Simultaneous determination of mycophenolic acid and its glucuronide in human plasma using a simple high-performance liquid chromatography procedure

Abstract We describe a reversed-phase HPLC method for determination of total mycophenolic acid (MPA), its free concentration (MPAf), and the glucuronide metabolite (MPAG), based on simple sample preparation and gradient elution chromatography. The compounds were quantified in parallel by absorbance at 254 nm and 215 nm in the internal standard mode. Linearity was verified up to 50 mg/L for MPA and up to 500 mg/L for MPAG (r >0.999). Detection limits at 215 and 254 nm were, respectively, 0.01 and 0.03 mg/L for MPA, and 0.03 and 0.1 mg/L for MPAG. The recovery of MPA was 95–106%;recovery of MPAG was 96–106%. The imprecision (CV) for MPA (0.2–25 mg/L) was <8.4% (254 nm) and <4.4% (215 nm) within day (n = 12) and <9.2% (254 nm) and <6.2% (215 nm) between days (n = 12). The imprecision for MPAG (10–250 mg/L) was <4.9% (254 nm) and <3.4% (215 nm) within day, and <6.1% (254 nm) and <5.9% (215 nm) between days. For quantification of MPAf, 100 μL of ultrafiltrate was applied directly to the column. The detection limit was 0.005 mg/L at 215 nm and 0.015 mg/L at 254 nm. In the range between 18–210 μg/L, the within-day CVs were <11.8% (n = 12) and the between-day CVs were <15.8% (n = 12).

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Simultaneous Determination of Eight Synthetic Permitted and Five Commonly Encountered Nonpermitted Food Colors in Various Food Matrixes by High-Performance Liquid Chromatography

Journal of AOAC International ◽

10.1093/jaoac/93.5.1503 ◽

2010 ◽

Vol 93 (5) ◽

pp. 1503-1514 ◽

Cited By ~ 9

Author(s):

Sumita Dixit ◽

Subhash K Khanna ◽

Mukul Das

Keyword(s):

Simultaneous Determination ◽

High Performance ◽

High Sensitivity ◽

Gradient Elution ◽

Reversed Phase ◽

Hplc Method ◽

Food Category ◽

Food Commodities ◽

Food Colors

Abstract A simple and sensitive HPLC method has been developed for the simultaneous determination of eight permitted food colors and five commonly encountered nonpermitted colors in various food commodities, including sugar-, fat-, and starch-based food matrixes. The method uses a specific food category-based cleanup/treatment procedure before color extraction to avoid the interference of food matrixes, and to obtain the optimal color extraction. Analysis was performed on a reversed-phase C18 -Bondapak column with ammonium acetate and acetonitrile gradient elution as the mobile phase; a programmable max-specific visible detection was used to monitor colors to obtain the higher sensitivity and expanded scope needed for multicolor blends having diverse absorption maxima. All colors showed good linearity, with regression coefficients of 0.99740.9999. The LOD and LOQ values ranged from 0.01 to 0.12 mg/L, and from 0.04 to 0.83 mg/L or mg/kg, respectively. The intraday and interday precision tests produced good RSD values, and the recoveries from different food matrixes ranged from 82 to 104%. The method offers high sensitivity for analysis of a wide variety of food matrixes containing a broad scope of multicolor blends. Two nonpermitted colors, orange II and metanil yellow, were found. Also, a number of samples contained permitted colors at levels two-to seven-fold higher than those prescribed.

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Simultaneous liquid-chromatographic determination of prednisone and prednisolone in plasma.

