A Novel Liquid Chromatography Tandem Mass Spectrometry Method for the Estimation of Bilirubin Glucuronides and its Application to In Vitro Enzyme Assays

Putluru, Siva P; Matta, Murali K.; Ahire, Deepak; Subramanian, Murali; Sinz, Michael; Mandlekar, Sandhya

doi:10.2174/1872312810666161124143522

Cited by 12 publications

(6 citation statements)

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“…We found more intense peaks in positive mode when compare to the negative mode using ESI source. MRM technique was used to monitor the ions for better selectivity and specificity [12][13][14][15] . Mass spectrometry source and compound dependent parameters were suitably tuned to get better intense signals and reproducible response.…”

Section: Resultsmentioning

confidence: 99%

Bioanalysis of Febuxostat in Human Plasma by Liquid Chromatography/Tandem Mass Spectrometry

Kovvasu¹,

Yeung²,

Kunamaneni³

et al. 2018

IJPTR

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Abstract:The objective of this paper was to describe the development and validation of a novel liquid chromatography / tandem mass spectrometry (LC-MS/MS) method for the determination of Febuxostat in human plasma using febuxostat-d9 as internal standard (IS). An API-4000 triple quadrupole mass spectrometer operated in MRM mode was used for the selective quantification of Febuxostat in human plasma. Sample extraction utilizes protein precipitation (PP), followed by analyte and the IS were chromatographed on a C18 column using an isocratic mobile phase composed of 5mM ammonium formate and acetonitrile (20:80, v/v) pumped at a flow rate of 1.0 mL/min. Precision and accuracy of the method was determined using five analytical batches in the concentration range of 15.0-8000 ng/mL. All the validation experiments were carried out as per the US FDA guidelines and results met the acceptance criteria. The developed method was rapid with a total chromatographic run time of 2.0 min.

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Section: Resultsmentioning

confidence: 99%

Bioanalysis of Febuxostat in Human Plasma by Liquid Chromatography/Tandem Mass Spectrometry

Kovvasu¹,

Yeung²,

Kunamaneni³

et al. 2018

IJPTR

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“…Blank serum was substituted by HCl (1.5%). The values were measured in mg/dl of serum (Putluru et al, 2016).…”

Section: Estimation Of Bilirubinmentioning

confidence: 99%

Effect of Katakakhadiradi kashayam on liver metabolism in streptozotocin and nicotinamide induced diabetic rats

Subbiah¹,

Kavimani²,

Ramadoss³

et al. 2021

ijrps

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Plants have the ability to impart therapeutic action on chronic diseases such as diabetes and its comorbidities. The current study was done to find the effectiveness of Katakakhadiradi kashayam on liver metabolism in streptozotocin and nicotinamide administered diabetic rats. Diabetic rats were treated with katakakhadiradi kashayam for 28 days and compared with glibenclamide. The antidiabetic action of Katakakhadiradi kashayam was measured by the serum levels of urea, bilirubin, SGOT and SGPT activity. In the current study, the level of urea, SGPT and SGOT activity was significantly higher and reduced total bilirubin observed in diabetic rats. The protection given by Katakakhadiradi kashayam against diabetic complication was confirmed by a histology study. Most of the plants present in Kadakakhadiradi kashayam are effective in regulating the enzymes in diabetes mellitus. The obtained data imply the antidiabetic effects of katakakhadiradi kashayam, which is practically a safe herbal formulation and can be used as a good alternative for managing DM.

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“…Based on LC‐MS/MS analysis, bilirubin has been previously determined as a selective substrate of UGT1A1, [7] however, the instability and photodecomposition of bilirubin restricts its application in detection of UGT1A1 activity within biological specimens. With the development of fluorescence‐based technologies, the many advantages including high sensitivity, [8] real‐time detection, excellent spatiotemporal resolution and non‐invasiveness as well as high‐throughput screening capacity, have provided a route to break the bottleneck associated with traditional assay methods [9] .…”

Section: Introductionmentioning

confidence: 99%

A Molecular‐Splicing Strategy for Constructing a Near‐Infrared Fluorescent Probe for UDP‐Glucuronosyltransferase 1A1

Tian

Liu

et al. 2021

Angewandte Chemie

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UDP-glucuronosyltransferase 1A1 (UGT1A1) is av ital metabolic enzyme responsible for the clearance of endogenous substances and drugs.H itherto,t he development of fluorescent probes for UGTs was severely restricted due to the poor isoform selectivity and on-off or blue-shifted fluorescence response.Herein, we established anovel "molecular-splicing" strategy to construct ah ighly selective nearinfrared (NIR) fluorescent probe, HHC,f or UGT1A1, which exhibited aN IR signal at 720 nm after UGT1A1 metabolism. HHC was then successfully used for the real-time imaging of endogenous UGT1A1 in living cells and animals and to monitor the bile excretion function. In summary,a ni soformspecific NIR fluorescent probe has been developed for monitoring UGT1A1 activity in living systems,h igh-throughput screening of novel UGT1A1 inhibitors and visual evaluation of bile excretion function.

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A Novel Liquid Chromatography Tandem Mass Spectrometry Method for the Estimation of Bilirubin Glucuronides and its Application to In Vitro Enzyme Assays

Abstract: We have developed a sensitive LC-MS/MS based method for the quantitation of bilirubin and its glucuronides from in vitro incubations. This method was successfully utilized to determine bilirubin glucuronidation kinetics in HLM and human rUGT1A1.

Cited by 12 publications

References 0 publications

Bioanalysis of Febuxostat in Human Plasma by Liquid Chromatography/Tandem Mass Spectrometry

Bioanalysis of Febuxostat in Human Plasma by Liquid Chromatography/Tandem Mass Spectrometry

Effect of Katakakhadiradi kashayam on liver metabolism in streptozotocin and nicotinamide induced diabetic rats

A Molecular‐Splicing Strategy for Constructing a Near‐Infrared Fluorescent Probe for UDP‐Glucuronosyltransferase 1A1

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