PHOTOMETRIC STUDIES ON THE DIAZO REACTION FOR SERUM BILIRUBIN

1952 ◽  
Vol 30 (6) ◽  
pp. 552-560
Author(s):  
F. D. White ◽  
Dorothea Duncan

Photometric studies on the Malloy-Evelyn procedure for the determination of serum bilirubin have shown that the azobilirubin color intensity obtained when the diazo reagent acts for 30 min. upon a known amount of bilirubin added to serum in the form of the sodium salt is less than when the reaction takes place with the same amount of bilirubin in chloroform-alcohol solution. This suggests that the usual calibration curve prepared from bilirubin in chloroform-alcohol solution, when used as standard for serum bilirubin determinations, may give values which are about 10% less than the true bilirubin content of the serum. It has also been shown that with the 1 : 10 dilution of the Malloy-Evelyn procedure the azobilirubin from icteric sera does not obey Beer’s law beyond a serum bilirubin content of 15 mgm. per 100 ml. Evidence is submitted that the rate of azobilirubin color formation can be markedly accelerated by increasing the strength of the diazo reagent, and a new reagent is proposed by the use of which the time of color development can be reduced from 30 min. to 5 min.

1974 ◽  
Vol 20 (10) ◽  
pp. 1362-1363 ◽  
Author(s):  
V Chromý ◽  
J Fischer ◽  
V Kulhánek

Abstract We studied the optimal composition of EDTA-chelated biuret reagent for the determination of protein in serum. A reagent containing 18 mmol of EDTA, 15 mmol of Cu2+, and 1 mol of NaOH per liter exhibits a very low blank but about the same sensitivity as tartrate-chelated reagents. Maximum color intensity is attained within 25 min at room temperature, and adheres to Beer's law up to 120 g of protein per liter.


1985 ◽  
Vol 33 (7) ◽  
pp. 729-732 ◽  
Author(s):  
R A Clark ◽  
G L Krueger

The triammonium salt of aurin tricarboxylic acid, commonly referred to as aluminon, forms a dye that has been used for the colorimetric determination of Al(III) species. We have reviewed the pertinent literature on the reaction of aluminon with respect to the metallic species that form colored aluminon complexes. The effects of experimental variables, such as time, temperature, and pH, upon the color development of the aluminon complex are also presented. Organic and inorganic species, particularly Be(II) and Fe(III), which can affect color formation, are described. The use of aluminon as a histochemical staining agent for the detection of aluminum requires verification by atomic absorption spectrophotometric analysis or other quantitative techniques.


1990 ◽  
Vol 38 (1) ◽  
pp. 87-94 ◽  
Author(s):  
M W Wessendorf ◽  
S J Tallaksen-Greene ◽  
R M Wohlhueter

7-Amino-4-methylcoumarin-3-acetic acid (AMCA) has been found to be a useful fluorophore for immunofluorescence. The present study describes a spectrophotometric method for determining the ratio of moles AMCA to moles protein (or the f/p ratio) in an AMCA-conjugated IgG. The concentration of a substance absorbing light can be determined spectrophotometrically using Beer's Law: Absorbance = Concentration x Extinction coefficient. From Beer's law, one can derive the following formula for determining the f/p ratio of AMCA-IgG conjugates: f/p = (epsilon 280IgG).A350 - (epsilon 350IgG).A280/(epsilon 350AMCA).A280 - (epsilon 280AMCA).A350 where A is the optical density of the conjugate at the given wavelength and epsilon is the extinction coefficient of a substance at the wavelength specified. Using conjugates of model proteins, it was found that the extinction coefficients of the AMCA moiety of AMCA-conjugated protein were 1.90 x 10(4) at 350 nm and 8.29 x 10(3) at 280 nm. Similarly, it was found that the extinction coefficients of swine IgG were 1.56 x 10(3) at 350 nm and 1.26 x 10(5) at 280 nm. Thus, for AMCA-conjugated swine IgG: f/p = (1.26 x 10(5)).A350 - (1.56 x 10(3)).A280/(1.47 x 10(4)).A280 - (6.42 x 10(3)).A350 [corrected]. Based on this formula, the f/p ratios of some AMCA-IgG conjugates useful for immunohistochemistry have been found to range between 6 and 24.


