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Materials and Methods

Ethanol Tests. Identify acute alcohol ingestion. Identify ethanol, methanol, isopropanol, or acetone ingestion. Alcohols Method: Quantitative Gas Chromatography.

Methanol as a Marker of Alcohol Abuse

Ethyl Glucuronide and Ethyl Sulfate Tests. Preferred method for general screening of ethanol exposure in urine. Detect and document maternal use of ethanol. Chronic Alcohol Use Biomarkers. Carbohydrate-Deficient Transferrin Test. Phosphatidylethanol Test. Biomarker associated with ethanol consumption; may be helpful in monitoring alcohol abstinence Identify chronic heavy ethanol use for up to 28 days.

Nonspecific Biomarkers. Gamma-Glutamyl Transferase Test. Indirect marker associated with ethanol consumption. Aspartate Aminotransferase and Alanine Aminotransferase Tests. Biomarker associated with alcohol-induced liver damage Identify chronic heavy ethanol use.

References National Institutes of Health. National Institute on Alcohol Abuse and Alcoholism. Diagnostic and Statistical Manual of Mental Disorders, 5th ed. Arlington: Virginia: American Psychiatric Publishing, Marek E, Kraft WK. Ethanol pharmacokinetics in neonates and infants. Curr Ther Res Clin Exp. PubMed Kadehjian L. Specimens for Drugs-of-Abuse Testing, Ch.

Biomarkers of Heavy Drinking. Biomarkers of alcohol misuse: recent advances and future prospects. Prz Gastroenterol. Additional Resources. PubMed Ingall GB. Non-Invasive Assessment of Liver Fibrosis 58 min. Hemochromatosis Hemochromatosis Testing Algorithm.

Hepatocellular Carcinoma - HCC. Liver Disease Evaluation. Content Review:. August Ethanol, Serum or Plasma — Medical Carbohydrate Deficient Transferrin for Alcohol Use Pages Possible role of long chain fatty acid ethyl esters in organ injury and as short-term markers of ethanol intake in humans Diczfalusy, Margareta A. Methanol and congeners as markers of alcohol use and abuse Gilg, Thomas Pages The direct ethanol metabolite ethyl glucuronide: A specific marker of recent alcohol consumption Wurst, Friedrich Martin et al.

Phosphatidylethanol; clinical significance and biochemical basis Varga, Arthur et al. Proteins modified by direct and indirect ethanol metabolites, and their associated antibodies, as markers of alcohol intake Worrall, Simon Pages Salsolinol — An endogenous neurotoxin in the biology of alcoholism Musshoff, Frank Pages Show next xx. Read this book on SpringerLink. Strong hemolysis can also lead to false-positive results our unpublished data.

Conditions that need to be studied include positioning of the patient during blood collection, duration of congestion, in vivo and in vitro drug effects, use of EDTA- or heparin-plasma, and stability of whole-blood samples. This can be achieved by chromatographic e. Subsequently, the CDT isoforms are detected by immunological procedures. Regardless of which fractionation procedure and which immunoassay is used, a complete and stable Tf iron load is fundamental for reliable CDT analysis.

This point must be assessed when developing and launching a new CDT analytical method 11 Because of its high selectivity, IEF is used as the reference method for serum Tf isoform analysis. The Tf isoforms are separated in a gel containing a pH gradient according to their characteristic pIs. After electrophoresis, the Tf bands are visualized by immunofixation and staining of the CDT-anti-Tf complexes 3. Finally, the Tf band patterns can be evaluated by densitometry. It follows from this that the intensity of the main Tf fraction does not correlate with the amount of Tf focused in this band 3.

This complicates quantitative evaluation of this and the other Tf fractions. From the analytical point of view, this drawback can be overcome by use of ratios of the different CDT isoforms, e. Many IEF methods suggested for quantitative CDT analysis suffer from incomplete documentation of detection limits, recovery e. Because of its high selectivity, IEF can detect genetic Tf variants without in vitro neuraminidase treatment 10 The latter is done for complete removal of Tf isoform sialic acid residues and thus formation of only asialo-Tfs 10 13 Assuming complete Tf iron load and Tf sialic acid loss, only one band of asialo-Fe 2 -Tf would be detected in serum with a homozygous Tf type e.

IEF is suitable for visualization of Tf isoform patterns in the anion-exchange microcolumn eluates of commercial CDT tests. It is therefore an essential method for testing the reliability of the initial fractionation step used to separate CDT and non-CDT isoforms in current and upcoming CDT assays based on anion-exchange chromatography 11 In addition to IEF in flat gels, capillary electrophoresis 49 50 51 and capillary zone electrophoresis 15 52 53 have been proposed for analysis of Tf isoforms.

  • Microheterogeneity of Human Serum Tf;
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The main problem with these techniques is coating of the capillary surface to prevent protein adsorption and finding a coating-compatible, highly ultraviolet-transparent buffer Only a tradeoff between these criteria is achievable at present. Thus, it is questionable that Tagliaro et al. Recently, Crivellente et al. The improved sensitivity and separation were obtained mainly by the addition of diaminobutane to the running buffer and an increase in column length Nevertheless, the selectivity and sensitivity of capillary electrophoresis are still lower than IEF.