Clinical Chemistry ◽

10.1093/clinchem/34.9.1897 ◽

1988 ◽

Vol 34 (9) ◽

pp. 1897-1899 ◽

Cited By ~ 7

Author(s):

M H Cheng ◽

W Y Huang ◽

A I Lipsey

Keyword(s):

High Performance ◽

Internal Standard ◽

Chromatographic Determination ◽

High Performance Liquid Chromatographic ◽

Reversed Phase ◽

Hplc Method ◽

Liquid Chromatographic Determination ◽

Standard Solution ◽

Liquid Chromatographic

Abstract This high-performance liquid-chromatographic (HPLC) method for simultaneous determination of prednisone and its metabolite, prednisolone, in plasma is a modification of the method of Frey et al. (Clin Chem 1979;25:1944-7). Heparinized plasma (1.0 mL) with 0.1 mL of internal standard solution (11-deoxy-17-hydroxycorticosterone, 2 mg/L) is extracted with 7.0 mL of dichloromethane, then washed sequentially with 0.1 mol/L HCl, 0.1 mol/L NaOH, and deionized water, 2.0 mL each. The extract is evaporated and the residue reconstituted with 75 microL of mobile phase, methanol/H2O (40/60 by vol). Thirty microliters of this is injected onto a reversed-phase C6 column, which is eluted at 1.4 mL/min. Analytical recoveries of prednisone and prednisolone were 94-98% and 102-106%, respectively. Day-to-day precision (CV) was 3.8% for prednisone, 6.1% for prednisolone. We encountered no interference from the 21 other steroids and 25 drugs tested. This method is simple, accurate, and precise.

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Development and Validation of HPLC-UV Method for the Determination of a Potent Synthetic Cannabinoid THJ-2201 in Mouse Plasma and Application in a Pharmacokinetic Study

Current Pharmaceutical Analysis ◽

10.2174/1573412915666190204144843 ◽

2020 ◽

Vol 16 (4) ◽

pp. 404-411

Author(s):

Hassan Y. Aboul-Enein ◽

Gamal A.E Mostafa ◽

Haitham AlRabiah ◽

Mohammed Al-Ramadi ◽

Sabry M. Attia ◽

...

Keyword(s):

Pharmacokinetic Study ◽

High Performance ◽

Limit Of Detection ◽

Internal Standard ◽

Reversed Phase ◽

Hplc Method ◽

Synthetic Cannabinoid ◽

Mean Values ◽

Mouse Plasma

Aim: A new simple and sensitive high-Performance Liquid Chromatography (HPLC) method for the determination of a potent synthetic cannabinoid THJ-2201, has been developed and validated. Lixiviptan was used as the Internal Standard (IS). Methods: THJ-2201 and IS were extracted from mouse plasma using deproteinization procedure that uses acetonitrile followed by HPLC analysis. The separation was carried out on a reversed-phase C18 column using water and acetonitrile mixture (30:70 v/v). The flow-rate was 1.0 mL/min. Eluting of both THJ-2201 and lixivaptan was performed at 220 nm. Results: The method demonstrated linearity over a calibration range of 95 - 1500 ng/mL and the Limit of Detection (LOD) and Quantitation (LOQ) were 28 ng/mL and 91 ng/mL, respectively. The validation of the proposed method was carried out by following the US Food and Drug Administration (FDA) guidelines. Intra- and inter-day precision did not exceed 6.4%, whereas the accuracy of THJ-2201 measurements was within ±13%. Conclusion: This new method is simple and sensitive and has been applied successfully in a pharmacokinetic study of THJ-2201 in mouse plasma. The mean values of Tmax and Cmax were 0.25 h and 141.87 ± 12.11 ng/mL, respectively.

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Improved Sensitive High Performance Liquid Chromatography Assay for Glucosamine in Human and Rat Biological Samples with Fluorescence Detection

Journal of Pharmacy & Pharmaceutical Sciences ◽

10.18433/j3t01s ◽

2010 ◽

Vol 13 (2) ◽

pp. 128 ◽

Cited By ~ 13

Author(s):

Fakhreddin Jamali ◽

Alyaa Ibrahim

Keyword(s):

