Specific 3H radioimmunoassay with a monoclonal antibody for monitoring cyclosporine in blood.

1988 ◽  
Vol 34 (2) ◽  
pp. 257-260 ◽  
Author(s):  
P E Ball ◽  
H Munzer ◽  
H P Keller ◽  
E Abisch ◽  
J Rosenthaler
Keyword(s):  
Monoclonal Antibody ◽  
Bone Marrow ◽  
High Performance Liquid Chromatography ◽  
High Performance ◽  
Limit Of Detection ◽  
Parent Drug ◽  
Sample Volume ◽  
Specific Radioimmunoassay ◽  
The Mean ◽  
Blood Specimens

Abstract A specific radioimmunoassay involving a mouse monoclonal antibody to cyclosporine has been developed for monitoring the parent drug in blood. Pretreatment with methanol removes cyclosporine from the erythrocytes. The limit of detection is about 12 micrograms/L, sample volume is 50 microL of blood, and within- and between-assay CVs are less than 7%. Assay results correlated well with those obtained by "high-performance" liquid chromatography (HPLC) for liver (n = 42), for heart (n = 64), for bone-marrow (n = 36), and for kidney (n = 140). For blood specimens obtained from patients treated with cyclosporine postoperatively for as long as 65 months, the mean RIA/HPLC ratio in all with transplant indications was close to 1. Therefore, the specific radioimmunoassay apparently can be used instead of HPLC to measure the parent drug in blood.

Determination of cyclosporine in plasma: specific radioimmunoassay with a monoclonal antibody and liquid chromatography compared.

1989 ◽  
Vol 35 (4) ◽  
pp. 608-611 ◽  
Author(s):  
L Vernillet ◽  
H P Keller ◽  
J F Le Bigot ◽  
H Humbert
Keyword(s):  
Monoclonal Antibody ◽  
Bone Marrow ◽  
Liquid Chromatography ◽  
Drug Monitoring ◽  
Limit Of Detection ◽  
Parent Drug ◽  
Hplc Method ◽  
Monitoring Period ◽  
Specific Radioimmunoassay

Abstract A radioimmunoassay of cyclosporine (Sandimmune) involving use of a mouse monoclonal antibody was tested to monitor specifically the parent drug in plasma. The cyclosporine concentrations obtained by RIA were compared with those obtained by the HPLC method. For the RIA method, the within- and between-assay CVs are less than 6%, the limit of detection is about 10 micrograms/L for a 50-microL sample of plasma. For the HPLC method, the within- and between-assay CVs are less than 20%, the limit of detection is about 15 micrograms/L for a 1-mL sample of plasma. The concentrations by RIA correlated well with those by HPLC in samples from patients receiving bone-marrow (n = 39), heart (n = 52), or liver (n = 51) transplants. In all indications, the ratio of values by RIA to those by HPLC for these samples remained stable and close to 1 during the drug-monitoring period, i.e., for up to 202 days. Therefore, the specific RIA can be used instead of HPLC to measure the parent drug in plasma.

An evaluation of the Abbott IMx sirolimus assay in relation to a high-performance liquid chromatography-ultraviolet method

2005 ◽  
Vol 42 (5) ◽  
pp. 394-397 ◽  
Author(s):  
Roger N Johnson ◽  
Russell Sargon ◽  
Gerald Woollard ◽  
James Davidson
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Immunosuppressive Agent ◽  
Hplc Method ◽  
Transplant Patients ◽  
Coefficients Of Variation ◽  
Microparticle Enzyme Immunoassay ◽  
The Mean ◽  
Blood Specimens

