scholarly journals VALIDATION METHODS FOR THE ANALYSIS OF HYDROXYPROLINE FROM COLLAGEN UNDENATURED TYPE II COLLAGEN USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY FLUORESCENCE

2018 ◽  
Vol 10 (1) ◽  
pp. 403
Author(s):  
Baitha Palanggatan Maggadani ◽  
Harmita . ◽  
Milza Lubnan

Objective: This study aimed to validate an high-performance liquid chromatography method for analyzing undenatured Type II collagen preparationsusing a fluorescence detector.Methods: Based on the optimum analysis conditions, the compound was detected at an excitation wavelength of 255 nm and an emission wavelengthof 320 nm. The optimum mobile phase was determined to be acetate (pH 4.2) and acetonitrile (60:40) with a flow rate of 1.0 ml/min. Hydroxyprolineis a compound that does not have chromophore moiety; thus, it has to be derivatized first using 9-fluorenylmetoxycarbonyl-chloride.Results: The developed method was validated with linearity and an equation of y=3,249,704 x+141,945,072, with a value of r=0.9994. The detectedrange of hydroxyproline was 4–15 ppm. The limit of detection was determined to be 0.49, with an limit of quantitation of 1.64.Conclusion: Our results indicated that the average level of hydroxyproline was 98.66%, 99.12%, and 99.85%.

2020 ◽  
Vol 11 (02) ◽  
pp. 219-223
Author(s):  
Ansari Yaasir Ahmed ◽  
Qazi Shoeb ◽  
Umme Rumana ◽  
Patel Afroza ◽  
Pathan Vahid Tajkhan ◽  
...  

The new stability-indicating high performance liquid chromatography (HPLC) method has been developed and validated with different parameters for atenolol (ATE) and nifedipine (NIFE) in the combined dosage form. The chromatographic conditions were optimized using a mobile phase of MeOH:OPA (70:30) with a flow rate of 0.7 mL/min. Column (C18) of 4.6 × 250 mm dimension was used as a stationary phase; the particle size capacity of the column was 5 μm. The detection was carried out at 233 nm. The method was validated according to ICH guidelines for linearity, precision, repeatability, the limit of detection (LoD), and limit of quantitation (LoQ). The response was found to be linear in the concentration range of 20 to 100 mcg/mL for ATE and 1 to 5 mcg/mL for NIFE. The developed method shows the minimum quantity of drugs to be identified (LoD) and minimum drug to be quantified (LoQ). The LoD and LoQ were found to be 0.1415 and 0.4289, respectively, for ATE, and 0.1834 and 0.5558, respectively, for NIFE. The method was linear, simple, precise, and accurate and, therefore, suitable for routine analysis of drugs in tablet form. The forced degradation studies were also done through the exposure of analyte solution to four different stress conditions.


2010 ◽  
Vol 7 (3) ◽  
pp. 962-966 ◽  
Author(s):  
Naveen Kumar ◽  
Nishant Verma ◽  
Omveer Songh ◽  
Naveen Joshi ◽  
Kanwar Gaurav Singh

A simple, precise, sensitive, fast and accurate high performance liquid chromatography method has been developed for the determination of atenolol using mixture of phosphate buffer and acetonitrile (53:47 v/v) as mobile phase. Buffer was prepared by mixing 0.02 M K2PO4and 0.003 M KH2PO4in equal proportion. Detection was carried out using UV detector at λmax230 nm. Column was ODS and dimensions of column was 25 mm × 4.6 mm. Atenolol was eluted out at retention time of 2.1 min. Method was validated at 1.2 mL/min flow rate. Calibration curve was linear between ranges of 40 to 200 mcg concentration. The limit of detection was calculates 120 nano gram and limit of quantitation is 510 nano gram. The relative standard deviation (RSD) of atenolol was 0.6. The percentage recovery of atenolol was 99.6%.


2014 ◽  
Vol 68 (7) ◽  
Author(s):  
Joanna Ronowicz ◽  
Bogumiła Kupcewicz ◽  
Łukasz Pałkowski ◽  
Piotr Bilski ◽  
Tomasz Siódmiak ◽  
...  

AbstractThe aim of the study was to design and validate a reversed phase high performance liquid chromatography method for the separation and quantification of two active pharmaceutical ingredients (ciprofloxacin hydrochloride, hydrocortisone) and a preservative (benzyl alcohol) in ear drops. Effective separation of the examined compounds was achieved on a GraceSmart™ RP 18 column (150 mm × 4.6 mm, 5 μm) with gradient elution and a diode array detector. The total assay run time was 25 min. Analytical method validation assays were performed. Validation parameters used for the evaluation were: specificity, linearity, trueness, precision (repeatability and reproducibility), limit of detection and limit of quantitation. Results of the validation procedure (high recoveries, good standard deviations, no interfering peaks at the retention times corresponding to the analytes) confirm that the developed chromatographic method can be applied for routine analysis of ear drops.


