Asseptic acid content determination

I, principle
also prototype astrochmic acid (AsA) can reduce iron ions into iron ions, ferrel ions and red ferroids (4,7-dyl-1, 10-figgo, BP) reaction to form a red chelate. Absorption values at 534nM wavelengths are positively related to AsA content, so they can be determined by color ratio. Deoxygenated astrophic acid (DAsA) can be restored to AsA by
(
DTT). The total amount of AsA is determined, from which the prototype AsA is subtracted, i.e. the DASA content.
II, instruments and appliances
centrifuges
;
photonometers
; research;
test tube
.
,
reagents
5% tachloroacetic acid (TCA); 20% TCA; aqueous ethanol solution; 0.4% phosphoric acid-ethanol solution; 0.5% BP-ethanol solution; 0.03% FeCl 3-ethanol solution; 0.6g/L DTT; Na2HPO4-NaOH solution: mixed in 0.2mol/L Na2HPO4 and 1.2mol/L NaOH equivalent; 60mmol/L DTT-ethanol.
fourth, method
1. Production standard curve preparation concentration of 2mg/L, 4mg/L, 6mg/L, 8mg/L, 10mg/L, 12mg/L, 14mg/L AsA series standard liquid. Take each concentration of standard liquid 1.0 ml in the test tube, add 1.0 ml 5% TCA, 1.0 ml ethanol shake, and then add 0.5 ml 0.4% H3PO4-ethanol, 1.0 ml 0.5% BP-ethanol, 0.5 ml 0.03% FeCl3-ethanol, total volume 5.0ml. The solution is placed at 30 degrees C to react 90min, and then the A534 is determined. With AsA concentration as the horizontal coordinate, the standard curve is drawn with A534 as the ordinate, and the line-out equation is sewn.
2. Extract plant leaves 1.0g, add 5% TCA grinding at 1:5 (W/V), 4000r/min centrifugation 10min, and liquid for determination.
3. The determination of
(1) AsA to take 1.0 ml of sample extract in a test tube is determined according to the same method as above, and the AsA content is calculated according to the standard curve.
(2) DAsA assay adds 0.5 ml of 60mmol/L DTT-ethanol solution to 1.0ml sample fluid, uses Na2HPO4-NaOH mixture, adjusts the solution PH to 7 to 8, places it at room temperature 10min, and restores DASA. Then add 0.5ml 20% TCA and adjust the PH to 1 to 2. The total AsA content is calculated by the same method as AsA, from which asA is subtracted, i.e. the DASA content is extracted.
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