OPTIMISATION AND VALIDATION OF HPLC METHOD FOR SIMULTANEOUS QUANTIFICATION OF RIFAMPICIN, ISONIAZID, PYRAZINAMIDE, AND ETHAMBUTOL HYDROCHLORIDE IN ANTI-TUBERCULOSIS 4-FDC TABLET

2015 ◽  
Vol 77 (1) ◽  
Author(s):  
Sholihul Khoiri ◽  
Sudibyo Martono ◽  
Abdul Rohman
Keyword(s):  
High Performance ◽  
Phosphate Buffer Solution ◽  
Gradient Elution ◽  
Hplc Method ◽  
Fixed Dose ◽  
Buffer Solution ◽  
Solution Ph ◽  
Combination Tablet ◽  
Simultaneous Quantification ◽  
Dose Combination

High-performance liquid chromatography (HPLC) method has been developed and validated for the simultaneous quantification of four components, namely rifampicin (RIF), isoniazid (INH), pyrazinamide (PYR), and ethambutol hydrochloride (ETM), contained in anti-tuberculosis drugs in fixed dose combination tablet (4-FDC). In order to increase the sensitivity of EMB, the pre-column derivatization technique with phenethyl isocyanate (PEIC) was carried out. The separation was accomplished using Waters Symmetry C8 (250× 4.6 mm i.d.; 5 μm) at 30oC. The mobile phase used was a mixture of acetonitrile and 20 mM phosphate buffer solution (pH 6.8) containing triethylamine and delivered at 1.5 mL/minute using gradient elution. TheUV detector was set at 210 nm. The method was validated in terms of selectivity, linearity, accuracy, precision, detection limit, quantification limit, and robustness according to International Conference on Harmanization (ICH). The optimized method is succcesfully used for quantitative analysis of RIF, INH, PYR and ETM in 4-FDC tablets. The level of these drugs in 4-FDC tablets were in accordance to that specified in Indonesian pharmacopeia.

scholarly journals Quantitation of Pregabalin by HPLC-UV Method using Ninhydrin Derivatization: Development and Validation

2020 ◽  
Vol 17 (1) ◽  
pp. 165-171
Author(s):  
Fathiy Mutalabisin ◽  
Abul Bashar Mohammed Helaluddin ◽  
Pinaki Sengupta ◽  
Farahidah Mohamed ◽  
Bappaditya Chatterjee
Keyword(s):  
High Performance ◽  
Phosphate Buffer Solution ◽  
High Performance Liquid Chromatographic ◽  
Hplc Method ◽  
Uv Detector ◽  
Buffer Solution ◽  
Solution Ph ◽  
Acid Analogue ◽  
Development And Validation ◽  
Liquid Chromatographic

Introduction: A simple and reliable high performance liquid chromatographic method has been developed for the quantitative determination of pregabalin in bulk and dosage form. Pregabalin, a γ amino butyric acid analogue, has negligible sensitivity to UV or fluorescence detection. Hence, it has been derivatized by ninhydrin to form a chromophoric complex that could be quantified by UV detection. Materials and Methods: The concentration of ninhydrin was set to 5 mg/ml and a phosphate buffer solution (pH 7.4) was used as a solvent for the reaction. The resultant complex was separated by HPLC and detected by a UV detector at 569nm wavelength. Results: The developed method showed a linear response within 50 to 600 μg/mL of pregabalin. The method was accurate with mean recovery values within 100 ± 2%. The repeatability of the method was established by intra-day and inter-day precision study. Finally, a commercial pregabalin capsule was assayed by the developed HPLC method including ninhydrin derivatization. The result of the mean assay was found to be 100.37 ±2.94 %. Conclusion: This is the first time we are reporting pregabalin analysis using ninhydrin derivatization for HPLC analysis. Therefore, the developed method can be considered as a significant improvement in pregabalin quantitation and it can be easily applied for routine quality control tests of pregabalin.

scholarly journals Development and Validation of an HPLC Method for Simultaneous Quantification of Clopidogrel Bisulfate, Its Carboxylic Acid Metabolite, and Atorvastatin in Human Plasma: Application to a Pharmacokinetic Study

2015 ◽  
Vol 2015 ◽  
pp. 1-12 ◽  
Author(s):  
Octavian Croitoru ◽  
Adela-Maria Spiridon ◽  
Ionela Belu ◽  
Adina Turcu-Ştiolică ◽  
Johny Neamţu
Keyword(s):  
Carboxylic Acid ◽  
Internal Standard ◽  
Gradient Elution ◽  
Reversed Phase ◽  
Hplc Method ◽  
Acid Metabolite ◽  
Buffer Solution ◽  
Solution Ph ◽  
Limit Of Quantification ◽  
Simultaneous Quantification

