Abstract：
BACKGROUND:Simple, reproducible assays are needed for the quantification of sphingolipids, ceramide (Cer), and sphingoid bases. We developed an HPLC method for simultaneous quantification of total plasma concentrations of Cer, glucosylceramide (GlcCer), and ceramide trihexoside (CTH). METHODS:After addition of sphinganine as internal calibrator, we extracted lipids from 50 microL plasma. We deacylated Cer and glycosphingolipids by use of microwave-assisted hydrolysis in methanolic NaOH, followed by derivatization of the liberated amino-group with o-phthaldialdehyde. We separated the derivatized sphingoid bases and lysoglycosphingolipids by HPLC on a C18 reversed-phase column with a methanol/water mobile phase (88:12, vol/vol) and quantified them by use of a fluorescence detector at lambda(ex) 340 nm and lambda(em) 435 nm. RESULTS:Optimal conditions in the Solids/Moisture System SAM-155 microwave oven (CEM Corp.) for the complete deacylation of Cer and neutral glycosphingolipids without decomposition were 60 min at 85% power, fan setting 7. Intra- and interassay CVs were <4% and <14%, respectively, and recovery rates were 87%-113%. The limit of quantification was 2 pmol (0.1 pmol on column), and the method was linear over the interval of 2-200 microL plasma. In samples from 40 healthy individuals, mean (SD) concentrations were 9.0 (2.3) micromol/L for Cer, 6.3 (1.9) micromol/L for GlcCer, and 1.7 (0.5) micromol/L for CTH. Plasma concentrations of GlcCer were higher in Gaucher disease patient samples and of CTH in Fabry disease patient samples. CONCLUSIONS:HPLC enables quantification of total Cer, GlcCer, and CTH in plasma and is useful for the follow-up of patients on therapy for Gaucher or Fabry disease.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Groener JE,Poorthuis BJ,Kuiper S,Helmond MT,Hollak CE,Aerts JMdoi
10.1373/clinchem.2006.079012subject
Has Abstractpub_date
2007-04-01 00:00:00pages
742-7issue
4eissn
0009-9147issn
1530-8561pii
clinchem.2006.079012journal_volume
53pub_type
杂志文章abstract：BACKGROUND:The next-generation, high-sensitivity cardiac troponin assays can measure quantifiable concentrations of cTn in a majority of individuals, but there are few studies assessing these assays for risk stratification. The present study was undertaken to determine if a research hs-cTnI assay can be useful for pred...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2009.127241
更新日期：2009-10-01 00:00:00
abstract：:Each of 65 laboratories analyzed 10 whole-blood samples for erythrocyte protoporphyrin by one or more of several analytical procedures. These procedures were of two types: (a) extraction of protoporphyrin from the erythrocytes into ethyl acetate/acetic acid, re-extraction into hydrochloric acid, and fluorometric measu...
journal_title：Clinical chemistry
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doi：
更新日期：1978-12-01 00:00:00
abstract：BACKGROUND:In this study we demonstrate the technical application of flow cytometry and cell sorting combined with gene-rearrangement clonality profiling to detect and confirm minimal disease in 2 leukemia and 2 lymphoma cases. METHODS:Specimens with low percentages (0.05%-5%) of abnormal lymphoid populations were ide...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2005.061259
更新日期：2006-03-01 00:00:00
abstract：:We evaluated the EMIT (enzyme-multiplied immuno technique) and FPIA (fluorescence polarization immunoassay) urine screening systems for detection of benzodiazepine intake. Healthy male volunteers were given single oral therapeutic doses of alprazolam (2 mg), chlordiazepoxide (25 mg), flunitrazepam (1 mg), lorazepam (3...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1992-02-01 00:00:00
abstract：:This solid-phase competitive europium immunoassay of progesterone in plasma relies on antigen labeling. With this new approach, time-resolved fluoroimmunoassay can attain sensitivity and precision similar to that of conventional radioimmunoassay. The europium-labeling involves coupling diethylenetriaminepentaacetic ac...
journal_title：Clinical chemistry
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doi：
更新日期：1988-03-01 00:00:00
abstract：:In this rapid, simple enzyme immunoassay for carcinoembryonic antigen (CEA) in serum the "sandwich" system is used. The CEA is first extracted from the serum into acetate buffer by heating at 70 degrees C, then incubated in a polyvinyl microtitration plate previously coated with anti-CEA. The bound CEA is then reacted...
journal_title：Clinical chemistry
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更新日期：1983-12-01 00:00:00
abstract：BACKGROUND:The determination of cellular beta-galactocerebrosidase activity is an established procedure to diagnose Krabbe disease and monitor the efficacy of gene/stem cell-based therapeutic approaches aimed at restoring defective enzymatic activity in patients or disease models. Current biochemical assays for beta-ga...
journal_title：Clinical chemistry
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doi：10.1373/clinchem.2008.115873
更新日期：2009-03-01 00:00:00
abstract：:Hepatitis A virus (HAV) infection occurs worldwide and is an important cause of acute viral hepatitis in the US. In this review, I cover the epidemiology, course of infection, clinical manifestations, serological responses, and prevention of this infection. Although most patients completely recover from this disease, ...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：
更新日期：1997-08-01 00:00:00
abstract：:The isoenzyme pattern of N-acetyl-beta-glucosaminidase (NAG) in serum and urine was studied in two groups of patients with diabetes mellitus and in 30 control subjects. Total NAG activity was significantly (P less than 0.001) increased in the serum and urine of the 20 diabetics with vascular complications, but was ins...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1988-12-01 00:00:00
abstract：:A micro version of a nuclear binding assay to assess the biological activity of receptors for steroid hormones was developed for application to small (needle) biopsies of human tumors for the purpose of predicting responses to steroid therapy. This easier assay requires 10-fold less tissue than the original nuclear bi...
