Method for detecting 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 limited application range and narrow linear range, and achieve the effects of wide linear range, simple operation and high accuracy

Active Publication Date: 2017-01-04
益诺思生物技术南通有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 overcome defects such as narrow linear range and limited scope of application of gentamicin C1a in the prior art when detecting concentration

Method used

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  • Method for detecting gentamicin C1a
  • Method for detecting gentamicin C1a
  • Method for detecting gentamicin C1a

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0056] Reference example 2 (precision test)

[0057] Dilute the standard solution with a concentration of 18 mg / ml in Reference Example 1 into a standard solution 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 derivatization and detection were performed with reference to the derivatization reaction steps and HPLC detection method in Example 1. The injection was repeated 5 times, and the RSD was calculated. The results are shown in Table 1.

[0058] Table 1

[0059]

[0060] It can be seen from Table 1 that the derivatization and detection are carried out according to the preparation method ① of the derivatization reagent in Reference Example 1, the derivatization reaction steps, and the HPLC detection method, and the precision is relatively high.

[0061] Reference example 3 (standard product curve drawing)

[0062] Precisely draw the gen...

Embodiment 1

[0066] Embodiment 1 (precision test)

[0067] Dilute the standard solution with a concentration of 18 mg / ml in Reference Example 1 into a standard solution 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 derivatization and detection were performed with reference to the derivatization reaction steps and HPLC detection method in Example 1. The injection was repeated 5 times, and the RSD was calculated, and the results are shown in Table 2.

[0068] Table 2

[0069]

[0070] It can be seen from Table 2 that the derivatization and detection are carried out according to the preparation method ② of the derivatization reagent in Reference Example 1, the derivatization reaction steps, and the HPLC detection method, and the precision is high.

[0071] Embodiment 2 (standard product curve drawing)

[0072] Precisely draw the gentamicin C1a standard ...

Embodiment 3

[0074] The sample to be tested is derivatized and detected according to the preparation method of the derivatization reagent in Reference Example 1, the derivatization reaction steps, and the HPLC detection method. The peak area is the Y axis, the concentration is the X axis, and linear regression is performed. The results are shown in Figure 4 .

[0075] Wherein, when the concentration value on the X-axis is less than or equal to 14.6mg / mL, its concentration value is the concentration value corresponding to its peak area on the standard curve in Example 2; and when the concentration value on the X-axis is greater than 14.6mg / mL, Calculate its concentration value based on its concentration multiple relative to the sample solution with a concentration of 14.6 mg / mL.

[0076] The resulting regression equation: y=0.5732x+187.06, R 2 = 0.9994. The result shows that when the derivatization reagent of the present invention is used to detect samples with lower purity after derivat...

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Abstract

The invention discloses a method for detecting gentamicin C1a, wherein the method comprises the following steps: mixing a to-be-detected sample containing gentamicin C1a with a derivatization reagent evenly and carrying out a derivatization reaction, and then detecting by using a liquid chromatography or liquid chromatography-mass spectrometry detection method. The derivatization reagent contains several components of a solvent, 1,2-phthalic dicarboxaldehyde, mercaptoacetic acid and a buffer solution; the molar ratio of 1,2-phthalic dicarboxaldehyde to mercaptoacetic acid in the derivatization reagent is 1:3.8-1:0.5; the pH of the derivatization reagent is 10-11; the ratio of the molar amount of 1,2-phthalic dicarboxaldehyde in the derivatization reagent to the molar amount of gentamicin C1a detected in the to-be-detected sample in a single detection result is greater than or equal to 3.59. The method for detecting gentamicin C1a has the advantages of simple operation, and relatively high accuracy and wide linear range in detection of the gentamicin C1a concentration in production samples.

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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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
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 益诺思生物技术南通有限公司
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