Fluorescence Detection ◽

Biological Samples ◽

Mobile Phase ◽

High Performance ◽

Internal Standard ◽

Reversed Phase ◽

Hplc Method ◽

Rat Urine ◽

Derivatizing Agent

Purpose. An improved HPLC method with fluorescence detection was developed and validated for determination of glucosamine in human and rat biological samples. Method. Aliquot of 0.1 mL plasma was spiked with mannosamine HCl as the internal standard (IS); proteins were precipitated with acetonitrile; the clear layer was derivatized with 9-fluorenylmethyl chloroformate (8 mM/acetonitrile) in presence of borate 0.2 M buffer at 30o C for 30 min. The excess derivatizing agent was removed with 1-aminoadamantane HCl (300 mM in acetonitrile-water 1:1). Chromatographic separation was achieved on a C18 (100mm X 4.6 mm, id 3μm) reversed phase column using 0.1% acetic acid/acetoniltrile gradient mobile phase at 1 mL/min flow rate. Glucosamine was determined in the plasma of a human and rats and also in rat urine. Results. The analytes were detected at excitation and emission wavelengths of 263 and 315 nm, respectively. The assay was linear over the range of 0.05-20 µg/mL with a typical correlation coefficient of 0.999 and intra-day and inter-day coefficient of variation of

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High Performance Liquid Chromatoqraphic Method for the Simultaneous Determination of Labetalol and Hydrochlorothiazide in Tablets and Spiked Human Plasma

Scientia Pharmaceutica ◽

10.3797/scipharm.aut-04-13 ◽

2004 ◽

Vol 72 (2) ◽

pp. 143-155 ◽

Cited By ~ 3

Author(s):

M. Sultan ◽

H. Abdine ◽

N. Zoman ◽

F. Belal

Keyword(s):

Simultaneous Determination ◽

Human Plasma ◽

High Performance ◽

Internal Standard ◽

Reversed Phase ◽

Hplc Method ◽

Spiked Human Plasma ◽

Quantitation Limit

A reversed-phase HPLC method with spectrophotometric detection was developed for the simultaneous determination of labetalol (LBT) and hydrochloro-thiazide (HCD). The chromatographic separation was performed using a Microbondapak C18 column (4.6 i.d. x 250 nm) and paracetamol as internal standard. A mobile phase consisting of 0.05 M phosphate buffer/acetonitrile of pH 4 (7:3) at a flow rate of 0.7 ml/min was used. The detection was affected spectrophotornetrically at 302 nm. The working concentration range was 0.3–10 µg/ml with detection limits of 0.05 µg/ml for both drugs. The lower quantitation limit was 0.25 µg/ml in the two cases. The method was successfully applied to tablets, the % recoveries were 99.45 ± 0.68 for LBT and 99.79 ± 0.75 for HCD. The method was extended to the in-vitro determination in spiked human plasma. The % recoveries were 91.12 ± 0.33 for LBT and 91.37 ± 0.40 for HCD. The interday and intraday precision and accuracy were evaluated in plasma by calculating the % RSD (n=5) and the % error and were found to be in the ranges of 1.18–4.1% and 0.38–0.36% for both drugs, respectively.

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Determination of Catechin and Epicatechin Content in Chocolates by High-Performance Liquid Chromatography

International Scholarly Research Notices ◽

10.1155/2014/628196 ◽

2014 ◽

Vol 2014 ◽

pp. 1-5 ◽

Cited By ~ 13

Author(s):

Raju V. S. S. Gottumukkala ◽

Nareshraju Nadimpalli ◽

Kannababu Sukala ◽

Gottumukkala V. Subbaraju

Keyword(s):

High Performance ◽

Gradient Elution ◽

High Performance Liquid Chromatographic ◽

Reversed Phase ◽

Hplc Method ◽

High Concentration ◽

Uv Visible ◽

Liquid Chromatographic ◽

Interday Precision

A simple and sensitive reversed phase high-performance liquid chromatographic (HPLC) method has been developed for the determination of catechin and epicatechin in cocoa powder and chocolates. The separation was achieved on a reversed phase C 18 column (TARGA) 5 μm by gradient elution with a flow rate of 1.0 mL/minute with an operating temperature of 30°C and detection with a UV-Visible detector was at 280 nm. The method was validated for linearity, precision, intra- and interday precision, and accuracy. The developed method is successfully applied for the determination of catechin and epicatechin content in chocolates. The Godiva brand chocolate contains high concentration of epicatechin.