A simple, sensitive, and specific reversed phase liquid chromatographic method was developed and validated for simultaneous quantification of clopidogrel, its carboxylic acid metabolite, and atorvastatin in human serum. Plasma samples were deproteinized with acetonitrile and ibuprofen was chosen as internal standard. Chromatographic separation was performed on an BDS HypersilC18column (250 × 4.6 mm; 5 μm) via gradient elution with mobile phase consisting of 10 mM phosphoric acid (sodium) buffer solution (pH = 2.6 adjusted with 85% orthophosphoric acid) : acetonitrile : methanol with flow rate of 1 mL·min−1. Detection was achieved with PDA detector at 220 nm. The method was validated in terms of linearity, sensitivity, precision, accuracy, limit of quantification, and stability tests. Calibration curves of the analytes were found to be linear in the range of 0.008–2 μg·mL−1for clopidogrel, 0.01–4 μg·mL−1for its carboxylic acid metabolite, and 0.005–2.5 μg·mL−1for atorvastatin. The results of accuracy (as recovery) with ibuprofen as internal standard were in the range of 96–98% for clopidogrel, 94–98% for its carboxylic acid metabolite, and 90–99% for atorvastatin, respectively.

Simultaneous Determination of Oxytetracycline, Tetracycline, and Chlortetracycline in Milk by Liquid Chromatography

1989 ◽  
Vol 72 (4) ◽  
pp. 564-567 ◽  
Author(s):  
Michael H Thomas
Keyword(s):  
Bovine Milk ◽  
Phosphate Buffer Solution ◽  
Gradient Elution ◽  
Buffer Solution ◽  
Reverse Phase ◽  
Phase Gradient ◽  
Simultaneous Quantification ◽  
Molecular Weight Cutoff ◽  
Tetracycline Residues

Abstract A simple, rapid procedure is described for the simultaneous quantification of 3 tetracycline drugs in bovine milk. Samples are prepared by dilution with an EDTA/phosphate buffer solution and filtration through a molecular weight cutoff filter. Analytes are concentrated on-column using a reverse-phase gradient elution system of oxalic acid, acetonitrile, and methanol. The limits of quantitation are approximately 15-50 ng/mL and the limits of detection are 10-20 ng/ mL, depending on the compound. For oxytetracycline, over the range 50-1200 ng/mL, the average recovery and intralaboratory coefficient of variation were 97% and 4.1%, respectively. Over the same range, these parameters were, respectively, 97% and 5.0% for tetracycline, and 89% and 6.4% for chlortetracycline. The applicability of this procedure is demonstrated by separation and detection of incurred tetracycline residues in milk from treated animals.

scholarly journals Development of the simultaneous analysis of choline salicylate, lidocaine hydrochloride and preservatives in a new dental gel by HPLC method

Chemija ◽  
2021 ◽  
Vol 32 (2) ◽  
Author(s):  
Yuliia Maslii ◽  
Ivan Bezruk ◽  
Anna Materiienko ◽  
Olena Ruban ◽  
Liudas Ivanauskas ◽  
...  
Keyword(s):  
Quantitative Determination ◽  
High Performance ◽  
Limit Of Detection ◽  
Phosphate Buffer Solution ◽  
Hplc Method ◽  
Lidocaine Hydrochloride ◽  
Buffer Solution ◽  
Limit Of Quantification ◽  
Intermediate Precision

A new high-performance liquid chromatography (HPLC) method for the simultaneous quantitative determination of active pharmaceutical substances and preservatives in a new dental medication has been developed. The optimization of HPLC method parameters was done through studies of a mobile phase composition and a detection wavelength. Our developing method uses an ACE C18 column (250 × 4.6 mm, 5 µm) and a gradient mode for separation with the acetonitrile and phosphate buffer solution (adjusted to pH 3.0) as mobile phases. The flow rate is 1 ml/ min, and the detection was set at 260 nm (DAD). The method was evaluated according to the ICH guidelines and the State Pharmacopoeia of Ukraine in terms of specificity, accuracy, linearity and precision (repeatability and intermediate precision). The limit of detection and the limit of quantification were also calculated. The developed method was put in place for the analysis of a combined dental gel to a quantitative determination of the APIs (choline salicylate, lidocaine hydrochloride) and preservatives (methylparaben, propylparaben).

