Impact of creatinine production on the agreement between glomerular filtration rate estimates using cystatin C-derived, and 4- and 6-variable Modification of Diet in Renal Disease (MDRD), and Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equations.

BACKGROUND: It has recently been reported that patient selection has a strong impact on the agreement between glomerular filtration rate (GFR) estimates from serum cystatin C and creatinine. The aim of our study was to evaluate the effect of creatinine production rate (CPR) on this subject. MATERIAL AND METHODS: GFR was estimated from serum cystatin C and from creatinine using the 4- and 6-variable Modification of Diet in Renal Disease (MDRD), and Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equations in 50 healthy subjects, 43 patients with renal failure, 794 kidney and 104 liver transplant recipients, 61 patients with heart failure, 59 patients with biliary obstruction, and 113 critically ill patients. RESULTS: In the 295 patients with impaired CPR (< 900 mg/24 h/1.73 m(2)), discordances of more than 40% between GFR(MDRD4) and GFR(cystatinC) were observed in 38% of cases, between GFR(MDRD6) and GFR(cystatinC) in 22%, and between GFR(CKD-EPI) and GFR(cystatinC) in 27% (in all cases due to GFR overestimation from creatinine). In the 929 patients with maintained CPR (> 900 mg/24 h/1.73 m(2)), greater discordances than 40% between GFR(MDRD4) and GFR(cystatinC) were observed in 8% of cases, between GFR(MDRD6) and GFR(cystatinC) in 9%, and between GFR(CKD-EPI) and GFR(cystatinC) in 7% (in the major part of cases due to GFR overestimation from cystatin C). CONCLUSION: The main source of differences of more than 40% between GFR estimates from serum creatinine and cystatin C is a GFR overestimation in patients with low CPR and GFR underestimation in patients with high CPR by the creatinine-derived equations.

Association between butyrylcholinesterase activity and low-grade systemic inflammation.

BACKGROUND: Pro-inflammatory cytokine production is directly inhibited by acetylcholine (ACh), and a relationship between total circulating ACh hydrolytic capacity and inflammatory reactions has been previously reported. Butyrylcholinesterase (BChE) is the major ACh hydrolyzing enzyme in plasma, and the aim of our study was to evaluate its association with low-grade systemic inflammation. MATERIAL AND METHODS: A total of 4,077 patients clinically managed in the Cardiology, Hypertension, and Digestive Medicine Units were included in our study. Three subclinical chronic inflammatory degrees were established in accordance with the high-sensitivity C-reactive protein (hsCRP) concentrations proposed, for low (< 1 mg/L), average (1-3 mg/L), and high (> 3-10 mg/L) cardiovascular disease risk estimation. RESULTS: In male patients with subclinical chronic inflammation and hsCRP concentrations < 1 mg/L, a significant positive correlation was observed between BChE and hsCRP (p < 0.02); however, for hsCRP concentrations > 3 mg/L, the correlation between these variables in both sexes becomes significantly negative (p < 0.001), as in patients with acute inflammation (hsCRP > 10 mg/L). In all cases significant positive correlations were obtained between the BChE activities and albumin concentrations (p < 0.001). CONCLUSIONS: The liver production of BChE and albumin occurs in a coupled fashion, and these biochemical variables may be considered as negative inflammatory reactants, whose serum levels are inversely associated with the increasing degree of subclinical inflammation.

Determination of everolimus in blood samples from kidney and liver transplant recipients using the sirolimus chemiluminescence magnetic microparticle immunoassay (CMIA) on the Architect-i1000® system.

BACKGROUND: There is significant immunoassay cross-reactivity between everolimus and sirolimus, and their routine determination using a common method may reduce the reagent costs. METHODS: In 122 blood samples from kidney (n = 30) and liver (n = 92) transplant recipients, everolimus concentrations were determined using the Abbott IMx® microparticle enzyme immunoassay (MEIA) as previously described, and the Abbott sirolimus chemiluminescence magnetic microparticle immunoassay (CMIA) on the Architect-i1000® system. RESULTS: A high correlation coefficient (r = 0.981, p < 0.001) and a linear regression MEIA = 0.73CMIA + 0.55, with an acceptable standard error of the estimate (ma68 = 0.32 ng/mL), were obtained, indicating the transferability of the results produced by both immunoassays. CONCLUSIONS: The newly-developed sirolimus CMIA assay on the Architect® platform may be a valid alternative to other immunoassays for the routine therapeutic monitoring of everolimus.

Investigation of possible interference by digoxin-like immunoreactive substances on the Architect iDigoxin CMIA in serum samples from pregnant women, and patients with liver disease, renal insufficiency, critical illness, and kidney and liver transplant.

BACKGROUND: Although the possible interference of digoxin-like immunoreactive substances (DLIS) on the Architect iDigoxin chemiluminiscent microparticle immunoassay (CMIA) has been emphasized by the manufacturer, a specific study about this subject has still not been carried out. METHODS: Apparent serum digoxin concentrations were determined using the Architect iDigoxin CMIA from Abbott Laboratories in digoxin-free pregnant women (n = 50), and patients with liver disease (n = 50), renal insufficiency (n = 50), kidney (n = 25) or liver (n = 25) transplant, and critical illness (n = 50). RESULTS: In all of the patients included in this study, apparent serum digoxin concentrations were lower than the correspondent quantification limit (< 0.30 microg/L). CONCLUSIONS: The Architect iDigoxin CMIA assay would be relatively free from endogenous DLIS positive interferences.

