d(GC) - Association of Biotechnology and Pharmacy

More documents

Recommendations

Info

Current Trends in Biotechnology and Pharmacy Vol. 6 (2) 196-203 April 2012, ISSN 0973-8916 (Print), 2230-7303 (Online) Simple, Rapid and Sensitive Method for Determination of Nitrofurantoin in Human Plasma by using Liquid Chromatography / Tandem Mass Spectrometry Rajaram S. Patil 1 *, Chaitanya Krishna, A 2 ., P. V.D.L.S.Ravi Prakash 2 and Sangita R. Patil 2 . 1 Thermofisher Scientific India Pvt. Ltd. Mumbai, India 2 Quest life sciences Private Ltd, Chennai, India * For Correspondence - drrsp1@rediffmail.com Abstract A simple, rapid and sensitive method using an isocratic Liquid chromatography coupled with Tandem mass spectrometry was developed and validated for the assay of nitrofurantoin in the Human Plasma. The Mass transition of nitrofurantoin and losartan (Internal standard) were M/z 237.000/151.800 and M/Z 421.300/ 179.274 in ESI Negative ionization. Linearity was observed between the nitrofurantoin concentration and the peak area ratio from 10.107 to 999.900ng/mL with r 2 value of 0.99. Plasma samples containing nitrofurantoin were extracted with ethyl acetate. The observed recovery of nitrofurantoin was 80.2 %. The intraday and inter-day accuracy ranged from 83.61 to 107.16% and from 93.13to 103.02% respectively, at Low, middle and high level concentrations. The method will be used in the determination of the pharmacokinetic parameters of nitrofurantoin after oral administration of nitrofurantoin formulation in human Plasma. Keywords: Nitrofurantoin, LCMS/MS, Bioanalytical, Validation, Human Plasma, 5nitrofurans Introduction Nitrofurantoin is an antibiotic used for the treatment and prophylaxis for urinary tract 196 infections. Nitrofurantoin is more soluble in urine due to the presence of urea and creatinine. Considerable amount of nitrofurantoin and its metabolites are excreted in urine. This may be the reason; possibly, nitrofurantoin is contraindicated in patients with a creatine clearance of 60mL/min or less, though it is not contraindicated in rest of the population. (6, 21) Therefore, regular monitoring is required in patients receiving nitrofurantoin. Several methods are reported for routine drug monitoring by using different analytical methods such as LC- MS/MS and other conventional methods (1-20). The authors suggest that performing analysis by LC-MS/MS (3) includes derivatisation techniques and is no longer beneficial with the recent advances in the techniques available. There is a need for an established method for analysis which is not tedious and cost consuming, as derivatisation requires more reaction time as well as more amounts of solvents and chemical consumption. More over it is also required to see that the new methods being developed does not require more retention time so that analysis is simple and faster. Therefore, authors proposed a simple, sensitive, accurate and precise method for determination of nitrofurantoin by LC-MS/MS method for measuring nitrofurantoin plasma concentration. Nitrofurantoin in Human Plasma by using Liquid Chromatography / Tandem Mass Spectrometry
Current Trends in Biotechnology and Pharmacy Vol. 6 (2) 196-203 April 2012, ISSN 0973-8916 (Print), 2230-7303 (Online) Stricture of Nitrofurantoin Materials and Methods Materials: Nitrofurantoin and Losartan analytical standards were obtained from Biocon India Pvt Ltd, HPLC grade Acetonitrile and methanol were obtained J.T. Baker, USA, Ammonium acetate (GR Grade Merck), Dimethyl sulphoxide (AR Grade Merck), Orthophosphoric acid (GR Grade Merck),Glacial acetic acid(GR Grade Merck ),Water was de-ionized and triple distilled. Preparation of Nitrofurantoin standard solutions: Standard solutions of Nitrofurantoin Plasma Sample Thaw (30 min) Centrifuge at 4500 rpm for 10 minutes at 5°C Collect 1.8 mL of supernatant Add 50 µL of ISTD Instrumentation and conditions : The Thermo Scientific HPLC (Model: Surveyor) and LC-MS/ MS system interfaced to LC Quan Software in a windows platform The Reverse phase C8 analytical column (Kromosil C 8, Length: 4.6×50 mm, Particle size: 5 µ) was used for the separation of Nitrofurantoin and internal standard, the mobile phase consists a mixture of 197 (1.0 mg/ml) was prepared by dissolving an accurately weighed amount of 10.00 mg of Nitrofurantoin in 5 ml of Acetonitrile and add 50 µL of Dimethyl sulphoxide, sonicated for 5 minutes and the volume was adjusted to 10 ml with Acetonitrile. The standard solution was stored subsequently at 2-8 0C. The appropriate concentrations of standard solution were prepared by diluting the stock solution with 80: 20 Acetonitrile: water. Preparation of Losartan Internal standard solutions: Standard solution of Losartan (1.0 mg/ ml) was prepared by dissolving an accurately weighed amount of 10.00 mg of Losartan in 5 ml of methanol, sonicated for 5 minutes and the volume was adjusted to 10 ml with methanol. The standard solution was stored subsequently at 2- 8 0C. 10.00µg/mL Losartan was prepared by diluting the stock solution with 80: 20 methanol: water. Sample preparation Vortex for 5 min Vortex (1 min) Acidfy with 50µL of 10 % Orthophosphoric acid Vortex (1 min) Add 2.5 mL Ethyl Acetate Evaporate the supernatant at 40 0 C With Nitrogen Reconstitute the residue with 5mM Ammonium Acetate: Acetonitrile (50:50%) (50:50%) and inject 10ul into LC-MS/MS. Rajaram S. Patil et al Acetonitrile and 5mM Ammonium Acetate at a ratio of 60:40 respectively. The mobile phase was degassed by passing through a 0.22 µm membrane filter (Millipore, Bedford, MA, USA) prior to use and operated through a single pump (Isocratic) at a flow rate of 0.5 ml/min. The injector was filled with an injector loop of 10 µl. Tandem mass spectrometric detection and quantification
Page 3 and 4:
Current Trends in Biotechnology and
Page 5 and 6:
Page 7 and 8:
Page 9 and 10:
Page 11 and 12:
Page 13 and 14:
Page 15 and 16:
Page 17 and 18:
Page 19 and 20:
Page 21 and 22:
Page 23 and 24:
Page 25 and 26:
Page 27 and 28:
Page 29 and 30:
Page 31 and 32:
Page 33 and 34: Current Trends in Biotechnology and
Page 83: Current Trends in Biotechnology and
Page 135 and 136:
Page 137 and 138:
Page 139 and 140:
Page 141 and 142:
Page 143 and 144:
Page 145 and 146:
Page 147 and 148:
6th Annual Convention of Associatio
show all

d(GC) - Association of Biotechnology and Pharmacy

Create successful ePaper yourself

Delete template?

Save as template?