scholarly journals COMPATIBILITY STUDY BETWEEN PRAVASTATIN AND EZETIMIBE, AND PHARMACEUTICAL EXCIPIENTS USED IN FIXED-DOSE COMBINATION TABLET

2021 ◽  
pp. 84-89
Author(s):  
KANG MIN KIM
Keyword(s):  
High Performance ◽  
Physical Stability ◽  
Drug Formulation ◽  
Fixed Dose Combination ◽  
Fixed Dose ◽  
Compatibility Study ◽  
Scanning Calorimetry ◽  
Combination Tablet ◽  
Tg Analysis ◽  
Dose Combination

Objective: The purpose of this study was to qualitatively predict drug-excipient binding interactions for stable drug formulation of a pravastatin and ezetimibe fixed-dose combination (FDC) tablet. Methods: Drug impurity-excipient interactions under accelerated conditions (40°C/75% relative humidity) for 4 weeks were confirmed by high-performance liquid chromatography, X-ray diffraction (XRD), differential scanning calorimetry (DSC) and thermogravimetric (TG) analysis. Results: Pravastatin impurity was affected by four excipients under accelerated conditions for 4 weeks. Ezetimibe was affected by two excipients. Any other results were within the acceptance criteria. XRD analysis for physical stability revealed characteristic peaks of pravastatin and ezetimibe at a diffraction angle of 2θ (pravastatin: 4.1–24.4°, and ezetimibe: 13.62–29.59°) without a change in the crystalline form after 4 weeks. DSC and TG analysis showed evidence of stability in Alu-Alu foil. Conclusion: Thus, the tested excipients were confirmed to be compatible with pravastatin and ezetimibe and can be used in FDC bi-layer tablets.

scholarly journals ENANTIOMETRIC SEPARATION OF SITAGLIPTIN IN A FIXED DOSE COMBINATION FORMULA OF SITAGLIPTIN AND METFORMIN BY A CHIRAL LIQUID CHROMATOGRAPHIC METHOD

2019 ◽  
pp. 66-71
Author(s):  
Nagadeep Jaishetty ◽  
KAMARAJ PALANISAMY ◽  
ARTHANAREESWARI MARUTHAPILLAI
Keyword(s):  
Liquid Chromatography ◽  
Mobile Phase ◽  
Gradient Elution ◽  
Hplc Method ◽  
Fixed Dose Combination ◽  
Fixed Dose ◽  
Metformin Hydrochloride ◽  
Combination Product ◽  
Dose Combination

Objective: Sitagliptin phosphate and metformin hydrochloride tablet is an FDA approved combination product for the treatment of diabetes mellitus type 2. There are no reported evidence for estimation of undesired (S)-sitagliptin in a combination product. The objective of this study was to develop a high sensitive liquid chromatography method for the determination of (S)-enantiomer of sitagliptin phosphate in a fixed dose combination formula of metformin and sitagliptin. Methods: The proposed novel high-performance liquid chromatography (HPLC) method uses programmed gradient elution of a mixture of ethanol-diethylamine(DEA) 100:0.1 (v/v) as mobile phase-A and a mixture of methanol-water 60:40 (v/v) as mobile phase-B. The chromatographic conditions were designed to nullify the metformin interference and in which sitagliptin enantiomers elute first and followed by metformin. A satisfactory resolution (≥2.5) between (S)-sitagliptin and active form (R)-sitagliptin was achieved with gradient elution on Chiralpak IA column (5μm, 4 × 250 mm) at a flow rate of 0.5 ml/min and the detector wavelength set at 265 nm. The injection volume set as 10 µl. The developed method has been validated as per the International Conference on Harmonisation (ICH) guidelines. Results: The proposed HPLC method for determination of (S)-sitagliptin, showed good linearity in the concentration range of 0.5 µg/ml to 13.6 µg/ml and capable to quantify accurately up to the lowest level (LOQ) of 0.017%. The validated method was successfully applied to quantify the (S)-sitagliptin for different marketed formulations of sitagliptin with metformin and sitagliptin alone, and the corresponding recovery values were found to be in the range of 95.1% to 98.4%. Conclusion: The proposed validated HPLC method was found to be suitable for the quantitative determination of (S)-sitagliptin in the formulations of sitagliptin with metformin and sitagliptin alone.