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Developing a High-performance Liquid Chromatography Method for Simultaneous Determination of Loratadine and its Metabolite Desloratadine in Human Plasma

Current Drug Metabolism ◽

10.2174/1389200220666191125095648 ◽

2020 ◽

Vol 20 (13) ◽

pp. 1053-1059

Author(s):

Mahmoud M. Sebaiy ◽

Noha I. Ziedan

Keyword(s):

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

Human Plasma ◽

High Performance ◽

Allergic Diseases ◽

Reversed Phase ◽

Hplc Method ◽

H1 Receptor ◽

Chromatography Method

Background: Allergic diseases are considered as the major burden on public health with increased prevalence globally. Histamine H1-receptor antagonists are the foremost commonly used drugs in the treatment of allergic disorders. The target drug in this study, loratadine, belongs to this class of drugs and its biometabolite desloratadine which is also a non-sedating H1 receptor antagonist with anti-histaminic activity being 2.5 to 4 times greater than loratadine. This study aimed to develop and validate a novel isocratic Reversed-phase High-Performance Liquid Chromatography (RP-HPLC) method for rapid and simultaneous separation and determination of loratadine and its metabolite, desloratadine in human plasma. Methods: The drug extraction method from plasma was based on protein precipitation technique. The separation was carried out on a Thermo Scientific BDS Hypersil C18 column (5μm, 250 x 4.60 mm) in a mobile phase of MeOH: 0.025M KH2PO4 adjusted to pH 3.50 using orthophosphoric acid (85: 15, v/v) at an ambient temperature. The flow rate was maintained at 1 mL/min and maximum absorption was measured using the PDA detector at 248 nm. Results: The retention times of loratadine and desloratadine in plasma samples were recorded to be 4.10 and 5.08 minutes, respectively, indicating a short analysis time. Limits of detection were found to be 1.80 and 1.97 ng/mL for loratadine and desloratadine, respectively, showing a high degree of sensitivity of the method. The method was then validated according to FDA guidelines for the determination of the two analytes in human plasma. Conclusion: The results obtained indicate that the proposed method is rapid, sensitive in the nanogram range, accurate, selective, robust and reproducible compared to other reported methods.

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Development of an HPLC method for measuring orally administered 1-substituted 2-alkyl-3-hydroxypyrid-4-one iron chelators in biological fluids

Clinical Chemistry ◽

10.1093/clinchem/36.1.5 ◽

1990 ◽

Vol 36 (1) ◽

pp. 5-8 ◽

Cited By ~ 17

Author(s):

J G Goddard ◽

G J Kontoghiorghes

Keyword(s):

High Performance ◽

Biological Fluids ◽

Internal Standard ◽

High Performance Liquid Chromatographic ◽

Reversed Phase ◽

Hplc Method ◽

Iron Chelators ◽

Serum Samples ◽

Thalassemic Patient ◽

Serum And Urine

Abstract "High-performance" liquid-chromatographic (HPLC) methods have been developed for identifying 1-substituted 2-alkyl-3-hydroxypyrid-4-one iron chelators in serum and urine. Ion pairing with heptane- or octanesulfonic acid in pH 2.0-2.2 phosphate buffer and reversed-phase chromatography were required to separate these compounds from endogenous compounds in both biological fluids. In both the 2-methyl and 2-ethyl series of 1-substituted compounds (H, methyl, ethyl, or propyl) the elution times increased in accordance with the n-octanol/water partition coefficients (propyl greater than ethyl greater than H greater than methyl). Urine samples were filtered (0.4 microns pore size) and injected either undiluted or after dilution with elution buffer. After the addition of internal standard, the plasma or serum samples were deproteinized by treatment with HCIO4, 0.5 mol/L, centrifuged, and the supernates were injected directly onto the HPLC. Using these procedures, we could identify 1,2-dimethyl-3-hydroxypyrid-4-one (L1) in the serum and urine of a thalassemic patient who had received a 3-g dose of the drug and in the urine of other patients who had received the same dose. One or more possible metabolites were also observed in the chromatograms of both urine and serum. The 24-h urinary output of L1 (0.22-2.37 g) and iron (10.6-71.5 mg) varied but there was no correlation between the two with respect to quantity or concentration. Instead, urinary iron output was higher in patients with a greater number of transfused units of erythrocytes. This is the first study in humans to show that L1 is absorbed from the gut, enters the circulation, and is excreted in the urine.

