scholarly journals An Efficient Heparin Affinity Column Purification Method Coupled with Ultraperformance Liquid Chromatography for the Quantification of Native Lactoferrin in Breast Milk

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Huizhi Yuan ◽  
Na Li ◽  
Yiping Xun ◽  
Lin Wang ◽  
Xiaoying Feng ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Breast Milk ◽  
Human Milk ◽  
Limit Of Detection ◽  
Affinity Column ◽  
Linear Calibration ◽  
Step Method ◽  
Purification Method ◽  
Limit Of Quantitation ◽  
Heparin Affinity

Lactoferrin (LF) is a bioactive multifunctional protein and found in the highest amounts in human milk. Several methods can be used to quantify LF. However, quantification of native LF has garnered relatively little interest to date. This study aimed to develop a novel efficient two-step method for quantifying native LF in breast milk. During the analysis, LF was first extracted with phosphate buffer (pH 5.0), purified using a heparin affinity column. Subsequently, LF was detected using ultraperformance liquid chromatography (UPLC) at a wavelength of 201 nm. A linear calibration curve was obtained in the range of 5–200 mg/L. The limit of detection and limit of quantitation were 1 mg/L and 5 mg/L, respectively, indicating that the validated method could be employed to quantify LF in breast milk. Compared with previous HPLC methods, this method demonstrated several remarkable advantages, including simple operation, low-cost detection, and high accuracy. Hence, the results demonstrate an efficient method that can be employed commercially to purify and analyze LF in human milk samples.

scholarly journals Rapid detection of dimethyl yellow dye in curry by liquid chromatography-electrospray-tandem mass spectrometry

2010 ◽  
Vol 28 (No. 5) ◽  
pp. 427-432 ◽  
Author(s):  
F. Tateo ◽  
M. Bononi ◽  
F. Gallone
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Tandem Mass Spectrometry ◽  
Limit Of Detection ◽  
Tandem Mass ◽  
Mass Spectral ◽  
Limit Of Quantitation ◽  
Liquid Chromatography Tandem Mass ◽  
High Degree ◽  
Positive Ion

An accurate and rapid method, was devised for the identification and quantitation of dimethyl yellow dye in curry, based on liquid chromatography-tandem mass spectrometry interfaced with electrospray. Mass spectral acquisition was done in positive ion mode applying two fragmentation transitions to provide a high degree of selectivity. The extraction system provided a very high recovery (100.0% to 105.8%) and good results were obtained for the limit of detection (5 μg/kg) and limit of quantitation (16 μg/kg). The applicability of the method to identifing and quantifing the unauthorised dimethyl yellow dye in curry was demonstrated.

scholarly journals Determination of Tianeptine in Tablets by High-Performance Liquid Chromatography with Fluorescence Detection

2007 ◽  
Vol 90 (3) ◽  
pp. 720-724
Author(s):  
Sevgi Tatar Ulu
Keyword(s):  
Lower Limit ◽  
High Performance ◽  
Limit Of Detection ◽  
Internal Standard ◽  
Linear Calibration ◽  
Fluorescence Detector ◽  
Relative Standard ◽  
Validation Parameters ◽  
Limit Of Quantitation

Abstract A sensitive and selective high-performance liquid chromatographic method has been developed for the determination of tianeptine (Tia) in tablets. The method is based on derivatization of Tia with 4-chloro-7-nitrobenzofurazan (NBD-Cl). A mobile phase consisting of acetonitrile10 mM orthophosphoric acid (pH 2.5; 77 + 23) was used at a flow rate of 1 mL/min on a C18 column. The Tia-NBD derivative was monitored using a fluorescence detector, with emission set at 520 nm and excitation at 458 nm. Gabapentin was selected as an internal standard. Linear calibration graphs were obtained in the concentration range of 45300 ng/mL. The lower limit of detection (LOD) was 10 ng/mL at a signal-to-noise ratio of 4. The lower limit of quantitation (LOQ) was 45 ng/mL. The relative standard values for intra- and interday precision were <0.46 and <0.57%, respectively. The recovery of the drug samples ranged between 98.89 and 99.85%. No chromatographic interference from the tablet excipients was found. The proposed method was validated in terms of precision, robustness, recovery, LOD, and LOQ. All the validation parameters were within the acceptance range. The proposed method was applied for the determination of Tia in commercially available tablets. The results were compared with those obtained by an ultraviolet spectrophotometric method using t- and F-tests.

