Identification of prednisolone, methylprednisolone and their metabolites in human urine using HPLC (+) ESI-MS/MS and detection of possible adulteration in Indian herbal drug preparations

Abstract

Objective: To explore the possibility of identifying the maximum number of metabolites of prednisolone and methylprednisolonebyLC-MS/MSandtofurthertestthe applicationofthisdevelopedmethodonsixIndianherbal drug preparations. Method and Materials: The sample extraction procedure involves enzymatic hydrolysis and liquid-liquidextractionandfurtheranalysisusingLC-MS/ MS.Theexcretionprofilewasperformedwithfourhealthy male volunteers after administration of 40 mg and 8 mg ofprednisoloneandmethylprednisolone,respectively.Six herbaldrugpreparationsobtainedfromAllIndiaInstituteof MedicalSciences,Indiaweretestedtodetectthepossible adulteration with prednisolone or methylprednisolone. Results: The Analytical method was validated as per the requirement of WADA International Standard of Laboratories (version 6.0).The parent compound and ten urinary metabolites of prednisolone could be identified. The parent, M-1, M-2 and M-3 could be detected up to 72hourswhilerestofthemetabolitesweredetectableup to24hours.Inthecaseofmethylprednisolone,theparent compound and six urinary metabolites were identified. M-1 and M-2 of methylprednisolone were detectable up to 48 hours while the parent drug methylprednisolone andothermetabolitesweredetectableupto24hours.Out of the six herbal drugs tested, one showed the presence of prednisolone. Conclusion: The improved detection method developed for the identification and detection of prednisolone and methylprednisolone metabolites would provehighlybeneficialinextendingthetimeofdetectionof drugabuseinathletesandalsodetectingspikedayurvedic, homeopathicandunanipreparations.

Homologous ELISA for detection of prednisolone in human serum

“…The inter CV was within ±15% of the actual value. The coefficient of correlation was r 2 = 0.99 and LOD was 15 ng/ mL (Shobha et al, 2012). Leporati et al (2013) and Vincenti et al (2012) expanded and revalidated existing quantitative LC-MS/MS method for the detection of PSL in beef cattle (Leporati et al, 2013) and cow urine (Vincenti et al, 2012).…”

“…The limit of quantitation and CV% for PSL were 5.0 nmol/L and 7.6%, respectively (McWhinney et al, 2010). Shobha et al (2012) estimated prednisolone using HPLC (+) Electrospray ionization (ESI) -MS/MS method. The concentrations of calibration standard were 15-120 ng/mL.…”

“…It has been found that presence of Δ 1, 2 in corticosteroids enhances antiinflammatory activity and decreases salt-retaining activity (Delgado & Remers, 1998). PSL is often misused in sports due to its anti-inflammatory effect, which leads to a decrease in pain and increases an athlete's ability (Shobha et al, 2012). Similarly PSL improves pulmonary function in horse and is being used in equine sports to increase athlete horse performance (Fidani et al, 2012).…”