High-performance liquid chromatographic method for the direct determination of 4-methylumbelliferone and its glucuronide and sulfate conjugates

Zimmerman, Cheryl L.; Ratna, Sanjeev; Leboeuf, Elizabeth; Pang, K. Sandy

doi:10.1016/0378-4347(91)80279-l

Cited by 20 publications

(15 citation statements)

References 10 publications

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“…In order to determine the purity of peaks during preliminary separations, chromatographic runs were made by measuring the absorbance from 200 nm to 400 nm using the diode array scanning. From the absorbance profile, the D Vigetti et al maximum peak at 7 min was 313 nm according to a previous study (Zimmerman et al 1991) and was identified as 4-MUG.…”

Section: Analysis Of 4-mu Glucuronide (4-mug) By Hplcmentioning

confidence: 99%

“…Chromatographic separation was done with a Nucleosil R C18 analytical column (4.6 mm i.d., 25 cm length; Macherey-Nagel, Düren, Germany) at room temperature, as described by Zimmerman et al (1991), with minor modifications. The column was equilibrated with a KH 2 PO 4 50 mM methanol solution (80/20; v/v), filtered through a 0.22 μm membrane filter.…”

Section: Analysis Of 4-mu Glucuronide (4-mug) By Hplcmentioning

confidence: 99%

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The effects of 4-methylumbelliferone on hyaluronan synthesis, MMP2 activity, proliferation, and motility of human aortic smooth muscle cells

Vigetti¹,

Rizzi²,

Viola³

et al. 2009

Glycobiology

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Extracellular matrix remodeling after proatherosclerotic injury involves an increase in hyaluronan (HA) that is coupled with vascular smooth muscle cell (SMC) migration, proliferation, and with neointima formation. As such events are dependent on HA, in this study we assessed the effects on SMC behavior of 4-methylumbelliferone (4-MU). As previously described in other cell types, 4-MU reduced HA in cultures of primary human aortic SMCs (AoSMCs) as well as the cellular content of the HA precursor UDP-glucuronic acid. We found that SMCs increased UDP-glucuronyl transferase 1 enzymes, which can reduce the cellular content of UDP-glucuronic acid confirming that the availability of the UDP-sugar substrates can regulate HA synthesis. Interestingly, we reported that 4-MU reduced the transcripts coding for the three HA synthases as well as UDP glucose pyrophosphorylase and dehydrogenase. As HA synthase transcript reduction is common to other cell types, the 4-MU effect on gene expression may be considered a mechanism for HA synthesis inhibition. Moreover, we showed that 4-MU strongly inhibits AoSMCs migration, which was restored by the addition of exogenous HA indicating that the rescuing depends on the interaction of HA with its receptor CD44. Besides the decrease in HA synthesis and cell migration, 4-MU reduced AoSMCs proliferation, indicating that 4-MU may exert a vasoprotective effect.

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Section: Analysis Of 4-mu Glucuronide (4-mug) By Hplcmentioning

confidence: 99%

Section: Analysis Of 4-mu Glucuronide (4-mug) By Hplcmentioning

confidence: 99%

The effects of 4-methylumbelliferone on hyaluronan synthesis, MMP2 activity, proliferation, and motility of human aortic smooth muscle cells

Vigetti¹,

Rizzi²,

Viola³

et al. 2009

Glycobiology

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“…From this standpoint, we performed in situ intestinal perfusional experiments in Eisai hyperbilirubinemic rats (EHBRs), Bcrp1 (Ϫ/Ϫ) mice, and corresponding normal rats and mice, respectively. As test substances, we chose 4-methyl-umbelliferone (4MU) and E3040 which are metabolized in enterocytes by UDP-glucuronosyl transferases and sulfotransferases to yield conjugated metabolites (Mulder et al, 1985;Zimmerman et al, 1991;Takenaka et al, 1995a,b;Chen and Pang, 1997).…”

mentioning

confidence: 99%

Role of Breast Cancer Resistance Protein (Bcrp1/Abcg2) in the Extrusion of Glucuronide and Sulfate Conjugates from Enterocytes to Intestinal Lumen

Adachi¹,

Suzuki²,

Schinkel³

et al. 2004

Mol Pharmacol

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The purpose of this study is to examine the significance of efflux transporters in the small intestine to extrude glucuronide (G) and sulfate (S) conjugates into the intestinal lumen. From this standpoint, we performed in situ intestinal perfusion experiments by using Eisai hyperbilirubinemic rats (EHBRs) in which the multidrug resistance protein 2 (Mrp2/Abcc2) is hereditarily defective and breast cancer resistance protein (Bcrp1/Abcg2) knockout mice. The intestinal lumen of EHBRs and Bcrp1 (Ϫ/Ϫ) mice was perfused with medium containing 4-methylumbelliferone (4MU) and E3040 [6-hydroxy-5,7-dimethyl-2-methylamino-4-(3-pyridilmethyl) benzothiazole] to determine the efflux of metabolites into the outflow.