Background: Sirolimus is an immunosuppressive agent used after organ transplantation. Monitoring its concentration is clinically relevant, but current methods of measurement using high-performance liquid chromatography (HPLC) are demanding. An immunoassay that uses readily available equipment offers a more convenient alternative. Methods: Blood specimens from transplant patients were assayed for sirolimus by an existing HPLC method with ultraviolet (UV) detection and by the Abbott microparticle enzyme immunoassay (MEIA). Within-run and between-day precision was assessed with duplicate analyses. Accuracy was assessed by comparison of results from the MEIA and HPLC assays, and from recovery experiments. Specificity was tested in specimens from patients taking alternative immunosuppressants. Results: Using specimens with sirolimus concentrations between 3.3 and 39.3 µg/L, within-batch and between-batch coefficients of variation (CVs) were 6.8% and 6.3%, respectively. Using control specimens with mean concentrations of 4.1, 12.1 and 25.4 µg/L, the respective CVs were 8.6%, 3.1% and 3.8 %. Using a Passing-Bablok regression technique, the regression equation was MEIA = 14; 1.26 x HPLC-1.97; the mean bias was 0.96 µg/L. Specimens from patients on cyclosporin and tacrolimus as the sole immunosuppressant had apparent sirolimus concentrations of <1 µg/L. Conclusion: The MEIA assay for sirolimus provides clinically useful information within an accessible format.

Development and Validation of A Reversed Phase-High Performance Liquid Chromatography Method for the Quantitative Estimation of Ramipril Drug Content from Formulated Dosage Form

2016 ◽  
Vol 6 (3) ◽  
Author(s):  
Raju Chandra ◽  
Manisha Pant ◽  
Harchan Singh ◽  
Deepak Kumar ◽  
Ashwani Sanghi
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Active Ingredient ◽  
High Performance ◽  
Limit Of Detection ◽  
Quantitative Estimation ◽  
Reversed Phase ◽  
Uv Detector ◽  
Chromatography Method ◽  
Drug Content

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.

High performance liquid chromatography (HPLC) method development and validation for the determination of flunixin: Animal plasma based study

2021 ◽  
pp. 1-11
Author(s):  
Sultan M. Alshahrani ◽  
John Mark Christensen
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Method Development ◽  
Limit Of Detection ◽  
Hplc Method ◽  
Limit Of Quantification ◽  
Mobile Phases ◽  
Hplc Method Development ◽  
Development And Validation

This study was designed to develop and validate a simple and efficient high performance liquid chromatography (HPLC) method to determine flunixin concentrations in Asian elephant’s (Elephas maximus) plasma. Flunixin was administered orally at a dose of 0.8 mg/kg, and blood samples were collected. Flunixin extraction was performed by adding an equal amount of acetonitrile to plasma and centrifuging at 4500 rpm for 25 minutes. The supernatant was removed, and flunixin was analyzed using HPLC-UV detection. Two methods were developed and tested utilizing two different mobile phases either with or without adding methanol (ACN: H2O vs. ACN: H2O: MeOH). Both methods showed excellent linearity and reproducibility. The limit of detection was 0.05 ug/ml and limit of quantification was 0.1 ug/ml. the efficiency of flunixin recovery was maximized by the addition of methanol to mobile phase (ACN: H2O: MeOH as 50:30:20) at 95% in comparison to 23% without methanol. In conclusion, adding methanol to HPLC methods for extraction of flunixin from elephants’ plasma yielded higher recovery rate than without methanol.

Occurrence of Aflatoxin M1 in Cheeses from the South of Spain

1996 ◽  
Vol 59 (8) ◽  
pp. 898-900 ◽  
Author(s):  
Mª JOSÉ BARRIOS ◽  
Mª JESÚS GUALDA ◽  
J. M. CABANAS ◽  
L. M. MEDINA ◽  
R. JORDANO
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Aflatoxin M1 ◽  
The South ◽  
Different Types ◽  
The Mean

Thirty-five samples of commercial cheeses, 9 fresh, 9 semicured or semiripened and 17 ripened made with different types of milk (cow, ewe, goat and mixtures of milk of various species) produced in the South of Spain were analyzed for the presence of aflatoxin M1 (AFM1) by high-performance liquid chromatography, In 16 of the 35 samples (45.71%) the presence of AFM1 was detected in concentrations ranging between 20 and 200 ng/g of cheese, In the positive cases, the mean levels of AFM1 were 105.33 ng/g in ripened cheeses, 73.80 ng/g in semiripened cheeses and 42.60 ng/g in fresh cheeses.