journal_title：Clinical chemistry
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doi：
更新日期：1988-02-01 00:00:00
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journal_title：Clinical chemistry
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doi：
更新日期：1996-08-01 00:00:00
abstract：:We describe a high-performance liquid chromatographic method for the quantitative determination of gentamicin in serum. The antibiotic was separated from serum by passage through a silicic acid column, derivatized with o-phthalaldehyde, and eluted with ethanol. The derivatized gentamicin was then separated into all th...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1977-12-01 00:00:00
abstract：:In the disposable rotor of the SAM microcentrifugal analyzer, various lyophilized reagents are predistributed in 24 33-microL cuvets, for determination of multiple analytes in one specimen (e.g., for patient profiles). We evaluated a prototype of this system, which can be used at 25, 30, or 37 degrees C; absorbance re...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1985-09-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1982-03-01 00:00:00
abstract：BACKGROUND:There is extended urinary excretion of Δ(9)-tetrahydrocannabinol (THC), 11-hydroxy-THC (11-OH-THC), and 11-nor-9-carboxy-THC (THCCOOH) in abstinent frequent cannabis smokers. We characterized THC, 11-OH-THC, THCCOOH, cannabidiol, cannabinol, THC-glucuronide, and THCCOOH-glucuronide disposition in urine of fr...
journal_title：Clinical chemistry
pub_type： 临床试验,杂志文章
doi：10.1373/clinchem.2013.214106
更新日期：2014-02-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1991-12-01 00:00:00
abstract：:Serum concentrations of apolipoprotein (apo) B and apo A-I were measured from 1985-1996 in a Swedish population sample of 83 112 males and 64 464 females, ages <20 to >80 years, using an automated immunoturbidimetric method calibrated against fresh pools of human serum and commercial calibrators. All values were recal...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1998-08-01 00:00:00
abstract：BACKGROUND:Quantification of 5-hydroxyindole-3-acetic acid (5-HIAA) in urine is useful in diagnosing and monitoring of patients with carcinoid tumors and in the study of serotonin (5-hydroxytryptamine) metabolism in various disorders. We describe an automated method that incorporates on-line solid-phase extraction (SPE...
journal_title：Clinical chemistry
pub_type： 杂志文章
doi：10.1373/clinchem.2005.050062
更新日期：2005-09-01 00:00:00
abstract：:Various laboratories have reported endogenous digoxin-like immunoreactive factor(s) (DLIF) in blood from patients in renal failure or liver failure, from newborn infants, and from third-trimester pregnant women. Similar immunoreactivity has been detected in amniotic fluid, in cord blood, and in urine and serum from no...
journal_title：Clinical chemistry
pub_type： 杂志文章,评审
doi：
更新日期：1985-09-01 00:00:00
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journal_title：Clinical chemistry
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doi：
更新日期：1976-12-01 00:00:00
abstract：BACKGROUND:The Elecsys beta-CrossLaps serum assay measures type I collagen degradation fragments (beta-CTx) that contain the beta-isomerized octapeptide EKAHD-beta-GGR. We investigated the analytical performance of the assay and changes in beta-CrossLaps in patients with metabolic bone diseases. METHODS:The electroche...
journal_title：Clinical chemistry
pub_type： 临床试验,杂志文章
doi：
更新日期：2001-08-01 00:00:00
abstract：:To see if saliva is a valid substitute for plasma in assay of progesterone even when concentrations of hormone and binding proteins are fluctuating, we determined the concentrations of total and free progesterone in plasma and salivary progesterone in specimens from 36 women volunteers during the course of pregnancy a...
journal_title：Clinical chemistry
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doi：
更新日期：1989-01-01 00:00:00
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journal_title：Clinical chemistry
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doi：
更新日期：2000-01-01 00:00:00
abstract：BACKGROUND:The majority of patients with chronic kidney disease are diagnosed and monitored in primary care. Glomerular filtration rate (GFR) is a key marker of renal function, but direct measurement is invasive; in routine practice, equations are used for estimated GFR (eGFR) from serum creatinine. We systematically a...
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pub_type： 杂志文章,meta分析
doi：10.1373/clinchem.2017.276683
更新日期：2018-03-01 00:00:00
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journal_title：Clinical chemistry
pub_type： 杂志文章
doi：
更新日期：1982-04-01 00:00:00
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journal_title：Clinical chemistry
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doi：
更新日期：1987-04-01 00:00:00
abstract：:In routine analysis for immunoglobulin light chains in pediatric diagnostics, the age-related reference intervals for serum kappa (kappa) and lambda (lambda) light chains were evaluated in 1543 healthy subjects (newborns to age 16 years, including 168 premature infants). Light-chain analysis was performed by rate neph...
journal_title：Clinical chemistry
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doi：
更新日期：1992-04-01 00:00:00
abstract：:We evaluated the enzymic mechanism by which 3-keto butylidene-beta-2-chloro-4-nitrophenyl maltopentaoside (3KB-G5-CNP) serves as a substrate for serum pancreatic (p-) and salivary (s-) amylases. In aliquots of the reaction mixture, three kinds of beta-2-chloro-4-nitrophenyl oligosaccharides (glucose, maltoside, and ma...
journal_title：Clinical chemistry
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doi：
更新日期：1991-08-01 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：1981-02-01 00:00:00