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When Tagliaro et al. Even with their improved method, Crivellente et al. Lectin affinity electrophoresis has been described for assessing Tf microheterogeneity in patients with alcoholic liver disease Whether these methods can gain wide acceptance for routine CDT analysis is still unclear.

Thus, genetic Tf variants cannot be detected by anion-exchange chromatography followed by immunoassay a procedure usually used by commercial CDT tests. Recently, Yoshikawa et al. These methods use the common CDT definition when analyzing asialo-, monosialo-, and disialo-Fe 2 -Tf. Test-specific cutoffs or borderlines for serum CDT concentration indicating chronic alcohol abuse.

The availability of CDT assays for routine laboratory diagnosis increased and accelerated the acceptance of CDT as one of the most specific markers of chronic alcohol abuse to date. Standardization of CDT analysis by an international CDT standard and quality-control material has not been attained [although it is possible to generate CDT enzymatically 55 56 58 ]. The lack of standardization of CDT analyses should not discourage the use of CDT, or encourage the use of less specific markers, e.

New and Upcoming Markers of Alcohol Consumption | Friedrich M. Wurst | Springer

Hemoglobin A 1C , which is widely used as an integrative long-term marker of blood glucose concentration, is far from being well standardized. Because of different analytical specificities and recoveries, normal ranges reference values or cutoffs for absolute and relative serum CDT are method-dependent. Therefore, the laboratory should report the CDT value, the cutoff value, and the method of analysis. Such information is common in forensic drug analysis; it makes the comparison of reports from different laboratories much easier, is needed in follow-up studies, and should also be provided for CDT analyses.

Relatively constant serum CDT concentrations have been found in healthy persons, in patients with non-alcohol-related liver diseases, or during abstinence 60 Individual reference ranges were valuable for follow-up during alcohol withdrawal treatment Absolute serum CDT concentrations from healthy women typically are higher than those of healthy men 9 31 63 64 65 Recently, de Feo et al.

The latter was also reported by van Pelt and Azimi Because females frequently have subclinical iron deficiency and thus increased Tf concentrations, this might explain their higher CDT concentrations.

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However, these findings are surprising because it is commonly accepted that CDT and Tf do not correlate see below. The reason for higher CDT concentrations in healthy women compared with healthy men remains unclear 63 64 , in spite of the findings of de Feo et al. No correlation has been found between CDT and the menstrual cycle, serum estradiol, serum iron, or contraception 65 Compared with premenopausal women, serum CDT was slightly increased Data pointing to a relationship between the amount and frequency of menstrual blood loss and CDT were recently presented by Leusink et al.

Increased serum CDT concentrations during pregnancy have been reported. Interestingly, de Jong and van Eijk 72 found a steady increase of tetrasialo-Tf and higher sialylated Tf isoforms and unaltered concentrations of sialic acid-deficient Tf isoforms in pregnant women during the second and third trimesters and also in women using contraceptives. In accordance with these observations, the CDT:Tf ratio should decrease and not increase as usually discussed in these women. This could be one reason for the diminished diagnostic sensitivity of serum CDT measurement in women compared with men see below.

Another underlying cause can be gender-dependent differences in the drinking patterns and in the vulnerability of the liver to alcohol intoxication, as discussed recently Except for two recent studies 67 68 , no significant correlation between serum CDT and total Tf has been found 9 14 74 Determining this ratio requires IEF, which is sometimes laborious and requires experienced personnel. This might be one reason why the Tf-index has not gained much acceptance. Automated IEF, such as that described by Hackler et al.

Whether this ratio increases the diagnostic accuracy compared with absolute CDT concentrations and in each situation is controversial 7 19 31 44 48 77 78 79 80 Contradictory results were found in a single study using CDT:Tf ratios 67 , which demonstrated improved diagnostic specificity in patients with iron-deficiency anemia but no decrease in false-negative results in patients with alcohol abuse.

However, low total serum Tf concentrations e. Thus, Lieber 48 found in some liver disease patients increased CDT:Tf ratios that were attributable to decreased serum Tf concentrations. When borderlines of 2. At the same time, the absolute CDT concentration does not exceed the upper limit of the borderline. Knowing this is important for interpretation of CDT values in longitudinal studies or for control of relapse drinking. Our investigations revealed that measuring and interpreting absolute and relative CDT concentrations as well as the total serum Tf concentration improves the diagnostic efficiency of CDT as a marker of chronic alcohol abuse unpublished data.

It might be interesting to reevaluate the data from the many clinical studies, taking into consideration absolute serum CDT concentrations, CDT:Tf ratios, and total serum Tf concentration. Reviews of the literature up to 82 and between and November 83 have been published. Since then, several studies on these subjects have appeared. Numerous clinical trials have been published regarding the diagnostic specificity and sensitivity of serum CDT as a marker of chronic alcohol abuse.

