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A highly sensitive flow injection fluorometry for the determination of albumin was developed and applied to the determination of albumin in human bronchoalveolar lavage fluids (BALF). This method is based on binding of chromazurol S (CAS) to albumin. The calibration curve was linear in the range of 5-200 microg/ml of albumin. A highly linear correlation (r=0.986) was observed between the albumin level in BALF samples (n=25) determined by the proposed method and by a conventional fluorometric method using CAS (CAS manual method). The IgG interference was lower in the CAS flow injection method than in the CAS manual method. The albumin level in BALF collected from healthy volunteers (n=10) was 58.5+/-13.1 microg/ml. The albumin levels in BALF samples obtained from patients with sarcoidosis and idiopathic pulmonary fibrosis were increased. This finding shows that the determination of albumin levels in BALF samples is useful for investigating lung diseases and that CAS flow injection method is promising in the determination of trace albumin in BALF samples, because it is sensitive and precise.

Background: Urinary excretion of desmosine has been reported to be increased in patients with pulmonary fibrosis; however, several investigators have pointed out that measuring urinary desmosine is not a very useful indicator of lung wall destruction. We developed a sensitive time resolved fluoroimmunoassay (TR-FIA) to identify trace amounts of desmosine in bronchoalveolar lavage fluid (BALF), and applied this method to analyse BALF samples from healthy subjects and patients with interstitial lung diseases., Methods: In the proposed TR-FIA, a polystyrene strip was coated with desmosine-conjugated gelatin. The strip was then incubated with rabbit anti-desmosine antibody and the test solution. The desmosine bound to the solid phase and free desmosine in the sample or standard solution were allowed to compete to bind to the anti-desmosine. The solid-phase antibody was detected by Eu-complex conjugated anti-rabbit IgG., Results: The detectable limit of desmosine was 50 fmol/ml in the TR-FIA developed in this study. TR-FIA showed low cross-reactivity against amino acids. BALF desmosine levels were significantly higher in patients with idiopathic fibrosis and sarcoidosis compared with healthy subjects., Conclusions: Desmosine levels in BALF may be useful to investigate lung disease.

A fluorimetric method using eriochrome cyanine R (ECR) was applied to determine trace amounts of albumin in bronchoalveolar lavage fluid (BALF). The ECR-albumin adduct showed intense fluorescence and the calibration curve was linear in the range of 2.5-100 microg/ml of human serum albumin (r=0.999). The sensitivity was high enough to determine trace amounts of albumin in BALF. In the present study, the results obtained by the ECR method were compared with those of an EIA, and a good linear correlation was observed between the two methods (r=0.946). The ECR method is simpler and more rapid than EIA. The concentrations of albumin in BALF samples were determined by the ECR method. The albumin level in BALF of healthy nonsmokers (n=9) was lower than that of healthy smokers (n=9) (42.9+/-20.7 and 48.3+/-15.7 microg/ml, respectively). On the other hand, the albumin level of BALF samples obtained from patients with hypersensitivity pneumonitis was increased versus in healthy subjects. The determination of albumin in BALF samples by the ECR method is useful for investigating lung diseases.