A method is developed to determine salbutamol in human plasma and urine using high-performance liquid chromatography (HPLC) with a coulometric electrode array system, based on the electrochemical behavior of salbutamol at graphite electrode. The mobile phase component A is 30 mM sodium dihydroxy phosphate-30 mM triethylamine and is adjusted to pH 6.0 with 20% phosphate acid. The mobile phase component B is methanol. The optimized mobile phase composition was A and B in the proportion of 90:10 (v/v). Paracetamol is selected as the external standard. The human plasma and urine samples are pretreated using solid-phase extraction cartridges (Sep-Pak Silica), and the eluting solution is monitored by the coulometric electrode array system. The electrode potentials are set at 300, 400, 550, and 650 mV, respectively. Calibration curves show good linearity, and the recovery of salbutamol proves to be constant and unaffected by the concentration of the drug. This method, developed using HPLC-electrochemical detection, is reproducible and sensitive enough for the determination of salbutamol in human plasma and urine.
This study investigated the effects of tannic acid (TA)-treated corn on changes in ruminal fermentation characteristics and the composition of the ruminal bacterial community in vitro. Ruminal fluid was obtained from three rumen-fistulated goats fed a 60:40 (forage/concentrate) diet. The batch cultures consisted of 25 ml of strained rumen fluid in 25 ml of an anaerobic buffer containing 0.56 g of ground corn, 0.24 g of soybean meal, 0.10 g of alfalfa, and 0.10 g of oat grass. Ground corn (2 mm) was steeped in an equal quantity (i.e., in a ratio of 1:1, w/v) of water alone (Con), 15 (TA15), 25 (TA25), and 35 g/l (TA35) TA solution for 12 h. After incubation for 24 h, TA-treated corn linearly increased (P <0.05) ruminal pH and the molar proportion of acetate, but linearly reduced (P <0.05) total volatile fatty acids and the molar proportion of butyrate compared with the Con treatment. Illumina MiSeq sequencing was used to investigate the profile changes of the ruminal microbes. A principal coordinates analysis plot based on weighted UniFrac values revealed that the structure of the ruminal bacterial communities in the control group was different from that of the TA-treated corn groups. The results of changes in the rumen bacterial communities showed that TA-treated corn linearly enriched (P <0.05) Rikenellaceae_RC9_gut_group, but linearly reduced (P <0.05) Ruminococcaceae_NK4A214_group, Ruminococcus_2, and unclassified_o__Clostridiales. Functional prediction of ruminal microbiota revealed that the TA-treated corn linearly decreased ruminal microbiota function of utilizing starch through pyruvate metabolism. In conclusion, TA-treated corn can modulate the rumen fermentation characteristics, microbial composition, and metabolic pathways, which may be potentially useful for preventing the occurrence of ruminal acidosis.
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