CN104880444A - Method for determining kanamycin sulfate - Google Patents

Abstract

The invention discloses a method for determining kanamycin sulfate. The method comprises the following steps: adding 0.0, 0.1, 0.5, 0.8, 1.0, 1.2mL of 10.microng/mL kanamycin sulfate solution in 6 10mL colorimetric tubes, adding 0.3-1.5mL of a Britton-Robinson buffer solution with the pH of 6.0 and 0.5-2.0mL of a 2.0*10<-5>mol/L tetra carboxylic-zinc phthalocyanine solution, diluting to the scale by secondary distilled water, and shaking evenly; reacting for 10 minutes at room temperature, and scanning a resonance scattering spectroscopy by a 1cmquartz colorimetric utensil by a mode of lambda ex=lambda em; determining the resonance scattering intensity I332nm of the solution with kanamycin sulfate and the resonance scattering intensity I0 of the reagent blank at the 332nm part, and calculating the difference value delta I332nm=I332nm-I0; and diluting kanamycin sulfate eye drops or kanamycin sulfate injection, determining the resonance scattering intensity by the same method, and calculating the content of kanamycin sulfate in th eye drops or the injection. The method is high in flexibility, is good in selectivity, and is simple, convenient and rapid.

Description

A kind of method for measuring kanamycin sulfate

technical field

The invention relates to a method for determining kanamycin sulfate, in particular to a resonance scattering spectrometry method for determining kanamycin sulfate by using tetracarboxyzinc phthalocyanine.

Background technique

Kanamycin Sulfate (KANA) is a broad-spectrum aminoglycoside antibiotic, mainly effective against severe infections caused by Gram-negative bacteria such as Escherichia Second-line drugs are used to treat tuberculosis and are clinically important anti-infective drugs. However, the abuse of antibiotics will cause serious adverse consequences, and the dosage of antibiotics should be strictly controlled clinically. Therefore, the quantitative determination of kanamycin is of great significance. Commonly used methods for the determination of kanamycin include microbial methods, high performance liquid chromatography, spectrophotometry, optical rotation, colorimetry, electrochemical methods, and resonance scattering spectroscopy. Among them, resonance scattering spectroscopy has the advantages of simple operation, high sensitivity, good selectivity, and rapid analysis. It has been used in pharmaceutical analysis in recent years. The structure of tetracarboxyzinc phthalocyanine (ZnC ₄ Pc) is similar to that of porphyrin, and it can form ionic associations with kanamycin sulfate in weakly acidic solution through electrostatic attraction, which increases the resonance scattering intensity of the system. The method for the determination of kanamycin sulfate by resonance scattering spectrometry has not been reported yet.

Contents of the invention

The object of the present invention is to provide a kind of resonance scattering spectrometry that utilizes tetracarboxyzinc phthalocyanine to measure kanamycin sulfate.

The specific steps are:

Add 0.0, 0.1, 0.5, 0.8, 1.0, 1.2mL 10.0μg/mL kanamycin sulfate solution to six 10mL colorimetric tubes, and then add 0.3-1.5mL pH6.0 Britton-Robinson (Britton-Robinson, BR) buffer solution and 0.5~2.0mL 2.0×10 ^-5 mol/L tetracarboxyzinc phthalocyanine solution, dilute to the mark with twice distilled water, shake well, and react at room temperature After 10 minutes, use a 1cm quartz cuvette to scan the resonant scattering spectrum in the form of λex=λem on the fluorescence photometer; measure the resonant scattering intensity I _332nm of the solution with kanamycin sulfate and the solution without sulfuric acid at 332nm. For the resonance scattering intensity I ₀ of the reagent blank solution of kanamycin, calculate the difference ΔI _332nm =I _332nm -I ₀ , and the difference ΔI _332nm is linear to the concentration C of kanamycin sulfate in the range of 0.1-1.2μg/mL relationship, the linear regression equation is ΔI _332nm =19.43 C-2.201, the correlation coefficient is 0.9922, and the detection limit is 0.029μg/mL; take Kanamycin Sulfate Eye Drops or Kanamycin Sulfate Injection and dilute with water to obtain a concentration of 400μg/mL solution; then dilute the solution to obtain 10μg/mL test solution; take 1-3 mL test solution, measure the resonance scattering intensity with the same method, and calculate the concentration of kanamycin sulfate eye drops or injection The content of kanamycin sulfate.

