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Biomedical subjects

K K Midha

Publications and source records attributed to K K Midha.

At least 163 records · Page 9Linked to original sources
PubMed1985

A comparison of two radioimmunoassays for 7-hydroxychlorpromazine: rabbit polyclonal antibodies vs. mouse monoclonal antibodies.

Two radioimmunoassays (RIAs) were developed for 7-hydroxychlorpromazine (7-OHCPZ), which is a pharmacologically active chlorpromazine (CPZ) metabolite. One of the RIAs used polyclonal antibodies produced in rabbits immunized with a 7-OHCPZ-protein conjugate, which was prepared by coupling 7-hydroxy-N-(2-carboxyethyl)desmethylchlorpromazine to bovine serum albumin by a mixed anhydride method (90% yield). The other RIA was based on mouse monoclonal antibodies produced by hybridomas against the same conjugate. The mouse monoclonal antibodies were considerably more specific than the rabbit polyclonal antibodies. There was little interference with the measurement of 7-OHCPZ by RIA based on mouse monoclonal antibodies even when the samples were spiked with 7-OHCPZ in the presence of five times excess of CPZ and two major metabolites, CPZ sulfoxide and CPZ-N-oxide. By contrast, there was a significant increase in the apparent concentration of 7-OHCPZ when the same samples were assayed by RIA based on polyclonal antibodies. The RIA based on mouse monoclonal antibodies was applied, together with an RIA for CPZ to determine the concentrations of 7-OHCPZ and CPZ in plasma samples from two healthy volunteers after they had received a single 50 mg p.o. dose of CPZ. To our knowledge, this is the first time that plasma 7-OHCPZ concentrations, measured up to 24 hr after a single dose of CPZ, are reported in humans.

Animals↗
PubMed1984

Gas chromatographic quantitation of methoxyphenamine and three of its metabolites in plasma.

Sensitive gas chromatographic procedures for the determination of methoxyphenamine and three of its metabolites in plasma have been developed. The metabolites were measured using an electron-capture detector. This simple procedure is based on the precipitation of protein from a 1-ml plasma sample with 10% trichloroacetic acid, followed by aqueous derivatization with pentafluorobenzoyl chloride at pH 9.2 and a single-step cyclohexane extraction. The lower limit of detection for the N-desmethyl, O-desmethyl and aromatic 5-hydroxy metabolites of methoxyphenamine were 1.6, 3.1 and 2.2 ng ml-1, respectively, with coefficients of variation less than 10%. The poor electron-capture response of fluorinated derivatives of methoxyphenamine necessitated the use of nitrogen-phosphorus detection. Extractive derivatization with pentafluorobenzoyl chloride, without the need for protein precipitation, enabled quantitation of methoxyphenamine down to 3.8 ng ml-1 from a 2-ml aliquot of plasma. In a pilot study involving healthy volunteers who received a single oral dose of methoxyphenamine hydrochloride plasma concentration could be followed in all three subjects for at least 24, 32, 12 and 4 h for methoxyphenamine and the O-desmethyl, 5-hydroxy and N-desmethyl metabolites, respectively.

Adrenergic beta-Agonists↗
PubMed1984

Synthesis of deuterium-labeled fluphenazine.

The propylpiperazine side chain of fluphenazine has been labeled with two, four, and six deuterium atoms by lithium aluminum deuteride reduction of the appropriate ester or imide. The gamma-carbon of the propyl group was labeled with two deuterium atoms by reduction of 10- (2-methoxycarbonylethyl) -2-trifluoromethyl-10H-phenothiazine, while four deuterium atoms were incorporated into the piperazine ring by reduction of 10-[3-(3,5-dioxo-1-piperazinyl)propyl]-2-trifluoromethyl-10H-pheno thiazine. The latter reduction gave the d4-labeled N-deshydroxyethyl metabolite of fluphenazine.

Chemical Phenomena↗
PubMed1984

Radioimmunoassay for psychotropic drugs. III: Synthesis and properties of haptens for trifluoperazine and fluphenazine.

