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.

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 A Simplified, Specific HPLC Method of Assaying Thiamine and Riboflavin in Mushrooms

2019 ◽  
Vol 2019 ◽  
pp. 1-8
Author(s):  
Mohammad F. Hossain ◽  
Mamoon Rashid ◽  
Rajjit Sidhu ◽  
Randy Mullins ◽  
Susan L. Mayhew
Keyword(s):  
High Performance ◽  
Limit Of Detection ◽  
Extraction Process ◽  
Reversed Phase ◽  
Hplc Method ◽  
Water Soluble ◽  
Qualitative And Quantitative Analysis ◽  
Food Industries ◽  
Qualitative And Quantitative ◽  
Rp Hplc

Mushrooms have been used as part of the average diet and as a nutraceutical for thousands of years due to their immense health benefits. The purpose of this study was to develop a simple, fast, accurate, specific, reproducible, and robust chromatographic method to identify and quantify two water-soluble vitamins: thiamine (B1) and riboflavin (B2) in mushrooms. The method employed for qualitative and quantitative analysis of these vitamins was Reversed Phase-High Performance Liquid Chromatography (RP-HPLC) equipped with Ultraviolet–Visible (UV-Vis) Detector. The extraction process involved acid hydrolysis followed by enzymatic dephosphorylation with takadiastase enzyme. Chromatographic separation was achieved with a Shimadzu prominence HPLC system using isocratic elution mode on a Waters Xterra® MS C-18 column (4.6mm × 150mm, 5 μm) integrated with a XBridge® BEH C-18 Guard column (2.1mm × 5 mm, 5 μm). The mobile phase of this study consisted of buffer and methanol in the ratio of 80:20, where the buffer contained sodium-1-hexanesulfonate, glacial acetic acid, methanol, and pH adjusted to 3.0 with diethylamine. Vitamins were detected simultaneously at their lambda max wavelengths B1: 245nm and B2: 268nm using dual-wavelength UV detection technique to get their highest response. The proposed method was found to be specific, linear R>1.0, accurate, precise (% recovery ± SD; B1:104.45±4.5 and B2: 104.88±2.04), sensitive, (limit of detection for B1 and B2 was 0.043 and 0.029 μg/mL, respectively), and robust for mushrooms analysis. No coeluting peaks were observed at the retention time of the vitamins and all the peaks were spectrally homogenous. The standard and sample solutions were found to remain stable at cold temperature for 72 hours. In summary, our data suggest that the proposed method could be used in food industries to monitor the product quality during routine quality control purposes.

scholarly journals Effect of mobile phase buffer pH on separation selectivity of some isoquinoline alkaloids in reversed-phase systems of Pressuriz

2015 ◽  
Vol 26 (3) ◽  
pp. 45-49
Author(s):  
Ewelina Kopciał ◽  
Beata Polak ◽  
Rafał Pietraś ◽  
Paulina Mączka ◽  
Tadeusz H. Dzido
Keyword(s):  
Mobile Phase ◽  
High Performance ◽  
Reversed Phase ◽  
Retardation Factor ◽  
Isoquinoline Alkaloids ◽  
Buffer Solution ◽  
Separation Selectivity ◽  
Phase Systems ◽  
Ph Range ◽  
Layer Chromatography

Separation of some isoquinoline alkaloids (narcotine, chelidonine, dihydrocodeine, cinchonine, berberine, cinchonidine, papaverine, apomorphine) has been investigated with pressurized planar electrochromatography (PPEC) and high-performance thin-layer chromatography (HPTLC) in reversed-phase systems. The mobile phase consisted of acetonitrile and aqueous buffer (disodium phosphate and citric acid). The influence of the mobile phase buffer pH on migration distance (PPEC) and retardation factor (HPTLC) of the solutes has been investigated and compared. The results show different separation selectivity in both PPEC and HPTLC systems especially at pH range of buffer solution of the mobile phase that facilitates ionization of the solutes investigated.

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 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.

