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High-efficient liquid phase chromatographic analysing method for regulating pyrurine and its impurity

A high-performance liquid chromatography, reversed-phase high-performance liquid phase technology, applied in the field of high-performance liquid chromatography analysis, to achieve the effect of a simple and reliable method

Inactive Publication Date: 2005-01-05
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the method for simultaneously measuring the content of coexisting impurities in products has not been reported yet.

Method used

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  • High-efficient liquid phase chromatographic analysing method for regulating pyrurine and its impurity
  • High-efficient liquid phase chromatographic analysing method for regulating pyrurine and its impurity
  • High-efficient liquid phase chromatographic analysing method for regulating pyrurine and its impurity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1. Selection of chromatographic conditions

[0029] 1.1 Mobile phase

[0030] We compared the composition of different mobile phases and tested the methanol ratio from high to low. Under the condition that the methanol-water ratio is 60:40 (volume ratio), the components can achieve baseline separation, see figure 1 .

[0031] 1.2 Detection wavelength

[0032] Using a diode array detector for continuous scanning at 200-400nm, it can be seen that ACP has a maximum absorption at 244nm, while DPU and CPPU have strong absorption at 254nm. Taking into account the sensitivity of the two determinations, the detection wavelength is selected as 254nm.

[0033] 1.3 Sample concentration

[0034] In order to balance the determination of the main component content and the impurity content, the concentration of the sample must be large enough to make the impurity peak when determining the impurity content; when determining the main component content, the concentration of the solu...

Embodiment 2

[0035] Example 2. Standard curve

[0036] Preparation of standard solution: accurately weigh 100.00mg CPPU reference substance, place it in a 100mL volumetric flask, dissolve and dilute to the mark with mobile phase, shake well to obtain a standard stock solution with a concentration of 1.0mg / mL, and accurately pipette appropriate amounts respectively The stock solution is prepared into a series of standard solutions with mobile phase.

[0037] Accurately weigh 50.00mg ACP reference substance and 50.00mg DPU reference substance, respectively place them in a 100mL volumetric flask, add mobile phase to dissolve and dilute to the mark, shake well to obtain a stock solution with a concentration of 0.50mg / mL. Precisely pipette 10.00ml ACP and 5.00ml DPU stock solution into the same 100mL volumetric flask, add mobile phase to dissolve and dilute to the mark, shake up, the concentration of ACP and DPU are 0.05mg / mL, 0.025mg / mL, respectively Mix the standard stock solution. A series of st...

Embodiment 3

[0044] Example 3. Sample determination

[0045] 3.1 Determination of main content

[0046] Preparation of sample solution: accurately weigh about 100 mg of CPPU sample, place it in a 100 mL volumetric flask, add 10 mL of methanol to dissolve, then add mobile phase to the mark, and shake well for use. Pipette 2.0 mL of the above solution into a 100 mL volumetric flask, add mobile phase to the mark, and shake well for use.

[0047] Select any 5 batches of samples, prepare the sample solution according to the above method, and perform HPLC analysis under the best test conditions. The sample size is 10μL (see Figure 5 ).

[0048] 3.2 Determination of impurities

[0049] Preparation of sample solution: accurately weigh about 100 mg of CPPU sample, place it in a 100-mL volumetric flask, add 10 mL of methanol to dissolve, then add mobile phase to the mark, and shake well for use.

[0050] Select any 5 batches of samples, prepare the sample solution according to the above method, and per...

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Abstract

The invention is a high efficiency liquid phase chromatogram analysis method for forchlorfenuron and its impurities, the high efficiency liquid phase chromatogram condition is: chromatogram pile: C18, 5 micrometer, 150X4.6mm (I.D.): flow phase composition volume proportion: carbinol: water=60:40: flow speed: 1.0mL / min; pile temperature: 30deg.C; the detecting wavelength: UV 254nm; the sample taking quantity: 5-10 micro litre. The method in the invention also can measure the content of CPPU and the impurities ACP and DPU in the CPPU sample. It provides a simple and reliable method for the quality control analysis in the producing process.

Description

Technical field [0001] The invention relates to a high-performance liquid chromatography analysis method, in particular to the use of high-performance liquid chromatography to analyze the content of forchlorfenuron and its impurities. Background technique [0002] N-(2-chloro-4-pyridyl)-N'-phenylurea (abbreviation: forchlorfenuron, N-(2-chloro-4-pyridyl)-N'-phenylurea, CPPU) is a cytokinin Such plant growth regulators can induce the accumulation of anthocyanins and carbohydrates in certain plants such as radishes, tomatoes, apples, berries and grapes. Its physiological activity is higher than that of general purine cytokinin, and it is easy to synthesize and low in cost. It is considered as a cheap substitute for zeatin. The current research on CPPU mainly focuses on its mechanism and application [see: Hou Yong, Ma Guorui, Xia Zhongmei, Zeng Xianbin, Journal of Plant Nutrition and Fertilizer, 1999, 5(2), 106-114]. Regarding the determination method of CPPU, gas chromatography is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/88
Inventor 练鸿振李丹妮朱卫
Owner NANJING UNIV
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