For the development of radioimmunoassay procedures for trifluoperazine and fluphenazine, three haptens, N-(2-carboxyethyl)desmethyltrifluoperazine, N-(4-carboxybutyl)desmethyltrifluoperazine, and 10-[3-(4-carboxyethylpiperazinyl)-3-oxopropyl]-2-trifluoromethyl-+ ++10H- phenothiazine, were synthesized and characterized. Each hapten was coupled to bovine serum albumin, and the number of hapten residues per mole of bovine serum albumin was calculated by UV spectrophotometric methods. Antibodies to each hapten-protein conjugate were developed in rabbits, and titers of the antisera were checked by evaluating their binding characteristics to the tritiated drug.

Animals↗
PubMed1984

Relative bioavailability of a commercial trifluoperazine tablet formulation using a radioimmunoassay technique.

The relative bioavailability of a new conventional tablet formulation (5 mg) of trifluoperazine dihydrochloride was studied in 24 healthy volunteers. Using a sensitive radioimmunoassay technique, plasma trifluoperazine concentrations were measured up until 24 h following ingestion of single 5-mg doses of trifluoperazine. The mean +/- SD for the peak concentration (Cmax), time to Cmax, area under the curve from 0 to 24 h (AUC240), and terminal elimination half-life following the administration of the test formulation were 2.15 +/- 1.07 ng/mL, 4.10 +/- 1.38 h, 21.04 +/- 11.92 ng X h/mL, and 9.5 +/- 7 h, respectively. Following the ingestion of the original trifluoperazine tablet formulation (5 mg) these same parameters were estimated to be 1.92 +/- 0.88 ng/mL, 4.02 +/- 1.10 h, 18.03 +/- 10.11 ng X h/mL, and 9.3 +/- 7 h, respectively. Large intersubject variations in Cmax and AUC240 were observed. The relative bioavailability of the test formulation was calculated to be 106.5 +/- 25.5%.

Adult↗
PubMed1984

Subnanogram quantitation of chlorpheniramine in plasma by a new radioimmunoassay and comparison with a liquid chromatographic method.

A new radioimmunoassay (RIA) procedure for the quantitation of chlorpheniramine in plasma is described. The assay allows the determination of chlorpheniramine levels up to 96 h after oral administration of a single 4-mg tablet to healthy volunteers. This procedure was sensitive to a 156-pg/mL plasma concentration when a 100-microL plasma sample was used. The mean coefficient of variation over the linear range of the assay from 0.156 to 20 ng/mL was 3.79%. The specificity of the assay was investigated, and the antisera showed 7% cross-reactivity with the N,N-didemethyl analogue and 17% cross-reactivity with the N-demethyl analogue. This high degree of specificity was also evident from the findings that the plasma concentrations determined by this newly described RIA procedure in samples of two healthy male volunteers who were administered 4 mg of chlorpheniramine maleate orally gave a strong correlation (r2 = 0.88) with values obtained by an HPLC-UV procedure. The antiserum cross-reacted 100% with brompheniramine and, thus, can be used for its analysis in plasma. The described RIA procedure is precise, simple, and capable of handling a large number of plasma samples with a minimal turnaround time.

Blood Specimen Collection↗
PubMed1984

Radioimmunoassay for the sulfoxide metabolite of trifluoperazine and its application to a kinetic study in humans.

Antibodies were produced in rabbits immunized with 10-[[3-[4-(2-carboxyethyl)-1-piperazinyl]-propyl]]-2 -trifluoromethyl-10H-phenothiazine sulfoxide-bovine serum albumin conjugate. The subsequently developed radioimmunoassay (RIA) procedure enables, for the first time, the quantitation of the sulfoxide metabolite of trifluoperazine in the plasma of humans after administration of therapeutic doses of trifluoperazine, in which 60 pg of the sulfoxide metabolite in 200 microL of plasma can be measured with a CV of less than 3%. Similar results were obtained by this assay with or without a benzene extraction step and also in the presence or absence of a large excess of trifluoperazine and suspected major metabolites of trifluoperazine. This RIA procedure, together with a previously developed RIA for trifluoperazine was used to directly determine plasma concentrations of trifluoperazine and its sulfoxide metabolite after administration of a single, low, oral dose of trifluoperazine to five healthy volunteers. The rapidly appearing, relatively high concentrations of the sulfoxide metabolite are indicative of presystemic sulfoxidation. The mean plasma elimination half-life for the sulfoxide metabolic of trifluoperazine was 5.8 +/- 1.3 h.