scholarly journals Determination of 7B3 Residues in Cotton Plants by Liquid Chromatography/Mass Spectrometry

2009 ◽  
Vol 92 (1) ◽  
pp. 302-306 ◽  
Author(s):  
Xiao-Jing Yan ◽  
Xiao-Mei Liang ◽  
Yan-Jun Xu ◽  
Shu-Hui Jin ◽  
Dao-Quan Wang
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Limit Of Detection ◽  
Reversed Phase ◽  
Liquid Chromatography Mass Spectrometry ◽  
Chromatography Mass Spectrometry ◽  
Relative Standard ◽  
Limit Of Quantitation ◽  
Cotton Plants

Abstract A method was developed for the determination of 7B3 (12-propyloxyimino-1,15-pentadecanlactam), a novel macrolactam fungicide, by liquid chromatography/mass spectrometry (LC/MS) with positive electrospray ionization (ESI+). The method used a reversed-phase C18 column and acetonitrilewater (60 + 40, v/v) mobile phase. The quick, easy, cheap, effective, rugged, and safe method was used for extraction of 7B3 from cotton plants, which involved the extraction of 10 g homogenized sample with 10 mL acetonitrile, followed by the addition of 4 g anhydrous MgSO4 and 1.0 g NaCl. After centrifugation, 1 mL of the buffered acetonitrile extract was transferred into a tube containing 50 mg primary secondary amine sorbent and 100 mg anhydrous MgSO4. After shaking and centrifugation, the final extract was transferred to an autosampler vial for concurrent analysis by LC/MS. The results of 7B3 determined by LC/MS in the selective ion monitoring mode were linear, and the matrix effect of the method was evaluated. The average recoveries of 7B3 fortified at different levels were within 84.1100.2, and the relative standard deviations were <7.5 for all samples analyzed. The method limit of detection and the limit of quantitation values were 0.03 and 0.1 mg/kg, respectively. The proposed method was successfully applied to determine 7B3 residues in practical samples. This method is sensitive, accurate, reliable, simple, and safe.

scholarly journals Rapid Determination of Lysine in Biological Samples by Isocratic Liquid Chromatography

1999 ◽  
Vol 82 (4) ◽  
pp. 809-813 ◽  
Author(s):  
Mamun M Or-Rashid ◽  
Ryoji Onodera ◽  
Shaila Wadud ◽  
Mohamed-Emad A Nasser ◽  
Mohammad R Amin
Keyword(s):  
Liquid Chromatography ◽  
Biological Samples ◽  
Limit Of Detection ◽  
Rapid Determination ◽  
Rumen Fluid ◽  
Uv Detector ◽  
Rumen Microorganisms ◽  
Microbial Nitrogen ◽  
Limit Of Quantitation

Abstract A simple, rapid, and sensitive method was developed for detection and quantitation of lysine (Lys) in various biological samples by isocratic liquid chromatography (LC). Samples containing Lys and other amino acids were derivatized with 9-fluorenylmethyl chloroformate (FMOC-CI). The mobile phase used for isocratic elution was 50 mmol/L sodium acetate buffer (pH 4.20)-acetonitrile (43 + 57, v/v). Lys was detected with a UV detector at 265 nm. The derivatized Lys eluted from a LiChrospher 100 RP-18 (150× 4.0 mm id) column at a retention time of 5.6 min. The limit of detection was 0.73 μmol/L (signal-to-noise [S/N] ratio, 3:1), and the limit of quantitation was 2.37 μmol/L (S/N ratio, 10:1). Lys recoveries from fortified biological samples were >97.5%. Average Lys contents found in rumen fluid samples collected before the morning feeding and at 2.0,4.0, and 6.0 h after feeding were 4.26,3.34,3.58, and 3.82 μmol/L, respectively. The hydrolysate of a sample of mixed rumen microorganisms collected before the morning feeding was determined to contain 1.372 μmol/mg microbial nitrogen in the form of Lys. The Lys concentrations of human plasma, goat plasma, human urine, and goat urine were 140.0, 102.0,58.0, and 32.0 μmol/L, respectively.

