Studies on the Determination of Bile Pigments

1964 ◽  
Vol 10 (2) ◽  
pp. 95-102 ◽  
Author(s):  
G W Stevenson ◽  
S L Jacobs ◽  
R J Henry

Abstract The absorbance at 450 mµ of serum diluted with acidified ethylene glycol is used as a measure of the serum's total bilirubin content in a single-tube, single-extraction method for rapid determination of free and total bilirubin in 0.1 ml. of serum. With the addition of chloroform, free bilirubin is extracted into the chloroform but conjugated bilirubin remains in the ethylene glycol. Free bilirubin is determined from the absorbance at 450 mµ in the chloroform layer. Conjugated bilirubin is calculated from the difference. Absorbance readings at 520 mµ compensate for the absorbance at 450 mµ, due to hemoglobin in the serum. Comparison with the Malloy-Evelyn procedure showed equivalent levels of total but higher levels of conjugated bilirubin.

1973 ◽  
Vol 19 (12) ◽  
pp. 1366-1369 ◽  
Author(s):  
Gerhard Ertingshausen ◽  
Diane L Fabiny Byrd ◽  
Thomas O Tiffany ◽  
Sandra J Casey

Abstract We report a rapid single-reagent method for determining total bilirubin with the " CentrifiChem Analyzer" (Union Carbide Corp.). The reagent is the stable 1,5-disulfonate of diazosulfanilic acid, and ethylene glycol is used as an accelerator. The reaction requires less than 5 min. Only 25 µl of serum is needed. We present precision data (x = 2.62 mg/dl, SD = .082 mg/dl, CV = 3.13%) and results (r = .998) as compared with those obtained by an automated Jendrassik—Grof method.


1964 ◽  
Vol 10 (5) ◽  
pp. 433-439 ◽  
Author(s):  
S L Jacobs ◽  
R J Henry ◽  
M Segalove

Abstract Tests for total bilirubin on 10 icteric sera by 5 diazo procedures and a spectrophotometric method showed fair agreement with most samples but an occasional grossly discrepant result. Results for free and conjugated bilirubin by diazo methods, acetone precipitation, and extraction of serum with ethyl acetate and chloroform were widely discrepant. Problems in documentation of accuracy of such procedures are discussed. It is concluded that unavailability of reference standards of bilirubin conjugates poses a serious obstacle to solution of the analytical problems.


1996 ◽  
Vol 79 (4) ◽  
pp. 907-916 ◽  
Author(s):  
Joanna M Lynch ◽  
David M Barbano ◽  
J Richard Fleming

Abstract A modified Mojonnier ether extraction method for determination of the fat content of cream was developed based on the method for milk (AOAC Official Method 989.05). The cream Babcock method (AOAC Official Method 920.111 B-C) was modified to harmonize with the milk Babcock method (AOAC Official Method 989.04) and to clarify procedural details. Using the AOAC collaborative study format, 10 laboratories tested 9 pairs of blind duplicate heat-treated cream samples with a fat range of 30-45% using both methods. The statistical performance (invalid and outlier data removed) was as follows: mean % fat = 37.932, sr = 0.125, sR = 0.151, RSDr = 0.330, RSDR = 0.398, r = 0.354, and R = 0.427 for the ether extraction method. For the Babcock method, mean % fat = 38.209, sr = 0.209, SR = 0.272, RSDr = 0.548, RSDR = 0.712, r = 0.592, and R = 0.769. Average test results for fat from the Babcock method were 0.277% (absolute fat) greater than for the Mojonnier ether extraction method. The difference between methods, as a percentage of the average fat content of the samples, was 0.73%. This agrees with differences observed between the 2 methods for milk when 10 to 17 laboratories tested 7 milk samples in blind duplicate at bimonthly intervals over a 4-year period (average difference 0.029% fat, 0.78% as a percentage of average fat content). The Mojonnier ether extraction and Babcock methods for fat in cream have been adopted by AOAC INTERNATIONAL. The new Babcock method replaced the AOAC Official Method 920.111 B-C.


1979 ◽  
Vol 62 (3) ◽  
pp. 662-670
Author(s):  
Hussein S Ragheb

Abstract Seventeen laboratories evaluated the pyridine extraction method and neomycin-sensitized agar for the determination of zinc and MD bacitracin in swine and broiler rations at 10 and 100 g/ton. The method was also applied to the analysis of 2 premixes labeled 50 g/lb (MD bacitracin) and 40 g/lb (zinc bacitracin). Bacitracin activity was determined on each of 2 days with 2 dilutions on each day. No significant difference was found between dilutions within a day or between days for each sample. The type of bacitracin or type of feed did not significantly affect results. The difference in results between MD and zinc bacitracin in premixes approached significance. The large coefficients of variation for premixes (ca 13%) and complete feeds (ca 15–30%) indicate operational problems. The main difficulty was evaporation of pyridine. Some laboratories were not able to evaporate it completely, whereas others lost bacitracin activity, probably due to high temperature of drying. The pyridine extraction method as in 42.200 and 42.204 should be discontinued.


1989 ◽  
Vol 35 (8) ◽  
pp. 1760-1762 ◽  
Author(s):  
D W Franquemont ◽  
J L Sutphen ◽  
D A Herold ◽  
D E Bruns

Abstract We describe the cases of four patients who were taking sulfasalazine for inflammatory bowel disease, whose conjugated bilirubin concentrations in serum exceeded their corresponding total bilirubin concentrations as measured with a multilayer film analyzer, the Ektachem 400. Sulfasalazine added to pooled human serum at therapeutic concentrations increased the apparent conjugated bilirubin, as measured with the Ektachem, in a linear and dose-related fashion. Measured unconjugated bilirubin was simultaneously decreased to values less than -3 mg/L. The same interference occurred on the Ektachem 700, but an algorithm prevented the instrument from reporting the results. The major metabolites of sulfasalazine in blood did not interfere with analysis for those fractions of bilirubin. Sulfasalazine's strong absorbance at 400 nm explains its interference with determination of conjugated bilirubin in this instrument.


