Alexys™ Electrochemistry and Liquid Chromatography Products & Parts Catalogue - Alpha MOS - #9

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The analyzers are dedicated and methods are validated to meet the specifications of the EP and/ or USP . Aminoglycosides such as Gentamicin, Tobramycin, Kanamycin, Spectinomycin and Lincomycin in cocktails can be analyzed. The USP method for Azithromycin utilizes two flow cells in series. The first cell is used for screening, and the second for detection of the analytes. For this application, the ALEXYS Azithromycin Analyzer™ utilizes a FlexCell in series with a VT -03 cell for detection. Proven analyzers to guarantee your performance ® Proven analyzers to guarantee your performance ALEXYS™ Analyzers pharmaceutival & Biotech ALEXYS ™ Analyzers with fluidly running applications ALEXYS Aminoglycosides Analyzer™ • Aminoglycosides • EP /USP methods • B ulk and veterinary cocktails ALEXYS Azithromycin Analyzer™ • Ma crolide Antibiotics • USP method • Applicable for Clari-, Roxi-, and Erythromycine Antec Leyden B.V. - Industrieweg 12 - 2382 NV - Zoeterwoude - The Netherlands - info@myantec.com - www.myantec.com Antec (USA) - Suite 110 - 7502 Connelley Drive - Hanover MD 21076 - USA - info.usa@myantec.com - www.myantec.com Page : 2/4 SPECTINOMYCIN IN PHARMACEUTICAL PREPARATIONS Page : 2/4 217_018 #00 Solutions and standards are prepared as described in the EP method [4,5]. Assay validation was done with special attention to EP requirements. EP requirements Fig. 2. Overlay (n=6) of 20 ìL injections of 80 mg/L Spectinomycin-HCl in mobile phase (diluted from 800 mg/L Spectinomycin in water, standing time 68 h).from paper [2] and based on peak area percentages. In the EP monographs for Spectinomycin [4.5] two system suitability requirements are specified: [1] Peak resolution: between impurity E and the principle peak (Spectinomycin), R > 1.5. [2] Repeatability: maximum RSD(%) for the principle peak, n=6 injections, RSD > 3%. In Table 1 the criteria of the EP are compared with the typical results obtained with the ALEXYS Aminoglycosides analyzer. Parameter EP criteria Result RSD of principal peak Resolution, peak ‘A’ 1.5 1.0% 2.5 It is evident from table I that the EP requirements for both peak resolution and repeatability are met by the ALEXYS aminoglycosides analyzer. Antec Leyden B.V. - Industrieweg 12 - 2382 NV - Zoeterwoude - The Netherlands - info@myantec.com - www.myantec.com Antec (USA) - Suite 110 - 7502 Connelley Drive - Hanover MD 21076 - USA - info.usa@myantec.com - www.myantec.com Page : 3/4 SPECTINOMYCIN IN PHARMACEUTICAL PREPARATION217_018 #00 S CONCLUSION 12 The ALEXYS® Aminoglycosides analyzer provides a reliable solution for the routine analysis of Spectinomycin in Pharmaceutical Preparations. It meets the EP system suitability requirement for resolution and repeatability. Antec Leyden B.V. - Industrieweg 12 - 2382 NV - Zoeterwoude - The Netherlands - info@myantec.com - www.myantec.com Antec (USA) - Suite 110 - 7502 Connelley Drive - Hanover MD 21076 - USA - info.usa@myantec.com - www.myantec.com Page : 2/2 ERROR! REFERENCE SOURCE NOT FOUND. Page : 2/2 217_013 #05 CONCLUSION Due to the high working potential and modifier percentage we recommend to use a flow cell with HyREF (p.n. 110.4305). It appears that the pH of the mobile phase is a decisive factor to warrant the solubility of the buffer salts. We have found that applying a higher pH will result in precipitation. ALEXYS Azitromycin Analyzer™ configurations ALEXYS Azitromycin Analyzer PART NUMBERS AND CONFIGURATIONS 190.0035 ALEXYS 110 LC-EC system with DECADE II SDC 115.4150 SenCell GC, salt bridge, 5/60 12 1 W.R. LACOURSE, “PULSED ELECTROCHEMICAL DETECTION IN HIGH PERFORMANCE LIQUID CHROMATOGRAPHY”, JOHN WILEY & SONS, NEW YORK, 1ED,1997. 2 E. ADAMS, W. ROELANTS, R. DE PAEPE, E. ROETS, J. HOOGMARTENS, J. PHARM. BIOMED. ANAL., 18, 689-698 (1998). 