Times, and a total of about 300 ml of BALF was obtainedįrom each subject. Infused through the work channel of a bronchoscope then Polystyrene balls were obtained from Sekisui Chemical Co.,īronchoalveolar Lavage Informed consent was obtained from all patients. All solutions were prepared using water purified withĪ Milli-Q water system (Millipore Corp., Bedford, U.S.A.). Louis, U.S.A.) ģ,3,5,5-tetramethylbenzidine, Nacalai Tesque Inc.
Serum albumin (HSA), fraction V grade, and goat anti-HSAĪntiserum, Sigma Chemical Co. Obtained from the sources indicated as follows: EriochromeĬyanine R (ECR), Merck AG (Darmstadt, Germany) human Reagents The chemicals used in the present study were This technique was applied to determine traceĪmounts of human albumin in BALF samples.
Human albumin using ECR and visible range as excitation We investigated the fluorimetric determination method of ∗ To whom correspondence should be addressed. Ultraviolet radiation may give rise to fluorescence of other components. Visible excitation and emission spectra and low reagent blankĪt pH <4.0. Saito’s group reported that the HSA-ECR complex showed They used 308 nm as the excitation wavelength. Promising.22) Subsequently, Ci and Chen23) reported a fluorimetric method for the determination of HSA using ECR. Trace albumin, and pointed out that several reagents analogous to CAS including eriochrome cyanine R (ECR) are Method using chromazurol S (CAS) for the determination of Saito et al.20,21) reported a highly sensitive fluorimetric Than the bromocresol green method.18,19) However, theīromocresol purple method is not applicable to BALF samples of low albumin concentration. The bromocresol purple method is also used inĬlinical measurement of HSA, and is more selective for HSA Specificity for HSA in the bromocresol green method is not Lung disorders.9,10) Therefore a sensitive and simple methodįor determining trace amounts of albumin in BALF is desirable.ĭye-binding methods are widely used for the determination of proteins,13) and spectrophotometric methods usingīromocresol green are most commonly used in clinical assays of human serum albumin (HSA).14-17) However, the Lavage fluid (BALF) varies from sample to sample, their concentrations are usually normalized relative to those of albumin.1-6,8) Furthermore, the absolute concentration of albumin itself in BALF may allow characterization of interstitial Into the bronchoalveolar space and aspirated by syringe.1)īecause recovery of the components in bronchoalveolar In bronchoalveolar lavage, sterile saline is administered Of these components is of wide diagnostic and investigative Reynolds and Newball over three decades ago.1) The analysis Of the lower respi ratory tract since the first report of The determination of albumin in BALF samples by the ECR method is useful for investigating lung diseases.Įriochrome cyanine R dye-binding fluorimetry albumin bronchoalveolar lavage fluid interstitial lung diseaseīronchoalveolar lavage has been widely used to recoverĬells, proteins, and small molecules from the alveolar space The albumin level of BALF samples obtained from patients with hypersensitivity pneumonitis was increased versus in healthy subjects. Was lower than that of healthy smokers (n9) (42.9'0.7 and 48.3!5.7 m g/ml, respectively). The albumin level in BALF of healthy nonsmokers (n9) The concentrations of albumin inīALF samples were determined by the ECR method. The ECR method is simpler and more rapid than EIA. In the present study, the results obtained by theĮCR method were compared with those of an EIA, and a good linear correlation was observed between the two High enough to determine trace amounts of albumin in BALF. The ECR-albumin adduct showed intense fluorescence and the calibration curve was linear in the range of 2.5-100 m g/ml of human serum albumin (r0.999). Received Octoaccepted April 12, 2007Ī fluorimetric method using eriochrome cyanine R (ECR) was applied to determine trace amounts of albumin in bronchoalveolar lavage fluid (BALF). Kyoto University Sakyo-ku, Kyoto 606–8507, Japan. Tsushima-Naka, Okayama 700–8530, Japan: and c Department of Respiratory Medicine, Graduate School of Medicine, Pharmaceutical Sciences, Graduate School of Medicine and Dentistry and Pharmaceutical Sciences, Okayama University Osaka University of Pharmaceutical Sciences 4–20–1 Nasahara, Takatsuki 569–1094, Japan: b Department of Takaji SATO,a Yoshihiro SAITO,a Masahiko CHIKUMA,*,a Yutaka SAITO,b and Sonoko NAGAIc 30(7) 1187-1190 (2007)įluorimetric Determination of Trace Amounts of Albumin inīronchoalveolar Lavage Fluid with Eriochrome Cyanine R
0 kommentar(er)
