This study aimed to analyze a high-performance liquid chromatography (HPLC) separation using a pentafluorophenyl column of parent drug hydroxychloroquine (HCQ) and its active metabolite, desethylhydroxchloroquine (DHCQ) applying to determine bioequivalence of two different formulations administered to patients. A rapid, simple, sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for bioanalysis of HCQ and its metabolite DHCQ in human whole blood using deuterium derivative hydroxychloroquine-D4 as an internal standard (IS). A triple-quadrupole mass spectrometer was operated using electrospray ionization in multiple reaction monitoring (MRM) mode. Sample preparation involves a two-step precipitation of protein techniques. The removed protein blood samples were chromatographed on a pentafluorophenyl (PFP) column (50 mm × 4.6 mm, 2.6 μm) with a mobile phase (ammonium formate solution containing dilute formic acid) in an isocratic mode at a flow rate of 0.45 mL/min. The standard curves were found to be linear in the range of 2 – 500 ng/mL for HCQ; 2 – 2,000 ng/mL for DHCQ in spite of lacking a highly sensitive MS spectrometry system. Results of intra- and inter-day precision and accuracy were within acceptable limits. A run time of 2.2 min for HCQ and 2.03 min for DHCQ in blood sample facilitated the analysis of more than 300 human whole blood samples per day. Taken together, we concluded that the assay developed herein represents a highly qualified technology for the quantification of HCQ in human whole blood for a parallel design bioequivalence study in a healthy male.

The objectives of this study was to evaluate the degradability and digestibility of crude protein (CP), rumen undegradable protein (RUP), and individual amino acids (AA) on six by-product feedstuffs (BPF) (rice bran, RB; wheat bran, WB; corn gluten feed, CGF; tofu residue, TR; spent mushroom substrate from Pleurotus ostreatus, SMSP; brewers grain, BG) as ruminants feed. Three Hanwoo steers (40 months old, 520 ± 20.20 kg of body weight) fitted with a permanent rumen cannula and T-shaped duodenal cannula were used to examine of the BPF using in situ nylon bag and mobile bag technique. The bran CGF (19.2%) and food-processing residue BG (19.7%) had the highest CP contents than other feeds. The RUP value of bran RB (39.7%) and food-processing residues SMSP (81.1%) were higher than other feeds. The intestinal digestion of CP was higher in bran RB (44.2%) and food-processing residues BG (40.5%) than other feeds. In addition, intestinal digestion of Met was higher in bran RB (55.7%) and food-processing residues BG (44.0%) than other feeds. Overall, these results suggest that RB and BG might be useful as main raw ingredients in feed for ruminants. Our results can be used as baseline data for ruminant ration formulation.

In order to establish a year round indoor-rearing system for Copper butterflies; large copper butterfly, Lycaena dispar, the effect of temperature on larval development and diapause was investigated. Temperature has been suggested as an important factor regulating the developmental rate, length of life, and survival rate from insect. As temperature increased, the developmental period was gradually shorten. The developmental periods of large copper larvae had a range of 11.0 days to 28.5 days at 30℃ and 17.5℃ respectively. The highest emergence rate was 94.2% recorded at 20℃. And the low emergence rate was 72.7% under 17.5℃. We investgated the sensitive stages to diapause induction in the larger copper effect of temperature and photoperiod. The experiment involves transfer of individuals from diapause averting (LD 16:8h, 25℃) to diapause inducing condition (LD 8:16h, 20℃) at various stages. Diapause was induced in 95.2% insect transferred at hatching larvae, in 15.6% of insects transferred at 2nd stadium molt, in 0% of insects transferred at after 3rd stadium molt. Percentage diapause induction increase with the length of short days and low temperature experienced. The main stage sensitive to photoperiod and temperature induction of diapause determination is the early first larval instar. The diapause induction began 14 days and ended 20 days after hatching larvae (LD 8:16h, 20℃).