9 Antec Leyden B.V. - Industrieweg 12 - 2382 NV - Zoeterwoude - The Netherlands - info@myantec.com - www.myantec.com Antec (USA) - Suite 110 - 7502 Connelley Drive - Hanover MD 21076 - USA - info.usa@myantec.com - www.myantec.com 217_010 #07 Page : 1/5 BIOTECH & PHARMACEUTICAL APPLICATION NOTE • 1][2][3] [ 4][5] using LC-PAD. The ALEXYS® ‘Aminoglycosides analyzer’ is a dedicated LC solution for the analysis of Neomycin and Framycetin, which matches the EP requirements for peak resolution and signal-tonoise ratio of the principal peak (Neomycin B). In this application note typical results obtained with the Aminoglycosides analyzer are reported demonstrating its performance for the analysis of impurities in bulk drugs. The European Pharmacopoeia (EP) has two monographs describing the analysis of Neomycin and Framycetin [ Method Tabel 1 LC-EC Conditions HPLC ALEXYS aminoglycosides analyzer(part no 180.0050A) Flow rate 0.7 mL/min, post-column: 0.5 mL/min Flow cell Flexcell™ with Au WE and HyREF™ Temperature 32 ..C for separation and detection Range 10 ìA/V ADF 0.5 Hz I-cell -800 - -300 nA NEOMYCIN AND FRAMYCETIN SULPHATE IN BULK DRUGS INTRODUCTION Neomycin is an antibiotic complex consisting of a mixture of the aminoglycosides Neomycin A, B and C, obtained from Streptomyces fradiae, were Neomycin B is the main constituent. It is a widely-used broad spectrum water-soluble antibiotic useful primarily in infections involving aerobic bacteria. It is available as skin ointment (e.g., creams, gels, lotions, etc.) and eye drops. Framycetin (also known as Neomycin B sulphate) is an aminoglycoside antibiotic similar to Neomycin and com-monly sold under the brand name Soframycin. Impurities in neomycin and framycetin preparations are analyzed using reversed phase HPLC, with post-column NaOH addition and pulsed amperometric detection (LC-PAD) [1-3]. THE MOST RELIABLE LC-EC APPLICATIONS FOR PHARMACEUTICAL & BIOTECH ANALYSIS EVER FORMULATED Aminoglycosides Amikacin Framycetin Sulphate Gentamicin Sulphate Kanamycin Sulphate Lincomycin Neomycin Spectinomycin Tobramycin Macrolide antibiotics Azithromycin Azaerythromycin Clarithromycin Erythromycin Roxithromycin Bioanalysis of pharmaceutics Artemisinin, Dihydro-artemisinin, Artemether Etoposide 8-OH-DPAT mesna BNP7787 Vincristine Antec Leyden B.V. - Industrieweg 12 - 2382 NV - Zoeterwoude - The Netherlands - info@myantec.com - www.myantec.com Antec (USA) - Suite 110 - 7502 Connelley Drive - Hanover MD 21076 - USA - info.usa@myantec.com - www.myantec.com 217_018 #00 Page : 1/4 BIOTECH & PHARMACEUTICAL APPLICATION NOTE • Aminoglycosides des • EP/UPS methods • Bulk and veterinary cocktails Method with the reported, demonstrating In this application note typical results obtained ALEXYS® Gentamycin analyzer based on a C18 column are its performance for the analy-sis of Gentimicin. Table 1 LC-EC Conditions HPLC ALEXYS Aminoglycosides analyzer (part no.180.0050A) Flow rate 1 mL/min, post-column: 0.5 mL/min Flow cell Flexcell™ with Au WE and HyREF™ Temperature 35 °C for separation and detection Range 2 ìA/V ADF 0.5 Hz I-cell about 2 ìA Solutions and standards are prepared as described in the EP method [4,5]. Assay validation was done with special attention to EP requirements. Spectinomycin SPECTINOMYCIN IN PHARMACEUTICAL PREPARATIONS INTRODUCTION Spectinomycin is an aminoglycoside- like antibiotic produced by Streptomyces spectabilis. In solution, spectinomycin will undergo a ring opening and closing of the hemiketal function, resulting in an equilibrium mixture of four possible anomers. Hydrolysis with acid produces actinamine and in basic solutions actinospectinoic acid (ASA) is formed. Important fermentation impurities are dihydrospectinomycin and dihydroxyspectinomycin [1, 2].Because of the presence of glycoside groups in Spectinomycin and by-products, LC with pulsed amperometric detection (PAD) has been applied for analysis [3]. Conditions are in correspondence with EP requirements [4,5]. The ALEXYS® ‘Aminoglycosides