High performance liquid chromatography analysis method for 2,2-2(3-amino-4-hydroxyl radical) phenyl propane and its impurity content
A high-performance liquid chromatography, phenylpropane technology, applied in the direction of analyzing materials, measuring devices, material separation, etc., to achieve the effect of simple and reliable analysis methods
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Embodiment 1
[0036] Embodiment 1: the selection of high performance liquid chromatography condition
[0037] 1.1 Selection of mobile phase
[0038] Secondary quartz distilled water (containing phosphoric acid-disodium hydrogen phosphate buffer solution with pH 7.0) and acetonitrile were mixed in proportion as the mobile phase, and the samples and standard solutions were analyzed. The results show that when the acetonitrile ratio is less than 55%, 2,2-bis(3-nitro-4-hydroxyl) phenylpropane and 2,2-bis(3-amino-4-hydroxyl)phenylpropane cannot achieve complete separation. In order to take into account the separation degree and the measurement time simultaneously, the ratio of acetonitrile-water (phosphoric acid-small diammonium hydrogen phosphate buffer solution containing pH7.0) is selected to be 70:30ml / ml (volume ratio). At this time, each component can realize Baseline separation, and analysis time and reasonableness, see Figure 1-b .
[0039] 1.2 Selection of detection wavelength
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Embodiment 2
[0043] Embodiment 2: the preparation of standard solution and sample solution
[0044] Accurately weigh 25.00mg of 2,2-bis(4-hydroxy-3-nitro)phenylpropane reference substance, place it in a 50ml volumetric flask, dissolve and dilute to the mark with mobile phase, shake well, and obtain a concentration of 0.5mg / ml of 2,2-bis(4-hydroxy-3-nitro)phenylpropane standard stock solution, when used, gradually dilute to the required concentration.
[0045] Accurately weigh 25.00mg of 2,2-bis(3-amino-4-hydroxy)phenylpropane reference substance, place it in a 50ml volumetric flask, dissolve and dilute to the scale with mobile phase, shake well, and obtain a concentration of 0.5mg / ml of 2,2-bis(3-amino-4-hydroxyl) phenylpropane standard stock solution, when used, gradually dilute to the required concentration.
Embodiment 3
[0046] Embodiment 3: the making of standard curve
[0047] Choose the best HPLC conditions, that is, the column is C 18 , 5 μ m, 150 * 4.6mm (I.D); The volume ratio that mobile phase is made up is: acetonitrile: phosphoric acid-disodium hydrogen phosphate buffer solution (pH7.0)=70: 30, ml / ml; Flow velocity is 0.8ml / min ; The column temperature is 28°C; the detection wavelength is 285nm.
[0048] Take 2,2-bis(3-nitro-4-hydroxyl) phenylpropane standard solution 15 μl sample injection, plot the concentration with the peak area ( image 3 ), 2,2-bis(3-nitro-4-hydroxy)phenylpropane solution at a concentration of 2.2×10 -3 In the range of ~1.0mg / ml, the sample concentration has a linear relationship with the peak area, and the linear regression equation is:
[0049] A 2,2-two (3-nitro-4-hydroxyl) phenylpropane=3.6307×10 2 +3.6843×10 4 C, r=0.99807
[0050] Take 2,2-two (3-amino-4-hydroxyl) phenylpropane standard solution 15 μl sample injection, plot the concentration with the...
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