scholarly journals Development and validation of reversed phase high performance liquid chromatography (RP-HPLC) for quantification of captopril in rabbit plasma

2020 ◽  
Author(s):  
Muhammad Fawad Rasool ◽  
Umbreen Fatima Qureshi ◽  
Nazar Muhammad Ranjha ◽  
Imran Imran ◽  
Mouqadus Un Nisa ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Mobile Phase ◽  
High Performance ◽  
Limit Of Detection ◽  
Reversed Phase ◽  
Rabbit Plasma ◽  
Rp Hplc ◽  
Relative Standard

AbstractTh accurate rapid, simple and selective reversed phase high performance liquid chromatography (RP-HPLC) has been established and validated for the determination of captopril (CAP). Chromatographic separation was accomplished using prepacked ODSI C18 column (250 mm × 4.6 mm with 5 μm particle size) in isocratic mode, with mobile phase consisting of water: acetonitrile (60:40 v/v), pH adjusted to 2.5 by using 85% orthophosphoric acid at a flow rate of 1 mL/min and UV detection was performed at 203 nm. RP-HPLC method used for the analysis of CAP in mobile phase and rabbit plasma was established and validated as per ICH-guidelines. It was carried out on a well-defined chromatographic peak of CAP was established with a retention time of 4.9 min and tailing factor of 1.871. The liquid–liquid extraction method was used for extraction of CAP from the plasma. Excellent linearity (R2 = 0.999) was shown over range 3.125–100 µg/mL with mean percentage recoveries ranges from 97 to 100.6%. Parameters of precision and accuracy of the developed method meet the established criteria. Intra and inter-day precision (% relative standard deviation) study was also performed which was less than 2% which indicate good reproducibility of the method. The limit of detection (LOD) and quantification for the CAP in plasma were 3.10 and 9.13 ng/mL respectively. The method was suitably validated and successfully applied to the determination of CAP in rabbit plasma samples.

scholarly journals Changes in the contents of kaempherol, quercetin and L-ascorbic acid in sea buckthorn berries during maturation

2000 ◽  
Vol 9 (1) ◽  
pp. 17-22 ◽  
Author(s):  
N. JEPPSON ◽  
G. XIANGQUN
Keyword(s):  
Ascorbic Acid ◽  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Sea Buckthorn ◽  
Maturation Stages ◽  
The Mean ◽  
Over Time

The contents of kaempherol, quercetin and L-ascorbic acid in sea buckthorn berries were measured at different maturation stages using High Performance Liquid Chromatography (HPLC) methods. The content of ascorbic acid decreased over time with significant differences between sampling dates for the five cultivars studied. The mean decrease was 25% in 19 days, from 1.48 to 1.10 g kg-1. Quercetin decreased whereas kaempherol increased during maturation. Among three studied cultivars, the decrease in quercetin was significant (from 0.028 to 0.014 g kg-1) in 'Otradnaja', where as the increase (from 0.012 to 0.016 g kg-1) in kaempherol was significant in the others two, 'Prozratnaja' and 'Gibrid Pertjik'.;

scholarly journals Sensitive analysis of ethanolamine- and serine-containing phosphoglycerides by high-performance liquid chromatography

1975 ◽  
Vol 145 (3) ◽  
pp. 517-526 ◽  
Author(s):  
F B Jungalwala ◽  
R J Turel ◽  
J E Evans ◽  
R H McCluer
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Limit Of Detection ◽  
High Performance Liquid Chromatographic ◽  
Amino Groups ◽  
Tissue Samples ◽  
Primary Amino ◽  
Liquid Chromatographic Analysis ◽  
Liquid Chromatographic