1993 ◽  
Vol 27 (12) ◽  
pp. 1429-1433 ◽  
Author(s):  
Robert J. Kandrotas ◽  
Peter Gal ◽  
Jean B. Douglas ◽  
James B. Groce

OBJECTIVE: To compare heparin dosage adjustment using only activated partial thromboplastin time (APTT) with a method using non-steady-state heparin concentrations (HCs) to rapidly achieve and maintain an APTT ratio greater than or equal to 1.5 times baseline throughout the first 24 hours of therapy. DESIGN: Randomized, blind, parallel comparison of an empiric dosing method based only on APTT with a dosing method based on the calculation of heparin clearance using non-steady-state HCs. SETTING: A private community teaching hospital. The patient, physician, nurses, and investigators were blinded to the dosing method. Only the clinical staff pharmacist, who received the consult and made all dosage adjustments, was not blinded. PATIENTS: All patients requiring heparin for the treatment of thromboembolic disease were evaluated for potential inclusion in the study. Patients were enrolled in the study if they had a clinical diagnosis of deep venous thrombosis confirmed by objective means such as venography or ultrasonography. Patients were excluded if they had active bleeding, platelet dysfunction, thrombocytopenia, severe hepatic disease (total bilirubin >25.7 μmol/L), renal disease, or evidence of stroke. Patients were also excluded if they were receiving heparin prior to enrollment. MAIN OUTCOME MEASURE: Maintenance of an APTT ratio greater than or equal to 1.5 times baseline throughout the first 24 hours of heparin therapy. RESULTS: Thirty-four patients were enrolled in the study; 17 in each group. The groups were not significantly different with regard to gender, age, baseline APTT, or mean loading dose (p>0.5). Mean initial infusion rates for the control and HC groups were 1042 ± 194 and 1071 ± 143 units/h, respectively (p>0.5). After the first rate adjustment at 4 hours, the difference achieved significance at 1032 ± 232 and 1367 ± 317 units/h for the control and HC groups, respectively (p<0.01). At 12 hours, 18.8 percent of the patients in the control group were subtherapeutic; by 24 hours, 33.3 percent were subtherapeutic. No patients became subtherapeutic in the HCs group during the first 24 hours. CONCLUSIONS: This study demonstrates that, in contrast to standard heparin dosing methods, the use of non-steady-state HCs allows patients with deep venous thrombosis to rapidly achieve and maintain therapeutic APTT ratios throughout the critical first 24 hours of therapy.


1980 ◽  
Vol 26 (1) ◽  
pp. 22-25 ◽  
Author(s):  
B C Shull ◽  
H Lees ◽  
P K Li

Abstract Oxyhemoglobin is the species of hemoglobin in erythrocyte hemolysates that inhibits the diazo reaction. Ferric hemoglobin derivatives and species with relatively low molecular mass do not interfere. Conversion of oxyhemoglobin to acid hematin under assay reaction conditions is associated with rapid destruction of bilirubin, which accounts for the diazo reaction error. The most probable mechanism for this destruction of bilirubin is an oxidative reaction involving H2O2, formed in the oxidation of hemoglobin, and acid hematin acting as a pseudoperoxidase. We could find no evidence for other mechanisms of interference such as spectral error or azobilirubin destruction. Addition of potassium iodide, 4.0 mmol/L final concentration in the reaction mixture, eliminates interference from hemoglobin added to give concentrations as great as 10 g/L. It also eliminated the effects of hemolysis in the method of Ertingshausen et al. (Clin. Chem. 19: 1366, 1973), in which ethylene glycol is used as the accelerator.


1999 ◽  
Vol 82 (4) ◽  
pp. 792-798
Author(s):  
Richard E Mauldin ◽  
John J Johnston ◽  
Craig A Riekena

Abstract A liquid extraction method is described that permits rapid determination of cholecalciferol (D3) in rodenticidal grain baits. Purified D3 was incubated for various time periods to produce pre-D3. Response ratios (concentration/detector response) for various concentrations of pre-D3 and D3 in solutions permitted generation of a correction factor for direct quantitation of pre-D3 in solutions with a pure D3 standard. The method has equal precision and accuracy, yet is simpler and less time consuming and requires less solvents than widely accepted methods for extracting D3 from grain baits. Recoveries from control oat baits fortified at 0.05 and 0.75 wt% were 100.9 and 98%, respectively. A standard curve for concentrations ranging from 6.4 to 204 μg/mL had an r2 of 0.9999 and an intercept of zero and was linear and proportional. The method limit of detection was 2.0 × 10−4 wt % D3.


2013 ◽  
Vol 448-453 ◽  
pp. 359-362
Author(s):  
Mo Jie Sun ◽  
Ling Zhang ◽  
Ruo Kun Jia

Chloral alkali at room temperature Can be quickly converted into chloroform completely, Application of this principle, By Purge and Trap GC/MS method , Determination of the alkali content of chloroform in water before and after the difference, Inverse to get the water content of chloral. NaOH were added to different volumetric flask. It is flask with standard solutions in different concentrations. Aside a certain amount of determination, error is larger. NaOH was injected directly into the injector. This was the method. It completely transformed in Purge and Trap, greatly reduces human error. The linear range is 0.5-20ug/L, the minimum detection limit can reach 0.05ug/L, the relative standard deviation is less than 2.3%, the average recovery was 97.5%. This method is simple and quick,the results are accurate and reliable, which is able to meet the drinking water source in the analytical needs of chloral.


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