Method for detecting radix astragali

Method for detecting radix astragali

  • CN 104,198,600 A
  • Filed: 07/31/2014
  • Published: 12/10/2014
  • Est. Priority Date: 07/31/2014
  • Status: Active Application
First Claim
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1. a detection method for Milkvetch Root, is characterized in that, the method comprises the determination step of following active constituent content and the determination step of persticide residue, wherein,A:

  • the mensuration of Astragaloside content comprises the steps;

    (1) it is appropriate that precision takes Astragaloside IV reference substance, adds methyl alcohol and make the solution of every 1ml containing 0.5mg, product solution in contrast;

    (2) accurately weighed Milkvetch Root powder 4g to be measured, put in apparatus,Soxhlet'"'"'s, add methyl alcohol 40ml, cold soaking, add again methyl alcohol appropriate, add hot reflux 4 hours, extract reclaims solvent and is concentrated into dry, the residue 10ml that adds water, low-grade fever makes to dissolve, with water saturated normal butyl alcohol jolting extraction 4 times, each 40ml, merge normal butyl alcohol liquid, fully wash 2 times with ammonia solution, each 40ml, discard ammoniacal liquor, normal butyl alcohol liquid evaporate to dryness, residue add water 5ml make dissolve, let cool, adsorb by D101 type large pore resin absorption column, with water 50ml wash-out, discard water liquid, use again 40% ethanol 30ml wash-out, discard eluent, continue with 70% ethanol 80ml wash-out, collect eluent, evaporate to dryness, dissolve and be transferred in 5ml measuring bottle with methyl alcohol, add methyl alcohol to scale, shake up, as need testing solution,(3) according to high performance liquid chromatography test, taking octadecylsilane chemically bonded silica as filling agent;

    Acetonitrile-water taking volume ratio as 32;

    68 is mobile phase;

    Evaporative light-scattering detector detects;

    Accurate reference substance solution 10 μ

    l, the 20 μ

    l of drawing respectively, need testing solution 20 μ

    l, injection liquid chromatography, measures, and calculates with external standard two-point method logarithmic equation;

    Wherein, number of theoretical plate calculates and should be not less than 4000 with Astragaloside IV peak;

    The mensuration of B calycosin glucoside content comprises the steps;

    (1) get calycosin glucoside reference substance appropriate, accurately weighed, add methyl alcohol and make the solution of every 1ml containing 50 μ

    g, product solution in contrast;

    (2) accurately weighed Milkvetch Root powder 1g to be measured, precision adds methyl alcohol 50ml, weighed weight, add hot reflux 4 hours, let cool, more weighed weight, supply the weight of less loss with methyl alcohol, shake up, filter, precision measures subsequent filtrate 25ml, reclaims solvent to dry, residue adds methyl alcohol and dissolves and be settled to 5ml, as need testing solution;

    (3) according to high performance liquid chromatography test, taking octadecylsilane chemically bonded silica as filling agent;

    Taking acetonitrile as mobility A, taking 0.2% formic acid solution as Mobile phase B, carry out gradient elution according to following program;

    from 0-20min, mobile phase A;

    the volume ratio of Mobile phase B is by 20%;

    80% →

    40%;

    60%;

    From 20-30min, mobile phase A;

    the volume ratio of Mobile phase B is 40%;

    60%;

    Controlling and detecting wavelength is 260nm;

    Accurate reference substance solution and the each 10 μ

    l of need testing solution of drawing respectively, note people liquid chromatograph, measures;

    Number of theoretical plate calculates and should be not less than 3000 by calycosin glucoside peak;

    C;

    the mensuration of described persticide residue comprises the steps;

    (1) to take triphenyl appropriate for precision, adds acetone and make the solution of every 1ml containing 100 μ

    g triphenyl, as interior mark stock solution;

    Precision measures each agricultural chemicals reference substance stock solution 1ml and described interior mark stock solution 1ml respectively, adds acetone and is settled to 100ml, as mixing reference substance stock solution;

    Precision measures in right amount respectively, adds acetonitrile constant volume and make the solution of the variable concentrations of 20-1000ng/ml, as mixing reference substance solution;

    Separately get ribose acid lactone appropriate, add acetonitrile dissolving and make the solution of every 1ml containing 20mg ribose acid lactone, separately get sorbierite appropriate, be dissolved in water and make the solution of every 1ml containing sorbierite 10mg;

    Precision measures above-mentioned ribose acid lactone respectively, the each 1ml of sorbitol solution mixes, and adds acetonitrile and is settled to 10ml, as analysis protectant;

    (2) precision takes medicinal material fine powder 10g to be measured, and adds sodium chloride 1g to mix, and precision adds acetone 100ml, and ice-bath ultrasonic is processed 30 minutes, centrifugal, and supernatant is moved into and is equipped with in the tool plug conical flask of 1g anhydrous sodium sulfate rapidly, places 30 minutes;

    Precision measures above-mentioned solution 60ml and is evaporated near doing subsequently, and to add volume ratio be thiacyclohexane-ethyl acetate solution sample dissolution of 1;

    1 and be settled to 10ml, after filtration, getting filtrate purifies through GPC gel permeation chromatography, thiacyclohexane-ethyl acetate solution taking volume ratio as 1;

    1 is mobile phase wash-out, collect eluent, move in KD bottle, be evaporated near dry;

    Be that ethyl acetate-acetone mixed solution 5ml of 1;

    1 dissolves to adding volume ratio in above-mentioned sample, and be transferred on graphitic carbon-amino mixing solid phase extraction column, ethyl acetate-acetone mixed solution 15ml wash-out taking volume ratio as 1;

    1, collect eluent, nitrogen blows near dry, add subsequently described interior mark stock solution 5 μ

    l, add acetonitrile and be settled to 1ml, as need testing solution;

    (3) precision measures above-mentioned each concentration mixing reference substance solution and the each 400 μ

    L of need testing solution, and adds respectively described analysis protectant 100 μ

    L to mix, and precision is drawn 1 μ

    L respectively subsequently, carries out gas chromatograph-mass spectrometer (GCMS) mensuration;

    Wherein,Described analytical conditions for gas chromatography is;

    getting specification is the fused-silica capillary column DB17ms of 30m ×

    0.25mm ×

    0.25um, taking high-purity helium as carrier gas, column flow rate 1.3ml/ minute, sample size 1 μ

    l, adopt high pressure Splitless injecting samples, it is 230 DEG C that injector temperature is set, and heating schedule is specially;

    60 DEG C of initial temperatures, rise to 120 DEG C, rise to 200 DEG C, rise to 230 DEG C, rise to 300 DEG C with 30 DEG C/min with 2 DEG C/min with 10 DEG C/min with 30 DEG C/min, and keep 7 minutes;

    The condition of described EI source mass spectroscopy is;

    electron energy 70eV is set, 230 DEG C of ion source temperatures, 250 DEG C of interface temperature.

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