Author(s):  
LALITHA KV ◽  
RAVEENDRA REDDY J ◽  
DEVANNA N

Objective: This assessment depicts the strength of exhibiting reverse-phase high performance liquid chromatography (RP-HPLC) method for the estimation of torsemide in pharmaceutical estimation structures. Methods: In the present work, total protein-HPLC technique has been produced for the estimation of torsemide active pharmaceutical ingredient (API). Constrained degradation HPLC strategy was created with versatile mobile phase of methanol:water in the proportion of 90:10 v/v. The stream pace of 1 ml/min was utilized on Inertsil ODS 3V segment (250 mm×4.6 mm, 5 μm molecule size). Results: The retention time of torsemide was seen at 8.267 min, method was validated for all validation parameters as per the International Council for Harmonization guidelines. The linearity range was 10–60 μg/ml, correlation coefficient was 0.9993, and percentage relative standard deviation in the precision studies was <2%, with percentage recovery 100.56–101.03 (within acceptable range of 98–102%). The assay result was found to be 100.88% (i.e., within 95–105%), passes the specifications for robustness parameters. Limit of detection of torsemide was found to be 0.0162 μg/ml and limit of quantitation of torsemide was found to be 0.0534 μg/ml. Conclusion: The medication was exceptionally delicate to antacid pursued by at risk to corrosive, photolytic, warm, and oxidative conditions. The created and approved method showing HPLC technique is observed to be direct, exact, precise, explicit, and powerful. Henceforth, the technique can be utilized routinely for the estimation of torsemide API.


2007 ◽  
Vol 90 (2) ◽  
pp. 354-357 ◽  
Author(s):  
Mahboob Nemati ◽  
Hadi Valizadeh ◽  
Masood Ansarin ◽  
Farank Ghaderi

Abstract A column high-performance liquid chromatography (HPLC) method was developed for the determination of glucosamine in dosage forms. Glucosamine was derivatized by addition of a solution containing orthophthaldialdehyde. The HPLC separation was achieved on a Spherimage 80 ODS2 column (250 4 mm id, 5 μm particle size) using an isocratic mobile phase containing phosphate buffermethanol (90 + 10, v/v, pH 6.50) and methanoltetrahydrofuran (97 + 3, v/v) in proportions of 85 + 15 at a flow rate of 1 mL/min, followed by fluorescence detection. The method was validated for specificity, linearity, accuracy, precision, limit of detection (LOD), and limit of quantitation (LOQ). The detector response for glucosamine HCl was linear over the concentration range of 0.120 μg/mL with a correlation coefficient of 0.9980. The accuracy was between 99.4 and 100.8%. The LOD and the LOQ were 0.009 and 0.027 μg/mL, respectively. The method was applied to determination of glucosamine in solid dosage forms.


Author(s):  
Raju Chandra ◽  
Manisha Pant ◽  
Harchan Singh ◽  
Deepak Kumar ◽  
Ashwani Sanghi

A reliable and reproducible reversed-phase high performance liquid chromatography (RP-HPLC) was developed for the quantitative determination of Remipril drug content from marketed bulk tablets. The active ingredient of Remipril separation achieved with C18 column using the methanol water mobile phase in the ratio of 40:60 (v/v). The active ingredient of the drug content quantify with UV detector at 215 nm. The retention time of Remipril is 5.63 min. A good linearity relation (R2=0.999) was obtained between drug concentration and average peak areas. The limit of detection and limit of quantification of the instrument were calculated 0.03 and 0.09 µg/mL, respectively. The accuracy of the method validation was determined 102.72% by recoveries method.


2020 ◽  
Vol 103 (4) ◽  
pp. 1181-1189
Author(s):  
Xiaoyun Duan ◽  
Xiaofeng Liu ◽  
Yue Dong ◽  
Jing Yang ◽  
Jing Zhang ◽  
...  

Abstract Background Sulfonamides have been widely used in the prevention and clinical treatment of bacterial diseases in livestock and poultry. The use of sulfonamides increases the risk of veterinary drug residues in animal derived foods. The traditional reversed phase liquid chromatography methods for sulfonamides residues detection in animal derived foods have the problem of high consumption of organic solvents. Objective The aim of this study was to establish a green high-performance liquid chromatography method for the detection of sulfonamides residues in different animal-origin foods. Method The sample extraction solutions were purified by the Agela Cleanert PEP-2 cartridge and analyzed by the high-performance liquid chromatography method using ethanol as the green alternative solvent. Results The proposed method was validated in terms of linear range (20–1000 μg/kg), limit of detection (3.0–12.3 μg/kg), limit of quantitation (10–43 μg/kg), accuracy (80.7–101.3%), and repeatability and reproducibility (RSD &lt;5.9% and RSD &lt;8.5% respectively). Conclusions The proposed method is an environmentally friendly, sensitive and reliable high-performance liquid chromatography method for simultaneous determination of sulfonamide residues in animal-origin foods. Highlights In this work, we firstly developed a green high-performance liquid chromatography method for simultaneous determination of the residues of nine sulfonamides in milk and beef with ethanol as the green alternative solvent.