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“…21 The assay of 4MUS in S9 was similar to that for the perfusate samples. After the addition of 10 µL of the internal standard, p-nitrophenol (100 µg/mL) and 350 µL of methanol, the supernatant was transferred and dried under nitrogen.…”

Section: Mus and Metabolite Assaymentioning

confidence: 99%

“…21 The regions associated with the authentic standards (4MUS, 4MU, and 4MUG [and their radiolabeled counterparts]) were visualized under ultraviolet light and scraped into a scintillation vial. After the addition of 0.5 mL water and 10 mL scintillation fluor (Ready Protein, Beckman), the radioactivity of each sample was assayed by liquid scintillation counting (Beckman Canada, Model 5801 Liquid scintillation counter, Mississauga, Ontario, Canada).…”

Section: Mus and Metabolite Assaymentioning

confidence: 99%

Carrier-mediated entry of 4-methylumbelliferyl sulfate: Characterization by the multiple-indicator dilution technique in perfused rat liver

Chiba¹,

Schwab²,

Goresky³

et al. 1998

Hepatology

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The hepatocellular entry of 4-methylumbelliferyl sulfate (4MUS) a highly ionized and highly bound anion capable of futile cycling, was examined in the single-pass albumin-free perfused rat liver preparation. Desulfation of 4MUS to 4-methylumbelliferone (4MU) was verified in vitro to be a low-affinity, high-capacity process (Km = 731 micromol/L; Vmax = 414 nmol min(-1) g(-1) liver). With 4MUS given to the perfused rat liver, sulfation of 4MU, the formed metabolite, was attenuated in the presence of 2,6-dichloro-4-nitrophenol (DCNP), a sulfation inhibitor, and when sulfate ion was substituted by chloride ion. 4MU sulfation, being a high-affinity system, was reduced most effectively at the lowest 4MUS concentration (15 micromol/L) used, evidenced by the increased (24%) net hepatic extraction ratio of 4MUS and reduced utilization (72%) of infused tracer 35SO4(2-) by 4MU for 4MU35S formation. Single-pass multiple indicator dilution (MID) studies were thus conducted under identical conditions (DCNP and absence of inorganic sulfate), with injection of [3H]4MUS and a set of noneliminated vascular and cellular reference indicators into the portal vein (prograde) or hepatic vein (retrograde), against varying background bulk concentrations of 4MUS (5 to 900 micromol/L). The steady-state removal rate of 4MUS and formation rates of 4MU and its glucuronide conjugate (4MUG) were not altered with perfusion flow direction, suggesting the presence of even or parallel distributions of 4MUS desulfation and 4MU glucuronidation activities. When the outflow dilution profile of [3H]4MUS was evaluated with the barrier-limited model of Goresky, a slight red cell carriage effect was found for 4MUS. The permeability surface area product for cellular entry for prograde showed a dramatic concentration-dependent decrease (from 0.13 to 0.01 mL sec(-1) g(-1), or 7.4 to 0.56 times the blood perfusate flow rate) and was resolved as saturable and nonsaturable components, while data for retrograde were more scattered, varying from 2.8 to 1 times the blood perfusate flow rate. Efflux (coefficient = 0.0096 +/- 0.0024 and 0.0088 +/- 0.0062 mL sec(-1) g(-1), respectively) was relatively insensitive to concentration and flow direction. The same was observed for the removal capacity for metabolism and excretion (sequestration coefficient: for prograde, 0.0056 +/- 0.0017 mL sec(-1) g(-1); for retrograde, 0.0056 +/- 0.003 mL sec(-1) g(-1)). The decrease in the apparent partition coefficient (ratio of 4MUS concentration estimated in tissue to unbound plasma concentration) and the increase in relative throughput component with concentration further substantiate the claim on the presence of concentrative processes at the sinusoidal membrane.

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High-performance liquid chromatographic method for the direct determination of 4-methylumbelliferone and its glucuronide and sulfate conjugates

Cited by 20 publications

References 10 publications

The effects of 4-methylumbelliferone on hyaluronan synthesis, MMP2 activity, proliferation, and motility of human aortic smooth muscle cells

The effects of 4-methylumbelliferone on hyaluronan synthesis, MMP2 activity, proliferation, and motility of human aortic smooth muscle cells

Role of Breast Cancer Resistance Protein (Bcrp1/Abcg2) in the Extrusion of Glucuronide and Sulfate Conjugates from Enterocytes to Intestinal Lumen

Carrier-mediated entry of 4-methylumbelliferyl sulfate: Characterization by the multiple-indicator dilution technique in perfused rat liver

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