Several reviews have been published 9 31 82 94 Since then, many new reports have appeared in journals listing impact factors. Most of these reports have already been discussed here. It is not the aim of this review to compare the diagnostic criteria e. For calculating parameters of diagnostic efficiency, reliable data regarding individual alcohol consumption are a prerequisite.

Clinical background or personal reviews are customarily used for this purpose. However, if both were faultless, we would not need laboratory markers of chronic alcohol abuse. Because underestimating alcohol consumption is common, false true positives or false false negatives will occur and markedly affect the criteria of diagnostic efficiency of CDT obtained in a specific clinical study.

This should be considered when interpreting parameters of diagnostic efficiency. The main causes for false-positive results in CDT analysis have been described by Stibler and co-workers 9 Diagnostic specificities of CDT obtained in several different clinical studies have been published 9 31 82 83 Clinical conditions, regardless of the number of observations, that caused false positives when serum CDT was used as a marker of chronic alcohol abuse. Differences in the drinking patterns during an appropriate period before blood sampling long-term drinking pattern and within the individual days of this period short-term drinking patterns must be assessed.

The results of a 3-week drinking experiment showed that chronic consumption of small amounts of alcohol affected serum CDT concentrations [confirmed by Whitfield et al. Because of ethical considerations, long-time drinking experiments are difficult to perform. However, it is known that ethanol blood concentrations are affected not only by the amount of alcohol consumed, but also by the amount per time period and by the body mass distribution volume : the shorter the period in which a gram of alcohol is consumed and the lower the body mass, the higher the peak blood alcohol concentration.

Thus, one might hypothesize that chronic short-term consumption of high amounts of alcohol by a lean person causes high peak blood alcohol concentrations and substantial liver injury, and thus affects serum CDT concentrations or the dose—response curve.

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A relationship between CDT insulin sensitivity and hypertension in men of a very specific patient sample were discussed by Fagerberg et al. Conditions reported to affect diagnostic sensitivity of serum CDT concentration as a marker of chronic alcohol abuse.

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The main concern about CDT as a marker of chronic alcohol abuse is its relatively low diagnostic sensitivity. Subsequent studies, reporting distinctly lower diagnostic sensitivities, could not meet these expectations [reviewed in Refs. Thus, diagnostic specificities of The shortcomings of this study were discussed by Allen and Sillanaukee and Arndt et al. CDT analytical methods with different analytical specificities and recoveries decrease the comparability of CDT values and statistical parameters of the diagnostic efficiency of CDT obtained in different studies.

A unique definition for and standardization of CDT are needed. CDT is not a screening tool for detection of increased alcohol consumption and it was originally not aimed as such. Skip to main content. Research Article Reviews. Torsten Arndt. Published January Abstract Background: Carbohydrate-deficient transferrin CDT is used for diagnosis of chronic alcohol abuse. Microheterogeneity of Human Serum Tf Tf, the most important iron-transport protein, is synthesized mainly in hepatocytes and consists of three substructural domains: a single polypeptide chain, two independent metal ion-binding sites one within the N-terminal and the other within the C-terminal domain , and two N-linked complex glycan chains Fig.

Figure 1. Figure 2. Structure of Human Serum CDT Serum samples from alcoholics showed decreased sialic acid content 20 and normal Tf isoform patterns after treatment with neuraminidase complete removal of sialic acid residues from the Tf N-glycan chains Pathomechansisms of Ethanol-induced CDT Increase The pathomechanisms for the increase in CDT isoforms during chronic alcohol abuse are not completely understood at present.

Preanalysis Several preanalytical conditions have been found to affect serum CDT concentrations. View this table: View inline View popup. Table 1. Medical Interpretation Postanalysis normal ranges for serum cdt Because of different analytical specificities and recoveries, normal ranges reference values or cutoffs for absolute and relative serum CDT are method-dependent. Table 2. Table 3. Acknowledgments I thank Lloyd Allen Jones for stylistic emendations. Isoelectric focusing and electrophoresis of the CSF proteins in tremor of different origins.

J Neurol Sci ; 30 : The biology of transferrin. Clin Chim Acta ; : 1 Effect of separation conditions on automated isoelectric focusing of carbohydrate-deficient transferrin and other human isotransferrins using the PhastSystem. Anal Biochem ; : Purification of isotransferrins by concanavalin A Sepharose chromatography and preparative isoelectric focusing.

Human serum sialo transferrins in diseases. Clin Chim Acta ; : The analysis of human serum transferrins with the PhastSystem: quantitation of microheterogeneity. Electrophoresis ; 13 : Transferrin isoform distribution: gender and alcohol consumption. Alcohol Clin Exp Res ; 21 : Human transferrin polymorphism. Hum Hered ; 37 : 65 Stibler H. Carbohydrate-deficient transferrin in serum: a new marker of potentially harmful alcohol consumption reviewed. Clin Chem ; 37 : Bean P , Peter JB. Allelic D variants of transferrin in evaluation of alcohol abuse: differential diagnosis by isoelectric focusing-immunoblotting-laser densitometry.

Clin Chem ; 40 : Clin Chem ; 46 :