The determination method of the invention has high sensitivity, good selectivity, simplicity and quickness.

Description of drawings

Fig. 1 is the resonance scattering spectrogram of the embodiment blank and 1 μg/mL kanamycin sulfate of the present invention.

Marks in the figure: a: blank; b: blank + 1 μg/mL KANA.

Detailed ways

Example:

Add 0.0, 0.1, 0.5, 0.8, 1.0, 1.2mL 10.0μg/mL kanamycin sulfate solution to six 10mL colorimetric tubes respectively, and then add 1.0mL pH6.0 Burry to each colorimetric tube Tan-Robison buffer solution and 1.3mL 2.0×10 ^-5 mol/L tetracarboxyzinc phthalocyanine solution, dilute to the mark with twice distilled water, shake well, react at room temperature for 10 minutes, and place in a 1cm quartz cuvette Scan the resonance scattering spectrum in the form of λex=λem on the fluorescence photometer; measure the resonance scattering intensity I _332nm of the solution with kanamycin sulfate and the resonance scattering of the reagent blank solution without kanamycin sulfate at 332nm Intensity I ₀ , calculate the difference ΔI _332nm =I _332nm -I ₀ , the difference ΔI _332nm has a linear relationship with the concentration C of kanamycin sulfate in the range of 0.1-1.2μg/mL, and the linear regression equation is ΔI _332nm =19.43C -2.201, correlation coefficient 0.9922, detection limit 0.029 μg/mL; take another 8 mL of Kanamycin Sulfate Eye Drops into a 100 mL volumetric flask or 1 mL of Kanamycin Sulfate Injection into a 250 mL volumetric flask, and dilute with water to the mark, shake well. Then take 2.5mL of the above solutions respectively into 100mL volumetric flasks, add water to dilute to the mark, shake well, and use it as the solution to be tested. Take 1mL of the above liquid to be tested to measure the resonance scattering intensity in the same way, and carry out the recovery experiment, the results are shown in Table 1.

Determination results of kanamycin sulfate samples (n=5)

sample Measured value (g/L) Labeled amount (g/L) Recovery rate( %) RSD(%) eye drops 4.98 5 108.3 1.1 Injection 243.0 250 104.3 0.25

Claims (1)

1. a method for measuring kanamycin sulfate, is characterized in that concrete steps are: Add 0.0, 0.1, 0.5, 0.8, 1.0, 1.2mL 10.0μg/mL kanamycin sulfate solution to six 10mL colorimetric tubes, and then add 0.3-1.5mL pH6.0 Britton-Robinson buffer solution and 0.5~2.0mL 2.0×10 ^-5 mol/L tetracarboxyzinc phthalocyanine solution, dilute to the mark with twice distilled water, shake well, react at room temperature for 10 minutes , use a 1cm quartz cuvette to scan the resonance scattering spectrum in the form of λex=λem on the fluorescence photometer; measure the resonance scattering intensity I _332nm of the solution with kanamycin sulfate and the solution without kanamycin sulfate at 332nm The resonance scattering intensity I ₀ of the reagent blank solution of the prime element, the calculated difference ΔI _332nm =I _332nm -I ₀ , the difference ΔI _332nm has a linear relationship with the concentration C of kanamycin sulfate in the range of 0.1-1.2μg/mL, linear The regression equation is ΔI _332nm =19.43 C-2.201, the correlation coefficient is 0.9922, and the detection limit is 0.029μg/mL; take another Kanamycin Sulfate Eye Drops or Kanamycin Sulfate Injection and dilute it with water to obtain a concentration of 400μg/mL Then dilute the solution to obtain 10 μg/mL test solution; take 1-3 mL test solution, measure the resonance scattering intensity with the same method, and calculate the concentration of kanamycin sulfate eye drops or kanamycin sulfate injection. The content of mycotoxin.