A highly sensitive method for the separation and quantitative measurement of phospholipids containing primary amino groups, such as phosphatidylethanolamine, phosphatidylserine and lysophosphatidylethanolamine, is described. The method involves a simple and quantitative derivative formation of the phospholipids containing amino groups to their u.v.-absorbing biphenylcarbonyl derivatives. These have molar extinction coefficients of about 23,000 at 268nm. The phospholipid derivatives are then separated and non-destructively determined by high-performance liquid chromatography. The amino phospholipids containing vinyl ether bonds (plasmalogens) can be determined separately from the diacyl- and alkylacyl-amino phospholipids. The lower limit of detection by high-performance liquid-chromatographic analysis of the phospholipid derivatives is about 10-13pmol or 0.3-0.4ng of phospholipid P. The quantitative range of derivative formation and analysis by high-performance liquid chromatography of the phospholipids containing amino groups was shown to be 10-500nmol. The method was shown to be applicable to the analysis of phospholipids containing amino groups in tissue samples.

Analysis of 8-methoxypsoralen by high-performance liquid chromatography

1990 ◽  
Vol 36 (11) ◽  
pp. 1956-1957 ◽  
Author(s):  
C H Ketchum ◽  
C A Robinson ◽  
S T Huang
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Lower Limit ◽  
High Performance ◽  
Limit Of Detection ◽  
Extraction Recovery ◽  
Rapid Procedure ◽  
Standard Curve ◽  
Assay Precision ◽  
Interassay Precision

Abstract We report a simple and rapid procedure for assaying 8-methoxypsoralen (8-MOP) in plasma by high-performance liquid chromatography (HPLC). The standard curve for the assay is linear for 8-MOP from 15 to 500 micrograms/L (y = 0.002x-0.01, r = 0.99) with a lower limit of detection of 1.5 micrograms/L. Intra-assay precision (CV) was 6.0% at the 100 micrograms/L concentration and 10.0% at 50 micrograms/L (n = 30 each). Interassay precision was 6.4% at 100 micrograms/L and 7.0% at 50 micrograms/L (n = 50 each). Extraction recovery of 8-MOP was 98%. Common antiarrhythmics, sedatives, and hypnotics were found not to interfere.

Determination of Chloramphenicol, Thiamphenicol and Florfenicol in Chinese Gelatin Medicines using Dispersive Solid-Phase Extraction Coupled with Ultra High-Performance Liquid Chromatography-Mass Spectrometry.

2020 ◽  
Vol 58 (5) ◽  
pp. 471-476 ◽  
Author(s):  
Jiong Li ◽  
Jinyan Gong ◽  
Haina Yuan ◽  
Gongnian Xiao ◽  
Hongqing Wang ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Matrix Effect ◽  
Analytical Method ◽  
High Performance ◽  
Limit Of Detection ◽  
Solid Phase ◽  
Ammonium Hydroxide ◽  
Drug Residues ◽  
Protein Components

Abstract This study established a rapid and reliable method to determine chloramphenicol (CAP), thiamphenicol (TAP) and florfenicol (FF) residues in Chinese gelatin medicines. CAP, TAP and FF were extracted from medicine samples using 2% (v/v) ammonium hydroxide in acetonitrile. Trypsin was used to eliminate the matrix effect caused by protein components in gelatin medicines, whereas anhydrous sodium sulfate, C18-N and NH2-PSA adsorbents were applied to reduce matrix effect induced by other components. The analytical method of these drugs was optimized on ultra high-performance liquid chromatography-mass spectrometer (UHPLC-MS/MS) through the analysis of their standard linearity and regression. The optimized extraction and analytical method were validated in one Chinese gelatin medicine sample (Colla corii asini, E Jiao) with three fortification levels (2, 5 and 10 μg/kg), and the recoveries of these drug residues ranged of 87.6–102.7%. The limit of detection and quantification of CAP, TAP and FF in the sample were 0.2 and 0.5 μg/kg, 0.4 and 1.5 μg/kg, and 0.5 and 1.5 μg/kg, respectively. A total of 30 Chinese gelatin medicine samples were analyzed using the established method. No drug residues were found in these samples except for one Testudinis Carapacis et Plastri (1.67 μg/kg FF) and one turtle shell glue (2.55 μg/kg FF).

Sign in / Sign up

Export Citation Format

Share Document