Method for simultaneously separation determination of content of three aminoglycoside antibiotics

A simultaneous separation and content technology, applied in measuring devices, material separation, analysis of materials, etc., can solve the problems of incomplete derivatization reaction of HPLC-derivatization method, inability to meet the clinical requirements of trace analysis, and insufficient sensitivity of spectrophotometry, etc. Simple method, high sensitivity and good selectivity

Inactive Publication Date: 2016-12-21
CHONGQING MEDICAL UNIVERSITY
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Problems solved by technology

[0009] In view of this, this application is to solve the problems of incomplete derivatization reaction of HPLC-derivatization method, high ex...

Method used

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  • Method for simultaneously separation determination of content of three aminoglycoside antibiotics
  • Method for simultaneously separation determination of content of three aminoglycoside antibiotics
  • Method for simultaneously separation determination of content of three aminoglycoside antibiotics

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Embodiment

[0037] 1. Reagents

[0038] Standards of tobramycin (TOB), etimicin (ETI) and netilmicin (netilmicin, NET) were purchased from China Institute for the Control of Pharmaceutical and Biological Products, and pontamine blue (PSB) was purchased from Chroma company (Germany). Both methanol and acetonitrile were chromatographically pure (Jiangsu Hanbang Company). BR buffer solution consists of 0.04mol·mL -1 h 3 PO 4 、H 3 BO 3 、CH 3 COOH and 0.02mol·mL -1 The NaOH solution is mixed according to a certain ratio to make buffer solutions with different pH values, and the pH values ​​are corrected with a pH meter. Trifluoroacetic acid (TFA) was analytically pure. Ultrapure water was used in the experiment. Tobramycin eye drops, etimicin sulfate injection and netilmicin sulfate injection were purchased from local pharmacies.

[0039] 2. Instrument

[0040]Agilent 1100 system (USA) (G1322A online degassing, G1311A quaternary pump, G1316A column oven, G1313A autosampler, G1321A ...

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Abstract

The invention discloses a method for simultaneously separation determination of the content of three aminoglycoside antibiotics (AGs) (tobramycin, etimicin and netilmicin); the method successfully separates a mixture of the three antibiotics, and also is applied in determination of the content of the AGs in tobramycin eye drops, an etimicin sulfate injection and a netilmicin sulfate injection, and an HPLC-RRS combined technology is used for determination of the AGs. In an experiment, used chromatographic columns comprise Agilent Zorbox SB-C18 column (250 mm*4.6 mm i.d., 5 [mu]m) and a protective column (Security Guard C18, 4*2 mm, Phenomenex). A mobile phase is methanol:water (containing 0.2% TFA)=5:95 (v/v), the HPLC flow rate is 0.50 mL*min<-1>, [lambda]ex=[lambda]em=362 nm, the column temperature is 30 DEG C, and the sample injection volume is 20 [mu]L. The probe is a 2.0*10<-5> mol*L<-1> PSB solution, the pH is 9.5, the single-pump flow rate is 0.2 mL*min<-1>, the reaction pipe length is 250 cm, and the internal diameter is 0.5 mm. A brand-new HPLC detection mode established with RRS as a detection method is established, and is successfully applied to determination of the content of the AGs in different preparations. The application range of the RRS technology is widened, and the brand-new detection mode is provided for HPLC.

Description

technical field [0001] The application belongs to the technical field of pharmaceutical analysis, and in particular relates to a method for simultaneously separating and measuring the contents of three AGs (tobramycin, etilmicin, and netilmicin). Background technique [0002] Aminoglycoside antibiotics (Aminoglycoside antibiotics, AGs) are glycoside antibiotics formed by linking amino sugar and aminocyclic alcohol through an oxygen bridge, including streptomycin, gentamicin, neomycin, tobramycin, and some Newly synthesized aminoglycosides such as etilmicin and netilmicin. Due to their low cost, low toxicity and broad antibacterial spectrum, AGs have become the most widely used antibiotics in clinical practice. At the same time, AGs has certain ototoxicity and nephrotoxicity. In order to ensure the safety of the drug, the dosage range is narrow and the clinical supervision is very strict. [0003] Due to the lack of characteristic ultraviolet absorbing groups of AGs, the ma...

Claims

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02G01N2030/027
Inventor 尚京川王靖雯文瑶贾春燕
Owner CHONGQING MEDICAL UNIVERSITY
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