Applications of Reversed-Phase High-Performance Liquid Chromatoraphy in Strains Identification of the Bean Sprout

2011 ◽  
Vol 347-353 ◽  
pp. 354-359
Author(s):  
Yong Zuo ◽  
Shuai Ju ◽  
Yang Li ◽  
Hui Xie ◽  
Li Ping Liu ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Mobile Phase ◽  
High Performance ◽  
Phosphate Buffer Solution ◽  
Reversed Phase ◽  
Qualitative And Quantitative Analysis ◽  
Buffer Solution ◽  
Solution Ph ◽  
Bean Sprout ◽  
Qualitative And Quantitative

Lactic acid which was fermented by bean sprout was determined and the lactobacillus in bean sprout was identified by High-performance Liquid Chromatoraphy(HPLC). The optimal HPLC chromatographic conditions were determined as follows:Agilent XDB-C18 chromatography column with UV detection at 210nm.The mobile phase was Phosphate buffer solution（pH=2.3）:methanol:acetonitrile（95%:2%:3%）.The flow rate was 0.5ml/min.This method is accurate,Speedily and reproducible. This method can identify the lactobacillus, and gives a qualitative and quantitative analysis of the lactic acid which was the metabolites of Yibin bean sprout.

scholarly journals Development and Validation of an HPLC Method for Simultaneous Assay of MCI and MI in Shampoos Containing Plant Extracts

2019 ◽  
Vol 2019 ◽  
pp. 1-10
Author(s):  
Le Thi Huong Hoa ◽  
Vo Tran Ngoc Hung ◽  
Do Thu Trang ◽  
Thai Nguyen Hung Thu ◽  
Dinh Chi Le
Keyword(s):  
Plant Extracts ◽  
Mobile Phase ◽  
Phosphate Buffer Solution ◽  
Hplc Method ◽  
Isopropyl Myristate ◽  
Total Flow ◽  
Buffer Solution ◽  
Solution Ph ◽  
Development And Validation

A simple, easy-to-implement HPLC method was developed and validated for simultaneous determination of two isothiazolinone preservatives, methylchloroisothiazolinone (MCI) and methylisothiazolinone (MI), in hair care shampoo containing plant extracts. In this method, shampoo samples were first dissolved in isopropyl myristate and then MCI and MI were extracted from isopropyl myristate layer by a mixture of methanol and 0.02 M phosphate buffer solution pH 3.0 (30: 70, v/v) and analyzed on an analytical biphenyl column maintained at 25°C with a mixture of methanol and water (10: 90, v/v) in isocratic elution mode as mobile phase. Total flow rate of mobile phase was maintained at 1.0 mL per minute. The UV detection was performed at 274 nm. Injection volume was 50 μl. The method was fully validated in terms of specificity, linearity, precision, accuracy, and robustness according to requirements of AOAC International and was proved as reliable and suitable for the intended application.

Development of RP-HPLC method for the analysis of levocetirizine. 2HCl and ambroxol. HCl in combination and its application

2010 ◽  
Vol 3 (1) ◽  
pp. 893-896
Author(s):  
Venisetty R K ◽  
Kamarapu S K ◽  
Vaijayanthi ◽  
Bahlul ZEA
Keyword(s):  
High Performance ◽  
Limit Of Detection ◽  
Phosphate Buffer Solution ◽  
High Performance Liquid Chromatographic ◽  
Reversed Phase ◽  
Pharmaceutical Products ◽  
Hplc Method ◽  
Buffer Solution ◽  
Rp Hplc ◽  
Limit Of Quantitation

A simple, sensitive isocratic and reproducible reversed phase High Performance Liquid Chromatographic (RP-HPLC) method was developed for the estimation of ambroxol hydrochloride (ABH) and levocetirizine dihydrochloride (LCD) in combination using PDA detector. The system consisted of RP-C18 column and the detection was performed at 230nm. The mobile phase was a mixture of acetonitrile : phosphate buffer solution (60:40) (pH  7.0) pumped at room temperature and a flow rate of 1 ml/min. ABH and LCD were eluted at 2.75 and 5.01 sec respectively. The mean absolute recoveries of ABH and LCD were about 98 % and 99 % respectively and the limit of detection of LCD and ABH in the mixture of given proportion is observed to be 0.1 µg/ml and 1.5 µg/ml and the limit of quantitation is 0.3 µg/ml and 4.5 µg/ml respectively. The calibration was linear over a concentration range of 4.5 µg/ml to 15.0 µg/ml with r2 > 0.997 for ABH and 0.3 µg/ml to 1.0 µg/ml with r2 > 0.999 for LCD. The intra (n = 5) and inter (n = 5) day assay variations in the linear range are < 4 % for ABH and < 6 % for LCD. Three pharmaceutical products containing this combination are analyzed to test the applicability of the new method. The percentage of ABH and LCD in three marketed capsule dosage form studied range from 99 to 102 % and 100 to 103 % and respectively to the claimed value.

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