Humans↗
PubMed1984

Radioimmunoassay for trimeprazine in human plasma.

Antisera to trimeprazine were raised in New Zealand white rabbits to an immunogen synthesized by covalent linkage of bovine serum albumin to N-(2-carboxyethyl)desmethyltrimeprazine. By use of an antiserum, a radioimmunoassay for trimeprazine was developed that is able to quantitate 0.38 ng/ml-1 in a 200 microliter plasma sample with a coefficient of variation of approximately 12%. The antiserum did not cross-react with the supposedly pharmacologically inactive metabolite trimeprazine sulfoxide; however, the cross-reactivity with the supposedly active metabolite N-desmethyltrimeprazine is significant (49%). The radioimmunoassay was able to measure the drug and/or N-desalkyl metabolites in plasma samples obtained as late as 24 hr following administration of a single oral dose (10mg) of trimeprazine tartrate. Analysis of the same plasma samples by a published high-performance liquid chromatographic procedure gave values much lower than those obtained by the radioimmunoassay, indicating the N-desalkyl metabolites are produced significantly after trimeprazine oral administration.

Antibody Affinity↗
PubMed1984

The effect of sample storage on the stability of chlorpromazine in plasma and whole blood.

The stability of chlorpromazine in heparinized whole blood and plasma was investigated at three different concentrations (5, 20, and 50 ng/ml) after storage at -20 degrees C for 0, 7, 14, 28, 56, and 84 days. A statistical analysis was done to determine whether there was any significant difference between the concentration of chlorpromazine in these body fluids as determined on day one versus each of the later dates of analysis. The results indicate that no significant difference was found over the course of these investigations.

Chlorpromazine↗
PubMed1983

Binding of phenothiazine neuroleptics to plasma proteins.

The binding of chlorpromazine, trifluoperazine, perphenazine, desipramine, propranolol and salicylic acid to human plasma and isolated plasma proteins was studied using equilibrium dialysis. Unlike salicylic acid, an acidic compound only bound to human serum albumin, the basic drugs were bound to all plasma protein fractions studied (albumin, alpha 1-acid glycoprotein, lipoproteins, gamma-globulins) with alpha 1-acid glycoprotein an important binding protein for each of them. The interaction of chloropromazine, perphenazine and trifluoperazine with alpha 1-acid glycoprotein was studied using Scatchard analysis. The primary class of binding sites revealed a low capacity (n = 0.5-1) and a high affinity (K = 10(5)-10(6) M-1) for the phenothiazines. The interaction of chlorpromazine, perphenazine and trifluoperazine with albumin was of the high capacity-low affinity type. In binding studies using plasma obtained from healthy volunteers, alpha 1-acid glycoprotein was found to be a very important binding protein for the basic drug studied with the exception of desipramine. This shows that results derived from binding studies using isolated protein fractions should be interpreted with caution.

Antipsychotic Agents↗
PubMed1983

Subnanogram determination of fluphenazine in human plasma by gas chromatography mass spectrometry.

A new gas chromatographic mass spectrometric procedure for the quantitation of fluphenazine in plasma is described. The method relies on the selected ion monitoring of fluphenazine (m/z 406.1563) and perphenazine (m/z 372.1299), the internal standard, after extraction from plasma with 5% isopropanol in n-pentane. Interferences by plasma constituents such as cholesterol are avoided by including an n-hexane wash. This wash step reduced the recovery of fluphenazine and to a greater extent perphenazine, however, it yielded an organic solution relatively free of any peaks with interfering ions. Prior to gas chromatographic mass spectrometric analysis the silyl derivatives of fluphenazine and perphenazine are prepared using N,O-bis(trimethyl-silyl)acetamide. This procedure allows for the quantitation of as low as 78 pg of fluphenazine per ml of plasma using 2.0 ml plasma aliquots with a coefficient of variation of 4.6%. The high specificity and sensitivity demonstrated by this method allows for the first time the monitoring of plasma concentrations of fluphenazine up to 32 h following a single oral dose of 5 mg of the drug.

