Determination of raloxifene and its glucuronides in human urine by liquid chromatography–tandem mass spectrometry assay

Trdan, Tina; Roškar, Robert; Trontelj, Jurij; Ravnikar, M.; Mrhar, Aleš

doi:10.1016/j.jchromb.2011.06.031

Cited by 27 publications

(30 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…ATCC 55043 [20] followed by semipreparative chromatographic purification and lyophilisation. The details of the synthesis and purification are described elsewhere [21]. Purity was checked by HPLC and LC-MS/MS.…”

Section: Methodsmentioning

confidence: 99%

Influence of hepatic and intestinal efflux transporters and their genetic variants on the pharmacokinetics and pharmacodynamics of raloxifene in osteoporosis treatment

Lušin

Mrhar

Stieger

et al. 2012

Translational Research

Self Cite

View full text Add to dashboard Cite

Raloxifene exhibits a large and unexplained interindividual variability in its pharmacokinetics and pharmacodynamics. The aim of our study was to identify transporters involved in the efflux of raloxifene and its glucuronide metabolites by various in vitro models and by an in vivo study to explore the possible involvement of P-glycoprotein (Pgp), multidrug resistance-associated protein (MRP)1, MRP2, MRP3, and breast cancer resistance protein in the observed high interindividual variability. Experiments with the parallel artificial membrane permeability assay showed the highest passive permeability for raloxifene, followed by raloxifene-6--glucuronide (M1), raloxifene-4'--glucuronide (M2), and raloxifene-6,4'-diglucuronide (M3). Caco-2 cell monolayer experiments indicated an interaction of raloxifene with Pgp. The ATPase assay confirmed the raloxifene interaction with Pgp and indicated interactions of all raloxifene species with MRP1, MRP2, MRP3, and breast cancer resistance protein, except for M1, which did not show any interactions with MRP2. Furthermore, the vesicular experiments confirmed the interaction of M2 and M3 with MRP2. Although the in vivo study on osteoporotic postmenopausal women on raloxifene could not confirm a significant influence of ABCB1 and ABCC2 genetic polymorphisms on its pharmacokinetics, a clear trend toward higher total raloxifene concentrations was observed in carriers of at least 1 ABCB1 c.3435T allele. Moreover, the same polymorphism effect was also observed as a significant increase in total hip bone mineral density after 1 year of treatment. The results of our study support the involvement of efflux transporters in disposition of raloxifene and its metabolites and may partially explain the observed raloxifene variability by the influence of the ABCB1 c.3435C>T polymorphism. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 2 AbstractRaloxifene exhibits quite a large and unexplained interindividual variability in its pharmacokinetics and pharmacodynamics. The aim of our study was to identify transporters involved in the efflux of raloxifene and its glucuronide metabolites by various in vitro models and by an in vivo study to explore the possible involvement of Pgp, MRP1, MRP2, MRP3, and BCRP in the observed high interindividual variability. Experiments with PAMPA (Parallel Artificial Membrane Permeability Assay) showed the highest passive permeability for raloxifene, followed by raloxifene-6-β-glucuronide (M1), raloxifene-4'-β-glucuronide (M2) and raloxifene-6,4'-diglucuronide (M3). Caco-2 cell monolayer experiments indicated an inte...

show abstract

Section: Methodsmentioning

confidence: 99%

Influence of hepatic and intestinal efflux transporters and their genetic variants on the pharmacokinetics and pharmacodynamics of raloxifene in osteoporosis treatment

Lušin

Mrhar

Stieger

et al. 2012

Translational Research

Self Cite

View full text Add to dashboard Cite

show abstract

“…As example for this approach, the elucidation of three raloxifene glucuronides in urine as well as their identity confirmation after bioproduction by using QQQ is provided [37]. Chromatograms of each bioproduced glucuronide standard obtained in ESI positive full scan mode gave only one chromatographic peak where MS spectra of each peak showed strong molecular ions at m/z 650, 650 and 826 for two raloxifene monoglucuronides and diglucuronide, respectively.…”

Section: Examples Of Metabolite Identificationmentioning

confidence: 99%

“…If the conversion is complete, this approach is valid for determination of the metabolite stock solution concentrations when small amounts of glucuronide standards are obtained or available [37].…”