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Determination of Ten Corticosteroids in Illegal Cosmetic Products by a Simple, Rapid, and High-Performance LC-MS/MS Method

International Journal of Analytical Chemistry ◽

10.1155/2017/3531649 ◽

2017 ◽

Vol 2017 ◽

pp. 1-12 ◽

Cited By ~ 7

Author(s):

Vita Giaccone ◽

Giuseppe Polizzotto ◽

Andrea Macaluso ◽

Gaetano Cammilleri ◽

Vincenzo Ferrantelli

Keyword(s):

High Performance ◽

Skin Diseases ◽

Gradient Elution ◽

Reversed Phase ◽

Cosmetic Products ◽

Chromatography Method ◽

Esi Ms ◽

Negative Ionization Mode ◽

Negative Ionization

The aim of our present work was the development of a rapid high-performance liquid chromatography method with electrospray ionization and tandem mass spectrometry detection (LC-ESI-MS/MS) for the determination of several corticosteroids in cosmetic products. Corticosteroids are suspected to be illegally added in cosmetic preparations in order to enhance the curative effect against some skin diseases. Sample preparation step consists in a single extraction with acetonitrile followed by centrifugation and filtration. The compounds were separated by reversed-phase chromatography with water and acetonitrile (both with 0.1% formic acid) gradient elution and detected by ESI-MS positive and negative ionization mode. The method was validated at the validation level of 0.1 mg kg−1. Linearity was studied in the 5–250 μg L−1 range and linear coefficients (r2) were all over 0.99. The accuracy and precision of the method were satisfactory. The LOD ranged from 0.085 to 0.109 mg kg−1 and the LOQ from 0.102 to 0.121 mg kg−1. Mean recoveries for all the analytes were within the range 91.9–99.2%. The developed method is sensitive and useful for detection, quantification, and confirmation of these corticosteroids in cosmetic preparations and can be applied in the analysis of the suspected samples under investigation.

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Simultaneous Determination of Preservatives (Methyl Paraben and Propyl Paraben) in Sucralfate Suspension Using High Performance Liquid Chromatography

E-Journal of Chemistry ◽

10.1155/2011/360431 ◽

2011 ◽

Vol 8 (1) ◽

pp. 340-346 ◽

Cited By ~ 5

Author(s):

Rajesh M. Kashid ◽

Santosh G. Singh ◽

Shrawan Singh

Keyword(s):

High Performance Liquid Chromatography ◽

Simultaneous Determination ◽

Mobile Phase ◽

High Performance ◽

Reversed Phase ◽

Hplc Method ◽

Reversed Phase Hplc ◽

Methyl Paraben ◽

Propyl Paraben

A reversed phase HPLC method that allows the separation and simultaneous determination of the preservatives methyl paraben (M.P.) and propyl paraben (P.P.) is described. The separations were effected by using an initial mobile phase of water: acetonitrile (50:50) on Inertsil C18 to elute P.P. and M.P. The detector wavelength was set at 205 nm. Under these conditions, separation of the two components was achieved in less than 10 min. Analytical characteristics of the separation such as precision, specificity, linear range and reproducibility were evaluated. The developed method was applied for the determination of preservative M.P. and P.P. at concentration of 0.01 mg/mL and 0.1 mg/mL respectively. The method was successfully used for determining both compounds in sucralfate suspension.

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