7150 automated analyzer

A HITACHI 7150 automated analyzer was used for assessment of HNA using the optimized BCP reagents. Two reagents (reagent A and B) were prepared. Reagent A, used to measure total albumin, was prepared according to the method of Muramoto et al.^{(13 (link))} The final concentration of reagent A is mentioned in Table 1. We then searched for the optimal condition for developing the new reagent B that would be the most sensitive for HNA. When we developed the reagent B, we specifically varied the type and concentration of the surfactant in the reagents, pH of the buffer solution, sample volume, and BCP dye concentration to find the combination with maximum sensitivity to HNA, and determined the reagent composition in this order. The final concentration of reagent B is mentioned in Table 2. Measurement for each sample was done in the same way. First, 180 µl of reagent 1 (R1) was added to the serum and left to react at 37°C after stirring for 5 min; this was followed by the addition of 90 µl of reagent 2 (R2). After reacting at 37°C, with stirring for 5 min, the absorbance was measured at 605 nm (main) and 660 nm (sub) (Fig. 1). For comparison, the HNA levels in the same samples were determined using HPLC according to the method described by Sogami et al.^{(3 (link))}

Participants were interviewed to collect sociodemographic data. Height was measured using a stadiometer and weight was measured using an InBody 720 scale (Biospace Corporation, Seoul, Korea). Blood samples were collected after 8 h of overnight fasting, and aliquots of serum were stored at -80℃. Blood chemistry analysis of serum for glucose, total protein, total cholesterol, GOT, and GPT levels was performed using a Hitachi 7150 automated analyzer (Hitachi Ltd, Tokyo Japan) at the Korea Biomedical Laboratory.