Detection method of gentamicin C1a

A technology for the detection of gentamicin and its detection method, which is applied in the field of detection of gentamicin C1a, can solve the problems of narrow linear range and limited application range, and achieve the effects of wide linear range, high accuracy and simple operation

Inactive Publication Date: 2017-01-04
SHANGHAI INST OF PHARMA IND +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The technical problem to be solved in the present invention is to provide a detection method for gentamicin C1a in order to ov...

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  • Detection method of gentamicin C1a
  • Detection method of gentamicin C1a
  • Detection method of gentamicin C1a

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0045] Reference example 2 (precision test)

[0046] Dilute the standard solution with a concentration of 18 mg / ml in Reference Example 1 into a standard solution with a concentration of 1.155 mg / ml with distilled water; then prepare the derivatization reagent according to the preparation method of the derivatization reagent in Reference Example 1, and follow the reference The derivatization reaction step and HPLC detection method in Example 1 were used for derivatization and detection. The injection was repeated 5 times, and the RSD was calculated. The results are shown in Table 1.

[0047] Table 1

[0048]

[0049] It can be seen from Table 1 that the derivatization and detection are carried out according to the preparation method of the derivatization reagent, the derivatization reaction steps, and the HPLC detection method in Reference Example 1, and the precision is relatively high.

[0050] Reference example 3 (standard product curve drawing)

[0051] Precisely dra...

Embodiment 1

[0055] Embodiment 1 (measure the concentration of gentamicin C1a after the sample to be tested is diluted)

[0056] Precisely draw the solution of the sample to be tested in Reference Example 1, and dilute it into solutions of different multiples, such as: 1, 1.2, 1.4, 1.6, 1.8, 2, 4, 6, 8, and 10 times. According to the preparation method of the derivatization reagent in Reference Example 1, the derivatization reaction steps, and the HPLC detection method, derivatization and detection are carried out, and the peak area obtained from the detection is found according to the standard curve in Reference Example 3. Corresponding concentration value, the The concentration of the sample to be tested can be obtained by multiplying the concentration value by the dilution factor corresponding to the sample to be tested.

[0057] Take the dilution factor as the X-axis, and the concentration of the sample to be tested as the Y-axis, and draw a curve. The results are shown in image 3 . ...

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Abstract

The invention discloses a detection method of gentamicin C1a. The method comprises the following steps: evenly mixing a gentamicin C1a-containing sample to be tested with a derivatization reagent, and carrying out a derivatization reaction; after that, detecting by a method of liquid chromatography or liquid chromatography-mass spectrometry, wherein the derivatization reagent contains the following components: a solvent, phthalic dicarboxaldehyde, mercaptoacetic acid and a buffer solution; the molar ratio of the phthalic dicarboxaldehyde to the mercaptoacetic acid in the derivatization reagent is (1to 8)-(1to 3.89); the pH value of the derivatization reagent is equal to 10-11; the ratio of the molar quantity of the phthalic dicarboxaldehyde in the derivatization reagent used in a single detection result to the molar quantity of the gentamicin C1a obtained by testing the sample to be tested needs to be more than or equal to 1.6. The detection method of the gentamicin C1a is simple in operation; the detection method is higher in accuracy and wide in linear range when being used for detecting the concentration of the gentamicin C1a in a production sample.

Description

technical field [0001] The invention relates to a detection method of gentamicin C1a. Background technique [0002] The existing HPLC pre-column derivatization detection method for gentamicin C1a is generally the detection method for gentamicin sulfate in the USP35-nf30 edition. For example, accurately measure 10mL of sample into a volumetric flask, add 5mL of isopropanol and 4mL of LOPA derivatization reagent and mix well, then dilute to 25mL with isopropanol, mix well and seal it, place in ice water after 60℃ water bath for 15min to terminate After the reaction, centrifugation at 12000rmp for 5 minutes can be performed for HPLC detection. Wherein, the OPA derivatization reagent formula used: o-phthalaldehyde 1g (0.0074mol), be dissolved in 5ml methanol, add thioglycolic acid 2ml (0.0288mol), finally add 0.4mol / L boric acid (45% sodium hydroxide adjustment pH value 10.4) 95ml and mix well, then adjust the pH value to 10.4 with 45% sodium hydroxide, and store in the refrig...

Claims

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Application Information

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IPC IPC(8): G01N30/88G01N30/06
Inventor 李继安林惠敏陈齐丁碧粉张韬李亚军张羡媛刘一阳张骏梁张建斌
Owner SHANGHAI INST OF PHARMA IND
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