Compositional Analysis of Buckwheat Honey

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2. MATERIALS AND METHODS
2.1. Compositional analysis of buckwheat honey
2.1.1. Reagents, chemicals and materials
Standards of cis, trans-abscisic acid, trans, trans - abscisic acid were obtained from Chengdu Biotechnology Company. Benzoic acid, p-coumaric acid, isoferulic acid, gallic acid, methyl syringate, phaseic acid, vanillin, 4-hydroxybenzaldehyde
4-hydroxybenzoic acid were obtain sigma company.
Methanol (analytical grade and HPLC grade) was obtained from Thermo Fisher Scientific Inc. (Fair Lawn, NJ, USA). Formic acid (analytical grade) and acetic acid (HPLC grade) were purchased from J.T. Baker Inc. (Phillipsburg, NJ, USA). Ultrapure water, was purified using a Milli-Q-Integral System (Millipore, Billerica, MA, USA). The Strata-X-A (60 …show more content…

Firstly, The Strata-X-A SPE cartridges prepared using 3 mL of methanol and equilibrated followed with 3mL of ultrapure water. Next, buckwheat honey samples of 30 g were thoroughly mixed with 120 mL of distilled water, and subsequently the solution was adjusted to pH 7.0 with with 5% ammonium (v/v).The buckwheat honey samples were centrifuged with 8000 × g for 10 min to dislodge the solid particles. The supernatants were was passed through the previously conditioned Strata-X-A SPE cartridges. After the samples was loaded, the columns were washed 4 mL of distilled water for Strata-X-A SPE cartridges to remove compounds of buckwheat honey that were not absorbed on the sorbents . Afterwards, the phenolic compounds were eluted with 5 mL of formic acid:methanol (1:9, v/v). The eluate was evaporated with 99.999% nitrogen and then the residue was redissolved in HPLC-grade methanol with 2% acetic acid with 98% methanol (2 ml). The resulting methanol extracts were filtered through a 0.22-μm filter (Millipore, Carrigtowhill, Cork, Ireland) and stored at 4 °C for further analysis by …show more content…

10 % fecal suspension were centrifuged with 300 × g at 4 ℃ for 5 minutes, the supernatants were regard as the original human intestinal bacteria. The composition of culture medium per liter was as follows: 2.0 g yeast extract, 0.1 g NaCl , 0.04 g K2HPO4 , 0.04 g KH2PO4, 0.01 g MgSO4, 0.01 gCaCl2,2.0 g NaHCO3, 0.02 g chlorhematin, 0.5 g cysteine, 0.5 g cholate , 1.0 mg resazurin, 2.0 mL Tween 80 and 10μL vitamin

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