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#21687539   2011/06/20 Save this To Up

Direct determination of ECD in ECD Kit: a solid sample quantitation method for active pharmaceutical ingredient in drug product.

Technetium-99m ethyl cysteinate dimer (Tc-99m-ECD) is an essential imaging agent used in evaluating the regional cerebral blood flow in patients with cerebrovascular diseases. Determination of active pharmaceutical ingredient, that is, L-Cysteine, N, N'-1,2-ethanediylbis-, diethyl ester, dihydrochloride (ECD) in ECD Kit is a relevant requirement for the pharmaceutical quality control in processes of mass fabrication. We here presented a direct solid sample determination method of ECD in ECD Kit without sample dissolution to avoid the rapid degradation of ECD. An elemental analyzer equipped with a nondispersive infrared detector and a calibration curve of coal standard was used for the quantitation of sulfur in ECD Kit. No significant matrix effect was found. The peak area of coal standard against the amount of sulfur was linear over the range of 0.03-0.10 mg, with a correlation coefficient (r) of 0.9993. Method validation parameters were achieved to demonstrate the potential of this method.

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#8742076   1996/10/11 Save this To Up

Quantitation of L-amino acids by substrate recycling between an aminotransferase and a dehydrogenase: application to the determination of L-phenylalanine in human blood.

A spectrophotometric recycling assay for the quantitation of L-phenylalanine (and phenylpyruvate) has previously been reported (Cooper et al., Anal. Biochem. 183, 210-214, 1989). The procedure involves the coupling of bacterial phenylalanine dehydrogenase with rat kidney cytosolic glutamine transaminase K. The latter enzyme possesses high affinity for phenylpyruvate. Recycling results in a > or = 50-fold increase in sensitivity over that of a conventional spectrophotometric "end point" analysis procedure. The spectrophotometric recycling procedure has now been adapted to the measurement of L-phenylalanine in microliter quantities of human blood. This procedure is 10 times more sensitive than provided by a commercial kit for the spectrophotometric measurement of L-phenylalanine in human blood. Moreover, the present results suggest that the recycling procedure adapted for fluorometry will be even more sensitive. By use of suitable dehydrogenases and amino acid aminotransferases it should be possible to quantitate amino acids (in addition to phenylalanine) in small quantities of human blood.

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#7014033   1981/07/09 Save this To Up

Continuous-flow enzyme immunoassay for thyroxine in serum.

We have evaluated a fully automated thyroxine assay involving the use of a homogeneous enzyme assay and a Technicon AutoAnalyzer II continuous flow system. Comparison by regression analysis with a thyroxine radioimmunoassay kit method gave a slope of 0.82 and a y-intercept of 7.81 micrograms/L (SE 0.86 microgram/L, r = 0.95). Within-run precision yielded CVs of 1.0-6.1%, between-day CVs were 2.0-14.4%; within-day precision showed a mean variance of 7.81 micrograms/L. Mean analytical recovery was 96.1%. Bilirubin, hemoglobin, and lipemia interfered with quantitation of results when their concentrations exceeded 50 mg/L, 300 mg/L, and 5 g/L, respectively. The reference interval for euthyroid status was calculated to be 50-110 micrograms/L. Sensitivity was 5.0 micrograms/L with a mean carryover of 1.65%. Current reagent and labor costs for enzyme immunoassay ($0.40) were less than for the manual radioimmunoassay procedure ($0.40) were less than for the manual radioimmunoassay procedure ($1.70). The assay is economical, simple to perform, and amenable to high-throughput thyroid screening in the routine laboratory.

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