Fluphenazine↗
PubMed1983

Synthesis of deuterium-labeled prochlorperazine.

The propylpiperazine side chain of prochlorperazine was labeled with two, four, or six deuterium atoms by lithium aluminum deuteride reduction of the appropriate amide. The isotopic purity of the products after correcting for chlorine isotopes was greater than 95.7%.

Deuterium↗
PubMed1983

Stereospecific radioimmunoassays for l-ephedrine and d-ephedrine in human plasma.

Haptens were prepared by the reaction of d-ephedrine or l-ephedrine with methyl acrylate and subsequent alkaline hydrolysis of the methyl ester groups. The haptens were coupled to bovine serum albumin by a mixed anhydride method, and the resulting drug-protein conjugates were used to immunize rabbits. Antisera raised to these conjugates were highly stereospecific. Neither antiserum cross-reacted with the optical antipode of its substrate nor with racemic pseudoephedrine. Separate radioimmunoassays (RIAs), developed for d-ephedrine and l-ephedrine, were used to measure the concentrations of the enantiomers of ephedrine in the blood of two volunteers dosed with racemic ephedrine. The RIAs were validated by comparing the sum of the concentrations of the enantiomers, determined by RIA, with total ephedrine concentrations determined by a nonstereoselective GLC-ECD method.

Animals↗
PubMed1983

Subnanogram quantitation of chlorpromazine in plasma by high-performance liquid chromatography with electrochemical detection.

A specific and sensitive high-performance liquid chromatographic (HPLC) method for the quantitative determination of subnanogram levels of chlorpromazine in plasma is described. Following extraction of chlorpromazine and the internal standard, prochlorperazine, HPLC analysis is carried out on a cyano column with a mobile phase consisting of 0.1 M ammonium acetate in acetonitrile (10:90 v/v). The use of oxidative thin-layer amperometric detection allowed the quantitation of 0.25 ng of chlorpromazine/ml of plasma with a coefficient of variation of 5.1%. The HPLC method has adequate sensitivity to follow plasma concentration-time profiles up to 24 hr following low single oral doses of chlorpromazine in healthy volunteers.

Chlorpromazine↗
PubMed1983

Kinetics of oral fluphenazine disposition in humans by GC-MS.

The disposition of fluphenazine was investigated in six healthy volunteers following oral administration (5 mg). Using a sensitive and specific GC-MS procedure plasma fluphenazine concentrations were measured up until 32 h after drug administration. Peak plasma concentrations varied widely (range: 0.26-1.06 ng/ml) and were observed at 2.8 +/- 0.5 h following fluphenazine administration. The apparent terminal elimination half-life of fluphenazine was 33.1 +/- 8.1 h. The area under the plasma concentration-time curve differed widely between subjects (range: 7.1-28.6 ng/ml h) suggesting large interindividual differences in the extent of fluphenazine presystemic elimination.

Administration, Oral↗
PubMed1983

Development of radioimmunoassays for trifluoperazine and their application to metabolic studies of the drug.

Antisera to trifluoperazine have been raised in New Zealand white rabbits to several different types of immunogens, where there was variation in the length and nature of the side chain attached to the phenothiazine nucleus, as well as in the number of hapten residues coupled to bovine serum albumin. A radioimmunoassay for trifluoperazine has been developed which is capable of quantitating 0.3125 ng ml-1 in a 200 microliter plasma sample, with cross-reactivities to the sulfoxide, 7-hydroxy, and N-desmethyl metabolites of trifluoperazine of the order of less than 1, 11, and 12%, respectively. Some of the investigated antisera were applied to metabolic studies involving trifluoperazine, where it was demonstrated that N-desmethyltrifluoperazine, rather than 7-hydroxytrifluoperazine, was a major metabolite of trifluoperazine in plasma of a volunteer following administration of a single 5 mg oral dose.

Cross Reactions↗