2011 ◽  
Vol 140 ◽  
pp. 296-301 ◽  
Author(s):  
Cai Mei Wu ◽  
Hong Min Yuan ◽  
Gang Jia ◽  
Zhi Sheng Wang ◽  
Xiu Qun Wu

A reversed high performance liquid chromatography method was developed for the quantitative determination of mimosine and 2,3-DHP in leaves ofLeucaena Leucocephala. Mimosine and 2,3DHP were extracted using 0.1N HCl.The chromatograph conditions were investigated and optimized. The optimal HPLC conditions as follows: Agilent HC-C18 column (4.6×150mm,5μm) was used at 30°C. The method used a variable wavelength UV detector at 280nm, the mobile phase consisted of 0.2 % (w/v) orthophosphoric acid and methanol, the gradient elution was adopted. The injection volume was 10μL. The linearity is favorable in the range of 1.0 to 50μg mL-1with a correlation coefficient of 0.99998 for mimosine and 0.99902 for 2,3DHP. Under the optimal conditions, the method limit of detection (LOD) of mimosine and 2,3DHP were 0.40mg/kg and 0.55mg/kg respectively. The recovery of mimosine was 87.00-94.70% with the RSD (n=5) of 2.75-3.81% in the spiked levels 0,1, 5, 20mg/g. At the same time, the recovery of 2,3DHP was 88-95.4% with the RSD (n=5) of 2.24-4.90%. The method was found to be simple, sensitive, fast and accurate, and has been applied successfully for the quantitative detection of mimosine and 2,3-DHP in leaves ofLeucaena Leucocephala, plasma and excretion of ruminant.


Author(s):  
BAITHA PALANGGATAN MAGGADANI ◽  
JIHAN YASMINA ◽  
HARMITA HARMITA

Objective: Whitening cream is a cosmetic that contains ingredients that can alleviate hyperpigmentation. Tranexamic acid (TA) is one of the potentialanti-pigmentation agents that work through inhibiting plasmin. TA is used in cosmetic formulations at a concentration of 2.5% as a whitening andmoisturizing agent. To date, research on TA in both cosmetics and other pharmaceutical products using high-performance liquid chromatography(HPLC) has not been done directly (without derivatization). Therefore, this study aimed to develop a simple and rapid analytical method for TA(without derivatization) in cosmetic cream samples using reverse-phase HPLC and water as a solvent.Methods: Optimization was conducted by evaluating several parameters that affect sample extraction, as well as composition and mobile phasetypes. The optimal method must fulfill suitability and validation requirements. The optimal method should be able to detect and quantify TA in creamsamples without derivatization.Results: The optimal analysis condition used a ultraviolet detector at a wavelength of 210 nm, acetonitrile: double-distilled water: phosphoric acid(64:34:2) as the mobile phase and a flow rate of 0.8 mL/min. The retention time of the analyte occurred in the 2nd min.Conclusion: The analytical method that met the validation requirements was characterized using parameters such as accuracy, precision, linearity,selectivity, limit, of detection, and limit of quantitation. This method is applicable for analyzing TA content in samples with a concentration of 1.02%.


2018 ◽  
Vol 10 (1) ◽  
pp. 248
Author(s):  
Baitha Palanggatan Maggadani ◽  
Harmita . ◽  
Maizura Isfadhila

Objective: The aim of this study was to produce a selective, accurate, and faster high-performance liquid chromatography (HPLC) analytical methodfor benzalkonium chloride and glutaraldehyde in disinfectants using ultraviolet (UV)-visible detection.Methods: Glutaraldehyde has no chromophore, so it was first derivatized using 2,4 dinitro phenylhydrazine. Acetonitrile:water (75:25) was used asthe mobile phase for glutaraldehyde and acetonitrile-acetate pH 4 (75:25) for benzalkonium chloride, both at a flow rate of 1.2 mL/min. The optimizedassay was validated with respect to accuracy, precision, linearity, selectivity, limit of quantitation (LOQ), and limit of detection (LOD).Results: The method was linear for benzalkonium chloride, with correlation coefficient of 0.9995, LOD of 14.55 ppm, and LOQ of 48.51 ppm. Thecorrelation coefficient for glutaraldehyde was 0.9995, with LOD of 0.49 ppm and LOQ of 1.64 ppm. Accuracy was between 98% and 102%, andprecision was below 2% for both the tests.Conclusion: The HPLC analytical method for benzalkonium chloride and glutaraldehyde in disinfectants using UV-visible detection in this researchwas successful to produce a selective, accurate, and faster method.


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