Abstract:
BACKGROUND:Cleavage of conjugates is an important step in toxicological analysis, especially of urine samples. The aim of this study was to combine the advantages and to reduce the disadvantages of acid hydrolysis and conventional enzymatic hydrolysis procedures. METHODS:beta-Glucuronidase (GRD; EC 3.2.1.31) and arylsulfatase (ARS; EC 3.1.6.1) were purified and coimmobilized on an agarose gel matrix and packed into columns. RESULTS:In columns packed with GRD and ARS, the test conjugates 4-nitrophenyl glucuronide and 4-nitrophenyl sulfate added into urine could be completely cleaved within 25 min. Even the relatively stable morphine conjugates could be completely hydrolyzed within 60 min in authentic urine samples. Therefore, an incubation time of 1 h is recommended. Enzyme inhibition by matrix or by rather high concentrations of acetaminophen conjugates was tested and found to be up to 50%. However, a large excess of GRD and ARS was used. The immobilizate columns could be reused for at least 70 incubations and had a storage stability of at least 12 weeks. Carryover of analytes in reused columns could be avoided by rinsing with 200 mL/L methanol in acetate buffer. Thus, five drugs known to be contaminants added in very high concentrations into urine could be completely removed from the columns. A study on the applicability in systematic toxicological analysis showed that 120 different drugs and/or their metabolites could be detected in 35 different authentic urine samples. CONCLUSIONS:Use of immobilized and column-packed GRD and ARS is an efficient alternative for the cleavage of urinary conjugates in clinical toxicology.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Toennes SW,Maurer HHsubject
Has Abstractpub_date
1999-12-01 00:00:00pages
2173-82issue
12eissn
0009-9147issn
1530-8561journal_volume
45pub_type
杂志文章abstract:BACKGROUND:The American Diabetes Association task force on standardization of insulin assays in 1996 showed wide variation in assay bias. Newer assays are specific for insulin, with several now available on automated immunoassay analyzers. METHODS:In 2004, we compared 11 commercially available insulin assays by analyz...
journal_title:Clinical chemistry
pub_type: 杂志文章,多中心研究
doi:10.1373/clinchem.2006.077784
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journal_title:Clinical chemistry
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pub_type: 杂志文章
doi:10.1373/clinchem.2006.077859
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
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journal_title:Clinical chemistry
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doi:
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doi:
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journal_title:Clinical chemistry
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1998-01-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1979-06-01 00:00:00
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journal_title:Clinical chemistry
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doi:
更新日期:1984-12-01 00:00:00
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journal_title:Clinical chemistry
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1982-05-01 00:00:00
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更新日期:2014-10-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2005.062414
更新日期:2006-05-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1994-09-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:2002-06-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1992-01-01 00:00:00
abstract::A new method for quantifying specific amino acids in small volumes of plasma and whole blood has been developed. Based on isotope-dilution tandem mass spectrometry, the method takes only a few minutes to perform and requires minimal sample preparation. The accurate assay of both phenylalanine and tyrosine in dried blo...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1993-01-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1995-03-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1975-07-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1998-01-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1990-09-01 00:00:00
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pub_type: 杂志文章
doi:
更新日期:1997-08-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
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更新日期:2019-01-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1980-10-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1989-10-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1985-04-01 00:00:00
abstract::Of 98 patients' specimens examined for alkaline phosphatase (EC 3.1.3.1) isoenzymes by electrophoresis on cellulose acetate membrane after incubation with neuraminidase, 50 showed only a single liver or bone isoenzyme staining band; in 15 of these, the tissue origin of the fraction could not be accurately identified f...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1985-07-01 00:00:00
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journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1989-06-01 00:00:00