongly support the notion that microsomal P450 enzymes in the intestine play an important role in protecting colon epithelium from DSS-induced inflammation and injury, possibly through increased local CC synthesis in response to DSS challenge.

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“…Intestinal P450 and DSS-Induced Colitis (Carvalho et al, 2008). Briefly, each tissue sample was prepared in 3 volumes of a methanol:water mixture (2:1, v/v).…”

Section: Methodsmentioning

confidence: 99%

Intestinal Epithelium–Specific Knockout of the Cytochrome P450 Reductase Gene Exacerbates Dextran Sulfate Sodium–Induced Colitis

Zhu

Xie

Ding

et al. 2015

J Pharmacol Exp Ther

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“…In contrast, in routine clinical LC-MS/MS applications, this approach is only rarely used at present (41,43,44 ). Besides the fact that, for many analytes, collision-induced dissociation generates only a single product ion of sufficient intensity to be detected, this technique is not widely used, mainly because of the practical reasons described below.…”

Section: Pitfalls Of Lc-ms/msmentioning

confidence: 99%

“…It is possible that the disintegration patterns of a pair of isomers may be different, which would therefore allow for analytical discrimination between the 2 compounds, but in most cases, the chromatographic baseline separation of an analyte and its isomer before their MS/MS detection is required for unequivocal quantitative measurements to be performed (44 ) (Fig. 4).…”

Section: Pitfalls Of Lc-ms/msmentioning

confidence: 99%

Pitfalls Associated with the Use of Liquid Chromatography–Tandem Mass Spectrometry in the Clinical Laboratory

2010

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BACKGROUND Novel mass spectrometric techniques such as atmospheric pressure ionization and tandem mass spectrometry have substantially extended the spectrum of clinical chemistry methods during the past decade. In particular, liquid chromatography tandem–mass spectrometry (LC-MS/MS) has become a standard tool in research laboratories as well as in many clinical laboratories. Although LC-MS/MS has features that suggest it has a very high analytical accuracy, potential sources of inaccuracy have recently been identified. CONTENT The sources of inaccuracy in LC-MS/MS methods used in the routine quantification of small molecules are described and discussed. Inaccuracy of LC-MS/MS methods can be related to the process of ionization through the insource transformation of conjugate metabolites or target analytes and may also be attributable to ionization matrix effects that have a differential impact on target analytes and internal-standard compounds. Inaccuracy can also be associated with the process of ion selection, which mainly occurs when compounds from the sample matrix share mass transitions with a target analyte. In individual assays, most potential sources of inaccuracy can be controlled by sufficient LC separation–based sample workup before MS analysis. SUMMARY LC-MS/MS methods should undergo rigorous and systematic validation before introduction into patient care.

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“…The reliable and simultaneous analysis of steroid hormones panel is a powerful tool for investigation of steroid hormone status, which is relevant for the diagnosis and treatment of a variety of clinical disorders like adrenal insufficiency and congenital adrenal hyperplasia (CAH) [1][2][3][4][5]. Due to the low levels of serum steroids metabolites and chemical similarity, analysis of this kind of compounds has been a challenge [1].…”

Section: Introductionmentioning

confidence: 99%

“…Due to the low levels of serum steroids metabolites and chemical similarity, analysis of this kind of compounds has been a challenge [1]. Historically androgens were tested in urine using colorimetric assays [2].…”

Section: Introductionmentioning

confidence: 99%

Analytical Performance of LC-MS/MS Method for Simultaneous Determination of Five Steroids in Serum

Alves¹

2015

Mass Spectrom Purif Tech

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The accurate measurement of steroid hormones is the fulcrum of modern endocrinology. Our aim was to develop a mass spectrometry coupled with a liquid chromatographic method for 17-hydroxyprogesterone, androstenedione, 11-desoxycortisol, 21-desoxycortisol and cortisol serum quantification simultaneously. The samples were submitted to isotopic internal standards addition, protein precipitation and two dimensional liquid chromatography consisting of trapping column and reverse-phase C18 analytical column following atmospheric pressure chemical ionization and mass spectrometry detection. For all compounds tested functional sensitivity was less than 0.5 ng/mL, precision was less than 15%, recovery ranged from 93% to 120%, linearity ranged from 89% to 111%, and accuracy was considered adequate. In conclusion, we developed a suitable, fast and reliable method for simultaneous routine measurement of 17-hydroxyprogesterone, androstenedione, 11-desoxycortisol, 21-desoxycortisol and cortisol in serum.

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Simultaneous quantitation of seven endogenous C-21 adrenal steroids by liquid chromatography tandem mass spectrometry in human serum

Cited by 58 publications

References 32 publications

Intestinal Epithelium–Specific Knockout of the Cytochrome P450 Reductase Gene Exacerbates Dextran Sulfate Sodium–Induced Colitis

Intestinal Epithelium–Specific Knockout of the Cytochrome P450 Reductase Gene Exacerbates Dextran Sulfate Sodium–Induced Colitis

Pitfalls Associated with the Use of Liquid Chromatography–Tandem Mass Spectrometry in the Clinical Laboratory

Analytical Performance of LC-MS/MS Method for Simultaneous Determination of Five Steroids in Serum

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