Hydrolysis of Pregnanediol Glucuronide by Sodium Periodate

Wade, A. P.

doi:10.1042/bj1030019c

Cited by 4 publications

(3 citation statements)

References 7 publications

(6 reference statements)

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Section: Methodsmentioning

confidence: 99%

“…For the hydrolysis of the glucuronides which remain in urine after β-glucuronidase treatment, a chemical hydrolysis was performed on the basis of a previous described procedure. 12 The method was applied to the fractions collected after LC purification. Briefly, the residue of the LC fraction was dissolved in 2 mL of an aqueous solution of sodium metaperiodate (10% w/v), and the pH was adjusted to a value of 3 with an 1 M hydrochloric acid solution.…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

“…Chemical hydrolysis approaches are also available to hydrolyze glucuronoconjugates. , However, they also present some limitations such as degradation of some analytes and an increase in matrix interferences.…”

mentioning

confidence: 99%

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Use of LC-MS/MS for the Open Detection of Steroid Metabolites Conjugated with Glucuronic Acid

et al. 2013

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In humans, conjugation with glucuronic acid is the most important phase II metabolic reaction of steroidal compounds. Glucuronoconjugated metabolites have been conventionally studied by using β-glucuronidase enzymes to release the phase I metabolites. It is well-known that hydrolysis with β-glucuronidase presents some limitations that may result in the underestimation of some conjugates. The aim of the present work was to develop and to evaluate liquid chromatography-tandem mass spectrometry (LC-MS/MS) scan methods for the open detection of steroid glucuronides in urine samples. The mass spectrometric behavior of thirteen representative steroid glucuronides, used as model compounds, was studied. Characteristic ionization and collision induced dissociation behaviors were observed depending on the steroid glucuronide structure. Neutral loss (NL of 176, 194, 211, and 229 Da) and precursor ion (PI of m/z 141, 159, and 177, in positive mode and m/z 75, 85, and 113, in negative mode) scan methods were evaluated. The NL scan method was chosen for the open detection of glucuronoconjugated steroids due to its sensitivity and the structural information provided by this method. The application of the NL scan method to urine samples collected after testosterone (T) undecanoate administration revealed the presence of two T metabolites which remain conjugated as glucuronides after an enzymatic hydrolysis of the urine. 3α,6β-Dihydroxy-5α-androstan-17-one (6β-hydroxyandrosterone) glucuronide and 3α,6β-dihydroxy-5β-androstan-17-one (6β-hydroxyetiocholanolone) glucuronide were established as the structures for these metabolites, by comparing the structure of the steroids released after chemical hydrolysis with reference materials. An increase of 50-300-fold of these metabolites after oral administration of T undecanoate was observed, proving that their determination can be useful in the doping control field. Moreover, these results exemplify that significant information might be missed, unless direct methods for the determination of steroid glucuronides are employed.

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Section: Methodsmentioning

confidence: 99%

Section: ■ Experimental Sectionmentioning

confidence: 99%