analyzer’ is a dedicated LC solution for the analysis of Spectinomycin, which matches the EP requirements for peak resolution and repeatability of the principal peak. In this application note typical results obtained with the Aminoglycosides analyzer are reported demonstrating its performance for the analysis of impurities in Spectinomycin bulk drugs. THE MOST RELIABLE LC-EC APPLICATIONS FOR PHARMACEUTICAL & BIOTECH ANALYSIS EVER FORMULATED Aminoglycosides Amikacin Framycetin Sulphate Gentamicin Sulphate Kanamycin Sulphate Lincomycin Neomycin Spectinomycin Tobramycin Macrolide antibiotics Azithromycin Azaerythromycin Clarithromycin Erythromycin Roxithromycin Bioanalysis of pharmaceutics Artemisinin, Dihydro-artemisinin, Artemether Etoposide 8-OH-DPAT mesna BNP7787 Vincristine Antec Leyden B.V. - Industrieweg 12 - 2382 NV - Zoeterwoude - The Netherlands - info@myantec.com - www.myantec.com Antec (USA) - Suite 110 - 7502 Connelley Drive - Hanover MD 21076 - USA - info.usa@myantec.com - www.myantec.com 217_013 #05 Page : 1/2 BIOTECH & PHARMACEUTICAL APPLICATION NOTE • Macrolides Antibiotics • USP method • Others like Clari-, Roxy-, and Erithromycine Method Mobile phase conditions were optimised towards the use of a single working electrode, a low background current and a standard reversed phase column. It appeared that the conditions listed below fulfilled these requirements resulting in highly sensitive, selective and reproducible analyses. Stock solutions of Azithromycin (USP) were made up in 100% in methanol to a final concentration of 200 ìM. Standards for a calibration curve were obtained by diluting this solution 200, 2000 times with mobile phase. To analyse the azithromycin amount in a pharmaceutical preparation, Zithromax tablets (Pfizer) with a stated weight and content of 1,000 and 500 mg resp. were weighed and crushed to powder. A powder fraction 300 mg was dissolved in a final volume of 200 ml methanol in an ultrasonic bath at 30°C during 15 min. A 2 ml fraction of the suspension was diluted 5 times with methanol and 500 times further diluted with mobile phase, filtered over a Gelman nylon Acrodisk 0.2 ìm filter and analysed. Table 1 LC-EC Conditions Column Waters Spherisorb ODS2 100 x 4.6 mm, 3 ìm Flow rate 1.0 mL/min Sample 20ìl Mobile phase 200 mL of 50 mmol/L phosphoric acid and 2 mmol/L KCl is set to pH 5.0 (by NaOH). Then 800 mL methanol is added (final concentration of 80 % MeOH). NB: At a higher pH value precipitation of buffer salts occurs. Azithromycin AZITHROMYCIN IN PHARMACEUTICAL DOSAGE FORMS INTRODUCTION Azithromycin belongs to the macrolides and is an acid stable erythromycin derivative used for antibacterial infections. It has quite favourable pharmacokinetic properties and is highly effective against respiratory tract infections, sexually transmitted diseases and nonclassical pathogens like Heliobacter pylori. Azithromycin has been analysed in biological fluids and in pharmaceutical dosage forms by LC using different detection techniques. Among these, electrochemical detection (EC) excels because of its intrinsic sensitivity and precision. In some cases quite unusual LC-EC conditions (pH, modifier, column materials) have been applied. Also, in the official USP method two amperometric working electrodes in a serial arrangement have been prescribed. We have determined the LC and EC conditions allowing the use of standard reversed-phase column materials, relatively low modifier concentrations, a neutral pH and a single working electrode. THE MOST RELIABLE LC-EC APPLICATIONS FOR PHARMACEUTICAL & BIOTECH ANALYSIS EVER FORMULATED Aminoglycosides Amikacin Framycetin Sulphate Gentamicin Sulphate Kanamycin Sulphate Lincomycin Neomycin Spectinomycin Tobramycin Macrolide antibiotics Azithromycin Azaerythromycin Clarithromycin Erythromycin Roxithromycin Bioanalysis of pharmaceutics Artemisinin, Dihydro-artemisinin, Artemether Etoposide 8-OH-DPAT mesna BNP7787 Vincristine