Development of a novel method based on liquid chromatography–evaporative light scattering detection for the direct determination of streptomycin and Dihydrostreptomycin (cas 128-46-1) in raw materials, pharmaceutical formulations, culture media and plasma

Add time:09/26/2019    Source:infona.pl

A novel method for the non-derivatization liquid chromatographic determination of streptomycin (STR) and Dihydrostreptomycin (cas 128-46-1) (DHSTR) was developed and validated based on evaporative light scattering detection (ELSD). Utilizing a ThermoHypersil BetaBasic C 18 analytical column, evaporation temperature of 50°C and pressure of nebulizing gas (nitrogen) of 3.5bar, the optimized mobile phase was 1.25mLL −1 TFA aqueous solution, in an isocratic mode at a rate of 1.0mLmin −1 . STR was eluted at 5.6min and DHSTR at 7.8min with a resolution of 4.4. Linear calibration curves were obtained from 2 to 120μgmL −1 (r>0.9990) for STR and 2–75μgmL −1 (r>0.9994) for DHSTR, with a LOD equal to 0.7 and 0.5μgmL −1 , respectively. The developed method was applied for the assay of STR and DHSTR (sulfate) in pharmaceutical raw materials and formulations, while the simultaneous direct determination of sulfate was feasible (t R =2.5min, LOD=1.4μgmL −1 , double logarithmic calibration curve in the range of 4–50μgmL −1 , r>0.9998). Modified isocratic mobile phase (H 2 O–ACN, 90:10, v/v, containing 1.25mLL −1 TFA), was used for the determination of streptomycin B impurity in STR sulfate raw material and a gradient mobile phase (H 2 O–ACN containing TFA) was used for the determination of DHSTR in the presence of penicillinG procaine. The developed method was also applied for the assay of commercial formulations (STR powder and DHSTR injection solution and suspension) (%recovery 98–102, %RSD<1.3, n=3×3), for the determination of STR in bacteria culture medium (%recovery 99.6, %RSD=0.8, n=3×3), and for the determination of DHSTR in human plasma (2.0–23.0μgmL −1 ) after solid phase extraction using carboxylate cartridges (%recovery 98.4–101.8, %RSD=3.2, n=3×3).

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