HPLC (high-performance liquid chromatography) detection method capable of simultaneously detecting contents of six amino acids in donkey-hide glue blood-supplementing preparation
A technology for amino acids and preparations, applied in the field of HPLC determination of six amino acids in donkey glue blood-enriching preparations, which can solve problems such as complex processing methods, and achieve simple sample processing methods, low requirements for operating proficiency, and good reproducibility sexual effect
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Embodiment 1
[0035] Example 1 Preparation of 6 Amino Acid Mixed Standards
[0036]Take appropriate amount of L-glutamic acid reference substance, L-hydroxyproline reference substance, glycine reference substance, L-alanine reference substance, L-arginine reference substance, L-proline reference substance, and accurately weigh , add 0.1mol / L hydrochloric acid solution to make each 1ml contain L-glutamic acid 73.1μg, L-hydroxyproline 86.4μg, glycine 0.1609mg, L-alanine 64.8μg, L-arginine 59.3μg , a mixed solution of 93.9 μg of L-proline to obtain mixed standard substances of 6 kinds of amino acids.
Embodiment 2
[0037] Example 2 Preparation of test solution
[0038] Method 1. Grind donkey glue blood-enriching granules to be tested into powder, take 0.25g, put it in an ampoule bottle, weigh it accurately, add 5ml of 5mol / l hydrochloric acid, put it on an alcohol blowtorch for drawing and sealing, and ultrasonically dissolve it for 30 minutes, 130 ℃ hydrolysis for 1 hour. Let cool, transfer to an evaporating dish, wash with 10ml of water several times, put the washing liquid into an evaporating dish, and evaporate to dryness in a water bath. Add 0.1mol / L hydrochloric acid solution to dissolve the residue, transfer it to a 25ml measuring bottle, add 0.05mol / L hydrochloric acid solution to dilute to the mark, shake well, centrifuge at 3000r / min for 5 minutes and take the supernatant to obtain the solution to be tested.
[0039] Method 2. After grinding the donkey glue blood-enriching granules to be tested into powder, take 0.25g and put it in an ampoule bottle, weigh it accurately, add...
Embodiment 3
[0041] Example 3 derivatization
[0042] Take 2ml each of the above-mentioned standard solution and the solution to be tested, put them in 10ml centrifuge tubes respectively, add 0.1mol / L phenylisothiocyanate (PITC)-acetonitrile solution (take 120μl phenylisothiocyanate in a 10ml volumetric flask 1.0ml, 1mol / L triethylamine-acetonitrile solution (take 1.4ml of triethylamine in a 10ml volumetric flask, dilute with acetonitrile and add to the mark, shake 1.0ml, vortexed for 1min, left at room temperature for 1 hour, added 4ml of n-hexane, shaken, vortexed for 1min, left for 10 minutes, removed the lower layer solution, filtered through a microporous membrane to obtain the derivatized standard solution and derivatized test solution.
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