Section: Examples Of Metabolite Identificationmentioning

confidence: 99%

Analytical Methods for Quantification of Drug Metabolites in Biological Samples

Roškar¹,

Lušin²

2012

Chromatography - The Most Versatile Method of Chemical Analysis

Self Cite

View full text Add to dashboard Cite

“…Determination of RLX in its formulations with a method based on measurement of the intensity of resonance Rayleigh scattering of the ion association complex of RLX with Evans blue was reported 26 (LOQ = 60.0 ng mL -1 ) but it requires an expensive instrumental set up. On the other side, liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been successfully applied for determination of RXL in human urine (LOQ = 0.515 to 20.0 ng mL -1 ) 27,28 and in human plasma (LOQ = 20.00 ng L -1 ). 29 However, LC-MS/MS methods involve expensive solid phase extraction steps prior to the analysis which are time-consuming and not economically feasible for routine analysis (as such equipment and techniques are not available in most of the laboratories).…”

Section: Introductionmentioning

confidence: 99%

“…29 However, LC-MS/MS methods involve expensive solid phase extraction steps prior to the analysis which are time-consuming and not economically feasible for routine analysis (as such equipment and techniques are not available in most of the laboratories). Moreover, most of the methods reported [9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28] are considered not efficient enough for the assay of RLX in human plasma and at different therapeutic dose levels for pharmacokinetic studies as well as therapeutic drug monitoring. Therefore, a simple and sensitive procedure is desired for determination of RLX in human plasma.…”

Section: Introductionmentioning

confidence: 99%

Adsorptive stripping voltammetry determination of the hormone therapy Raloxifene HCl in formulation and human serum

Ghoneim¹,

Hassanein²,

Salahuddin³

et al. 2012

J. Braz. Chem. Soc.

View full text Add to dashboard Cite

Raloxifeno HCl (RLX) adsorveu fortemente na superfície do eletrodo de mercúrio numa monocamada com cobertura superficial de 5,724 × 10 −10 mol cm -2 . Dois métodos voltamétricos de redissolução catódico adsortivo com varredura linear e onda quadrada (LS-AdCSV e SW-AdCSV, respectivamente) precisos, rápidos e livres de extração são descritos para determinação em nível de traço de RLX em solução, em formulação comercial e soro humano. Limites de quantificação (LOQ) de 2,0 × 10 -9 e 5,0 × 10 -11 mol L −1 de RLX em solução e 4,0 × 10 -9 e 1,0×10 -10 mol L −1 em soro humano enriquecido foram obtidos pelos métodos descritos LS-AdCSV e SW-AdCSV, respectivamente. Interferências de alguns excipientes comuns, íons metálicos e drogas coadministradas foram insignificantes. LOQ adquidos pelos métodos são baixos assim como oferecem boas possibilidades de determinação de drogas em preparações farmacológicas de baixa dosagem. Contudo, o método SW-AdCSV descrito é sensível suficiente em ensaios de drogas também em soro humano.Raloxifene HCl (RLX) was found to strongly adsorb onto surface of the mercury electrode in a monolayer surface coverage of 5.724 × 10 −10 mol cm -2 . Two precise, rapid and extractionfree linear sweep and square wave adsorptive cathodic stripping voltammetry (LS-AdCSV and SW-AdCSV, respectively) methods are described for trace quantitation of RLX in bulk form, commercial formulation and human serum. Limits of quantification (LOQ) of 2.0 × 10 -9 and 5.0 × 10 -11 mol L −1 RLX in bulk form and 4.0 × 10 -9 and 1.0 × 10 -10 mol L −1 RLX in spiked human serum were achieved by the described LS-AdCSV and SW-AdCSV methods, respectively. Insignificant interferences from some common excipients, metal ions and co-administrated drugs were obtained. LOQ achieved by the methods are low as well as they offer good possibilities for determination of drug in low-dosage pharmaceutical preparations. However, the SW-AdCSV method described is sensitive enough to assay the drug also in human serum.

show abstract

Determination of raloxifene and its glucuronides in human urine by liquid chromatography–tandem mass spectrometry assay

Cited by 27 publications

References 18 publications

Influence of hepatic and intestinal efflux transporters and their genetic variants on the pharmacokinetics and pharmacodynamics of raloxifene in osteoporosis treatment

Influence of hepatic and intestinal efflux transporters and their genetic variants on the pharmacokinetics and pharmacodynamics of raloxifene in osteoporosis treatment

Analytical Methods for Quantification of Drug Metabolites in Biological Samples

Adsorptive stripping voltammetry determination of the hormone therapy Raloxifene HCl in formulation and human serum

Contact Info

Product

Resources

About