Adenosine-triphosphate purity detection method
A technology for detection of adenosine triphosphate and purity, which is applied in the field of substance detection, can solve the problems of inaccurate ATP detection method, difficulty in ATP detection kit and instrument quality control, and achieve the effect of accurate and reliable detection method
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Embodiment 1
[0046] Embodiment 1 is qualitative to ATP standard substance with nuclear magnetic resonance method
[0047] ATP (adenosine triphosphate) usually exists in the form of disodium salt and contains 3 parts of water of crystallization. The structural formula is shown in the following formula:
[0048]
[0049] The ATP standard substance is qualitatively verified by nuclear magnetic resonance spectroscopy and organic element analysis.
[0050] (1) Instruments and reagents
[0051] Bruker AVANCE III 500M NMR spectrometer.
[0052] Dimethyl sulfoxide, deuterated water.
[0053] (2) Test results
[0054] Dissolve 10mgATP standard substance in 0.3mL D 2 In O, 1024 samples were taken to measure the C13 spectrum.
[0055] Dissolve 5 mg of ATP standard substance in 0.6 mL of dimethyl sulfoxide, and sample 8 times to measure the H1 spectrum.
[0056] According to the nuclear magnetic resonance results, it can be seen that the number and structure of carbon and hydrogen atoms in t...
Embodiment 2
[0057] Embodiment 2 is qualitative with elemental analyzer
[0058] The molecular formula of adenosine triphosphate disodium salt combined with tricrystalline water is: C 10 h 14 o 13 N 5 P 3 Na 2 ·3H 2 O. In order to judge whether the ATP reference material contains other organic substances except a small amount of degradation products, we use two organic element analyzers to detect the content of C, H, and N elements in the ATP reference material. The test results are shown in Table 1:
[0059] Table 1 Detecting results of C, H and N content in ATP standard substance by two elemental analyzers
[0060]
[0061]
[0062] According to the test results of the organic element analyzer, the C, H, and O contents in the sample are very close to the theoretical value of ATP, which proves that the ATP in the ATP sample is in the form of adenosine triphosphate disodium salt with 3 parts of crystal water. According to the content of carbon, hydrogen and nitrogen, no other...
Embodiment 3
[0063] Embodiment 3 Moisture determination
[0064] The ATP standard substance was placed in a desiccator, and after 24 hours of vacuum drying, it was determined by the Karl Fischer method.
[0065] According to "Chinese Pharmacopoeia", the moisture content in the ATP standard substance was determined by the Karl Fischer method. First pre-titrate with anhydrous methanol to the end point, then accurately weigh 3 parts of pure water, titrate to the end point respectively, calculate the average F value of the Karl Fischer reagent (each milliliter of Fischer's solution is equivalent to the weight of water, mg), and save into the titrator.
[0066] Take 6 parts of ATP standard substance samples, each about 0.15g, accurately weighed, dissolve with ethylene glycol-anhydrous methanol (60:40, volume ratio), and titrate to the end point with the calibrated Karl Fischer reagent to obtain The moisture content of the samples was calculated and the average value was calculated. The test ...
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