2010 ◽  
Vol 7 (2) ◽  
pp. 1001-1005
Author(s):  
Baghdad Science Journal

A rapid high sensitive and inexpensive economic method has been developed for the Determination of phenoxazine by using molecular spectrophotometry. The method is based on the oxidation of phenoxazine by potassium (meta)periodate in acidic medium. The oxidation conditions were selected to enhance the sensitivity and the stability of the pink colored species which shows an absorption maximum at 530 nm. The Beer’s law was obeyed for phenoxazine concentration range from 1 to 6 µg mL-1 with 0.003 µg mL-1 detection limit and provided variation coefficients between 0.4 to 1.7 %. This method was successfully applied for the determination of phenoxazine in aqueous samples


1949 ◽  
Vol 27e (3) ◽  
pp. 159-163 ◽  
Author(s):  
J. M. R. Beveridge ◽  
S. E. Johnson

A simple accurate method for the determination of phospholipid phosphorus is described. Conditions for the digestion of phospholipid have been developed permitting the utilization of a colorimetric procedure for inorganic phosphorus that is characterized by excellent color stability and strict adherence to Beer's law over a range of 0 to 65 μgm. of phosphorus.


1975 ◽  
Vol 58 (6) ◽  
pp. 1232-1234
Author(s):  
Jois R Rangaswamy ◽  
Yadathore N Vijayashankar

Abstract A method has been developed, based on the periodate oxidation of the manganese in the coordination product of zinc ion and manganese ethylene bisdithiocarbamate to permanganic acid, to determine DithaneM-45. The relationship between the intensity of color and the concentration of manganese obeys Beer’s law up to 100 μg at 540 nm. The method can be easily applied to the determination of the active ingredient in formulations and also for residue analysis. The method is sensitive to 0.6 μg and can be used to determine 6 /xg fungicide at the 20 g sample level.


2007 ◽  
Vol 90 (1) ◽  
pp. 128-141 ◽  
Author(s):  
Abd El-Maaboud I Mohamed ◽  
Osama H Abdelmageed ◽  
Ibrahim H Refaat

Abstract Simple chemometrics-assisted spectrophotometric methods are described for determination of 2 antibacterial binary mixtures. The mixtures are composed of norfloxacin in combination with tinidazole and erythromycin (as ethylsuccinate ester or stearate salt) in combination with trimethoprim. The normal UV absorption spectra of each pair of drugs in the studied mixtures, in the range of 200-400 nm, showed a considerable degree of spectral overlapping: 77.5% for the norfloxacintinidazole mixture and 84.3% for the erythromycintrimethoprim mixture. Resolution of the norfloxacintinidazole mixture and trimethoprim in the presence of erythromycin was accomplished successfully by using zero-crossing first derivative (1D), classical least-squares (CLS) regression analysis, and principal component regression (PCR) analysis methods. In addition, an alternative simple and accurate colorimetric method was developed for the determination of erythromycin in the presence of trimethoprim using 2,4-dinitrophenylhydrazine. All variables affecting the development of the colored chromogen were studied and optimized, and the product was measured at 526-529 and 538-542 nm for erythromycin stearate and erythromycin ethylsuccinate, respectively. For zero-crossing, first derivative technique Beer’s law was obeyed in the general concentration range of 250 μg/mL for norfloxacin, tinidazole, and trimethoprim with good correlation coefficients (0.9994-0.9996). Overall limits of detection (LOD) and quantification (LOQ) ranged from 0.59 to 2.81 and 1.96 to 9.33 μg/mL, respectively. The obtained results from CLS and PCR were compared with those obtained from a 1D spectrophotometric method. With the exception of erythromycin, overall recoveries in the average range of 97.33-103.0% were obtained with a considerable degree of accuracy when the suggested methods were applied to analysis of synthetic binary mixtures, some commercial dosage forms such as tablets and oral suspension without interference from the commonly encountered excipients and additives. For the colorimetric method, Beer's law was obeyed in the general concentration range of 7.21-28.84 μg/mL erythromycin with good correlation coefficients (0.9980-0.9996). Overall LOD and LOQ ranged from 0.73 to 1.65 and 2.43-5.49 μg/mL, respectively. Erythromycin derivatives were determined in the commercial dosage form, without interference from trimethoprim-encountered excipients and additives. The obtained results, with both chemometric and colorimetric methods, have been compared with those obtained from reported methods, and proper F- and t-values were observed, indicating no significant difference between the results of the suggested methods and reported method(s). The good percentage recoveries and proper statistical data obtained proved the efficiency of the proposed procedures for the determination of the studied drugs in their binary mixtures as well as in the commercial dosage forms with quite satisfactory precision.