Auto-oxidation of 5,6,7 ,8-tetrahydroneopterin

Pteridines ◽  
1998 ◽  
Vol 9 (1) ◽  
pp. 26-28
Author(s):  
Toshiyuki Arai ◽  
Hiroko Mori ◽  
Hisanari Ishii ◽  
Toshinori Suzuki ◽  
Shuji Kojima ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Free Radicals ◽  
High Performance ◽  
Oxygen Free Radicals ◽  
Phosphate Buffer Solution ◽  
Reversed Phase ◽  
Buffer Solution ◽  
First Time

Summary Auto-oxidation of 5,6,7,8-tetrahydroneopterin (NPH4 ) in phosphate buffer solution (PBS) was examined using reversed-phase high-performance liquid chromatography. In the chromatograms obtained every 30 min after NPH4 was dissolved in PBS, the peak of NPH4 gradually decreased and almost disappeared 90 min after the dissolution. In contrast, another peak of an unidentified substance (one of dihydroneopterins), appeared from the first time, gradually increased and became most dominant among various peaks 60 min after. The peak of 7,8-dihydroneopterin (7,8-NPH2) appeared 30 min after and gradually increased, but its height was less than that of the above-mentioned peak from first to last. These results indicate that NPH4 is readily oxidized by dissolved oxygen in PBS to produce 7,8-NPH2 and other dihydroneopterins before the reaction with oxygen free radicals and that the substances which actually serve as antioxidants in vivo are these dihydroneopterins.

scholarly journals Validation of a Simple HPLC-Based Method for Lysine Quantification for Ruminant Nutrition

Molecules ◽  
2021 ◽  
Vol 26 (14) ◽  
pp. 4173
Author(s):  
João Albuquerque ◽  
Susana Casal ◽  
Rebeca Cruz ◽  
Ingrid Van Dorpe ◽  
Margarida Rosa Garcez Maia ◽  
...  
Keyword(s):  
Amino Acids ◽  
High Performance ◽  
Phosphate Buffer Solution ◽  
Reversed Phase ◽  
Ruminant Nutrition ◽  
Buffer Solution ◽  
Chromatography Method ◽  
Nutrition Supplementation ◽  
The Matrix ◽  
Liquid Chromatography Method

Robust and selective quantification methods are required to better analyze feed supplementation effectiveness with specific amino acids. In this work, a reversed-phase high-performance liquid chromatography method with fluorescence detection is proposed and validated for lysine quantification, one of the most limiting amino acids in ruminant nutrition and essential towards milk production. To assess and widen method applicability, different matrices were considered: namely Li2CO3 buffer (the chosen standard reaction buffer), phosphate buffer solution (to mimic media in cellular studies), and rumen inoculum. The method was validated for all three matrices and found to be selective, accurate (92% ± 2%), and precise at both the inter- and intra-day levels in concentrations up to 225 µM, with detection and quantification limits lower than 1.24 and 4.14 µM, respectively. Sample stability was evaluated when stored at room temperature, 4 °C, and −20 °C, showing consistency for up to 48 h regardless of the matrix. Finally, the developed method was applied in the quantification of lysine on real samples. The results presented indicate that the proposed method can be applied towards free lysine quantification in ruminant feeding studies and potentially be of great benefit to dairy cow nutrition supplementation and optimization.

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.

scholarly journals Simultaneous Assay of Dexchlorpheniramine Maleate, Betamethasone, and Sodium Benzoate in Syrup by a Reliable and Robust HPLC Method

2019 ◽  
Vol 2019 ◽  
pp. 1-10
Author(s):  
Dinh Chi Le ◽  
Thi Duyen Ngo ◽  
Thi Huong Hoa Le
Keyword(s):  
Sodium Benzoate ◽  
Mobile Phase ◽  
Phosphate Buffer Solution ◽  
Hplc Method ◽  
Total Flow ◽  
Total Flow Rate ◽  
Buffer Solution ◽  
Solution Ph ◽  
Current Guidelines

The simultaneous determination of betamethasone, dexchlorpheniramine maleate, and sodium benzoate in pharmaceutical syrup was done by using a simple validated HPLC method. The chromatographic separation of the three analytes was done in a C18 column maintained at 25°C, using a mixture of acetonitrile and 0.02 M phosphate buffer solution pH 2.70 (35 : 65, v : v) as mobile phase. The isocratic elution was chosen with total flow rate of mobile phase maintained at 1.0 mL per minute. The analytes were detected by a UV-Vis detector set at 254 nm. Injection volume was set at 50 μl. The method was fully validated in terms of specificity, linearity, precision, accuracy, and robustness according to requirements of current guidelines and was proven to be suitable for the intended application.

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