scholarly journals DEVELOPMENT OF A DIRECT METHOD OF ANALYZING TRANEXAMIC ACID LEVELS IN WHITENING CREAM USING REVERSED PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

2020 ◽  
pp. 88-92
Author(s):  
BAITHA PALANGGATAN MAGGADANI ◽  
JIHAN YASMINA ◽  
HARMITA HARMITA
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Tranexamic Acid ◽  
Analytical Method ◽  
High Performance ◽  
Limit Of Detection ◽  
Direct Method ◽  
Pharmaceutical Products ◽  
Optimal Method ◽  
Limit Of Quantitation

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%.

scholarly journals A Validated Method for Detection and Quantitation of 2’Fucosyllactose in Infant Formula Matrices

2019 ◽  
Vol 7 (1) ◽  
Author(s):  
Pedro A. Prieto ◽  
Michael B. Miklus ◽  
Cynthia M. Barber ◽  
Steven M. Tennyson
Keyword(s):  
Human Milk ◽  
Infant Formula ◽  
Limit Of Detection ◽  
Soluble Carbohydrates ◽  
Infant Formulas ◽  
Chromatography Method ◽  
Elution Time ◽  
Potential Health ◽  
Main Challenge ◽  
Limit Of Quantitation

Analytical methods to assess the content of free carbohydrates in solution range from simple tests of reductive power to combinations of chromatography and mass spectrometry. Soluble carbohydrates such as lactose, maltose, fructooligosaccharides, and galactooligosaccharides are commonly found in infant formulas either as sources of energy or soluble fibers. On the other hand, a rich repertoire of lactose-based carbohydrates occurs naturally in human milk.  The advent of novel biosynthesis technologies resulted in the availability of human milk oligosaccharide structures that are being used as ingredients in infant formulas.   Notably, 2’Fucosyllactose has been tested in preclinical and clinical studies to determine its safety and to explore its potential health benefits in the context of pediatric nutrition. Several chromatographic methods for the analysis of human milk oligosaccharides have been published and, the main challenge associated with 2’Fucosyllactose quantitation has been to improve its resolution from lactose, which is present at concentrations around 70 g/l in both, infant formula and human milk. We developed a high-performance anion exchange chromatography method to detect and quantify 2’ Fucosyllactose in the presence of lactose by expanding the elution time between these saccharides. We validated the analytical procedure which behaved linearly (average R=0.99951) at concentrations as low as 1.75 µg/ml (limit of quantitation) with an average limit of detection of 0.43 µg/ml.

scholarly journals Development and Validation of Stability-Indicating Reverse Phase-High Performance Liquid Chromatography Method for Simultaneous Determination of Atenolol and Nifedipine in Bulk and Tablet Dosage Form

2020 ◽  
Vol 11 (02) ◽  
pp. 219-223
Author(s):  
Ansari Yaasir Ahmed ◽  
Qazi Shoeb ◽  
Umme Rumana ◽  
Patel Afroza ◽  
Pathan Vahid Tajkhan ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Dosage Form ◽  
Limit Of Detection ◽  
Hplc Method ◽  
Forced Degradation ◽  
Chromatography Method ◽  
Stability Indicating ◽  
Limit Of Quantitation

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.

scholarly journals Quantification of Lappaconitine in Mouse Blood by UPLC-MS/MS and Its Application to a Pharmacokinetic Study

2019 ◽  
Vol 2019 ◽  
pp. 1-6
Author(s):  
Fan Chen ◽  
Xiuwei Shen ◽  
Peng Huang ◽  
Huiyan Fu ◽  
Yue Jin ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Multiple Reaction Monitoring ◽  
Ultrahigh Pressure ◽  
Absolute Bioavailability ◽  
Quantitative Detection ◽  
Intragastric Administration ◽  
Linear Calibration ◽  
Ionization Source ◽  
Mouse Blood ◽  
Limit Of Quantitation