1997 ◽  
Vol 80 (6) ◽  
pp. 1368-1373 ◽  
Author(s):  
Kailasam Srtvidya ◽  
Natesan Balasubramanian

Abstract An easy and sensitive spectrophotometric assay of pyridoxine is described. The procedure is based on formation of an azo dye by the reaction of pyridoxine with diazotized 2,4-dinitroaniline followed by the reaction of the dye with Hg2+ ions to form a stable complex with maximum absorbance at 545 nm. The system obeys Beer’s law for 4–75 μg pyridoxine hydrochloride in an overall aqueous volume of 25 mL (correlation coefficient, 0.9998). On extraction into 5 mL butan-1-ol, the system obeys Beer’s law in the range 0.8–15 μg pyridoxine hydrochloride at 545 nm. The color is stable for 60 min in both aqueous and organic phases (molar absorptivity, 3.7 × 104 L/mol · cm; coefficient of variation, 3.1%, n = 10). The pyridoxine contents of pharmaceutical preparations, a processed foodstuff, and 2 rice samples were determined by using the proposed method. Assay reliability was established by recovery studies and parallel determination using a reported method.


1975 ◽  
Vol 21 (6) ◽  
pp. 768-770 ◽  
Author(s):  
Jose Mendez ◽  
Barry Franklin ◽  
Harry Gahagan

Abstract We describe a modified method for determining serum triglycerides (triacylglycerols), which is based on the heptane extraction procedure of Gottfried and Rosenberg [Clin. Chem. 19, 1077 (1973)] with the stable saponification, oxidation, and color development reagents of Neri and Frings [Clin. Chem. 19, 1201 (1973)]. This modified method eliminates one heating step, reduces saponification time to 5 min, absorbances are read at room temperature, and the calibration curve is linear to 3.0 g/liter. A sample comparison between the proposed method and the automated Block and Jarrett [Am. J. Med. Technol. 35, 1 (1969)] procedure showed no significant difference (r = 0.98). The coefficient of variation (47 duplicate samples) for the modified method was 6.3%. Further validation was obtained from analysis of quality-control samples; the proposed method gave equivalent values.


1938 ◽  
Vol 16b (3) ◽  
pp. 97-108 ◽  
Author(s):  
W. J. Dyer ◽  
C. L. Wrenshall

A sensitive and accurate method for the determination of phosphate, involving the application of the Evelyn photoelectric colorimeter to the ceruleomolybdate reaction, is described. This technique makes it possible to differentiate phosphate phosphorus from other forms of phosphorus.Conditions affecting the rate and extent of color development have been studied. The results show that the maximum color intensity is developed m about five minutes after addition of the reducing agent. The determination may be made in the presence of extraneous color, and soil organic matter does not interfere with the reaction. Under the conditions specified, using light filters, Beer's law applies to the reaction in soil extracts as well as in pure solutions in the range 0.02 to 0.40 parts per million of phosphorus.


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