Lappaconitine is extracted from Aconitum sinomontanum Nakai, which belongs to the Ranunculaceae. Lappaconitine is as a diterpenoid alkaloid used as a nonaddictive analgesic. To assure the rational use of the drug, ultrahigh-pressure liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was conducted to determine lappaconitine in mouse blood and its application to pharmacokinetics. In this study, khasianine was used as internet standard (IS). A UPLC BEH C18 column was used for chromatographic separation and the mobile phase consisted of acetonitrile and 10 mmol/L ammonium acetate (0.1% formic acid). The flow rate of was 0.4 mL/min. Quantitative detection was performed in a multiple reaction monitoring (MRM) mode using an electrospray ionization source in positive mode. Twenty-four mice were randomly divided into four groups, three of which received 2, 4, and 8 mg/kg lappaconitine by intragastric administration, while the other group received 1 mg/kg lappaconitine by intravenous administration. After 0.0833, 0.5, 1, 1.5, 2, 3, 4, and 8 h, blood samples were collected and acetonitrile was used for protein precipitation. A linear calibration relationship (R2 = 0.9979) in the range of 0.1-500 ng/mL in mouse blood indicated good results. The lower limit of quantitation was 0.1 ng/mL and the limit of detection was 0.04 ng/mL. The intra-day and inter-day precision were below 13% and 14%, respectively. The accuracy was 90.1-107.2%, and the recovery exceeded 81.1%. The matrix effect ranged between 102.1 and 108.8%. The absolute bioavailability of lappaconitine was 2.0%. UPLC-MS/MS achieved high sensitivity, speed, and selectivity. Methodological verification indicated this method as suitable for determination of lappaconitine in mouse blood.

scholarly journals HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY ANALYTICAL METHOD VALIDATION FOR GLUTARALDEHYDE AND BENZALKONIUM CHLORIDE IN DISINFECTANTS

2018 ◽  
Vol 10 (1) ◽  
pp. 248
Author(s):  
Baitha Palanggatan Maggadani ◽  
Harmita . ◽  
Maizura Isfadhila
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Analytical Method ◽  
Benzalkonium Chloride ◽  
Mobile Phase ◽  
High Performance ◽  
Limit Of Detection ◽  
Analytical Method Validation ◽  
Limit Of Quantitation ◽  
Uv Visible

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.

scholarly journals Determination of Cyclamate in Foods by Ultraperformance Liquid Chromatography/Tandem Mass Spectrometry

2008 ◽  
Vol 91 (5) ◽  
pp. 1095-1102 ◽  
Author(s):  
Robert Sheridan ◽  
Thomas King
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Sample Preparation ◽  
Tandem Mass Spectrometry ◽  
Limit Of Detection ◽  
Tandem Mass ◽  
Chromatography Tandem Mass Spectrometry ◽  
Limit Of Quantitation ◽  
Liquid Chromatography Tandem Mass

Abstract A highly sensitive and selective method that requires minimal sample preparation was developed for the confirmation and quantitation of cyclamate in a variety of foods by high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS). Sample preparation consisted of homogenization followed by extraction and dilution of cyclamate with water. HPLC separation was achieved using a bridged ethyl hybrid C18 high-pressure column with a mobile phase consisting of 0.15 acetic acid and methanol. Under electrospray ionization negative conditions, quantitation was achieved by monitoring the fragment m/z 79.7 while also collecting parent ion m/z 177.9. Two food matrixes, diet soda and jelly, were subjected to a validation procedure in order to evaluate the applicability of the method. The cyclamate limit of detection for both matrixes was determined to be 0.050 g/g with a limit of quantitation of 0.150 g/g. The correlation coefficient of the calibration curves was >0.9998 from 0.0005 to 0.100 g/mL. The method has been used for the determination of cyclamate in several foods and the results are presented.

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