Apparent clearance of valproic acid in elderly epileptic patients: estimation of the confounding effect of albumin concentration.

BACKGROUND: Valproic acid (VPA) apparent clearance (CL) estimated from total serum concentrations is analogous in elderly and non-elderly adult patients. As drug-protein binding decreases in old age, the aim of our study was to evaluate the confounding effect of the serum albumin concentration on the VPA apparent CL in elderly patients. METHODS: In 102 epileptic out-patients treated with VPA in monotherapy, serum total steady-state trough concentrations (Css) were determined. Css concentrations were normalized for a 42 g/L albumin concentration (Css(N)), and the apparent CL and normalized apparent CL(N) were calculated. RESULTS: A poor concordance of 53% was found in the classification of Css and Css(N) levels of VPA as subtherapeutic, therapeutic, or supratherapeutic dose. In the elderly (≥65 years) and non-elderly adult patients, the VPA apparent CL was similar; however, normalized apparent CL(N) was significantly lower in older patients (P < 0.01), with a 40% median decrease. CONCLUSIONS: Total VPA concentrations should be interpreted with caution, mainly in older patients, in which determination of unbound or normalized total drug concentrations may be clinically useful. Normalization of total concentrations permits an estimation of the masking effect of serum albumin concentrations on the VPA apparent CL in elderly patients.

Determination of blood everolimus concentrations in kidney and liver transplant recipients using the sirolimus antibody conjugated magnetic immunoassay (ACMIA).

BACKGROUND: The aim of our study was to evaluate the possible determination of everolimus concentrations using the newly-introduced sirolimus antibody conjugated magnetic immunoassay (ACMIA). METHODS: Everolimus concentrations were determined in 100 blood samples from kidney (n = 47) and liver (n = 53) transplant recipients using the IMx sirolimus microparticle enzyme immunoassay (MEIA) from Abbott as previously described (Clin Biochem 2007;40:132-36) and sirolimus ACMIA from Siemens Healthcare Diagnostics Ltd. RESULTS: The ACMIA everolimus values were significantly higher than those of MEIA (p < 0.001). Analogous slope and intercept values were obtained in the linear regression between the ACMIA and MEIA results when compared to the Seradyn Certican everolimus controls or the blood samples from transplant recipients. Correction of the ACMIA values using the regression equation obtained for the control material (ACMIAcorrected = 0.55 ACMIA + 1.14) led to a satisfactory relationship with the results provided by the MEIA for the patients' samples (MEIA = 1.00 ACMIAcorrected + 0.30, r = 0.905, p < 0.001). CONCLUSIONS: The sirolimus ACMIA on the Dimension platform, which does not require manual pre-treatment of the blood samples, may be an acceptable option for therapeutic everolimus monitoring, significantly reducing technician time in comparison to other widely-used immunoassays.

Macromolecular complexes of cystatin C with circulating liver plasma membrane fragments and determination of serum cystatin C using the particle enhanced nephelometric immunoassay (PENIA) in kidney and liver transplantation and biliary obstruction.

BACKGROUND: In a patient with biliary obstruction, a macromolecular complex of cystatin C with liver plasma membrane fragments, which also contain several membrane-bound enzymes, which may be removed by butanol extraction, has recently been characterised. This could lead to an underestimation of the glomerular filtration rate (GFR) from serum cystatin C concentration. METHODS: Using the particle enhanced nephelometric immunoassay (PENIA), serum cystatin C concentration was determined in 50 healthy controls, 43 patients with renal insufficiency, 68 kidney and 88 liver transplant recipients, and 60 patients with biliary obstruction. Residual cystatin C concentrations and gamma-glutamyltransferase (GGT) activities after butanol extraction were also determined. RESULTS: In the controls and patients with renal insufficiency the residual concentrations of cystatin C after butanol extraction were always > or = 80%. However, in 2 renal and 7 liver transplant recipients and in 11 patients with biliary obstruction this biochemical variable was < 80%. A significant correlation between the residual cystatin C concentrations and residual GGT activities was obtained (r = 0.386, p < 0.001). In the relationship between estimated GFR from serum cystatin C and creatinine concentrations in the liver transplant recipients and patients with biliary obstruction, the 18 cases with residual cystatin C concentrations < 80% were included in the distribution of the total population data with no particular tendency. CONCLUSIONS: These results suggest that in the cases with residual cystatin C concentrations < 80% after butanol extraction, presumably due to the presence of circulating macromolecular cystatin C, the serum levels of cystatin C obtained using the PENIA assay do not lead to a systematic GFR underestimation.

A limited sampling strategy for estimation of the area under the curve (0 to 8 hours) of mycophenolic acid administered three times daily to liver transplant recipients.

OBJECTIVES: Gastrointestinal side-effects caused by mycophenolic acid (MPA) are frequent in liver transplant recipients, and in these cases a switch from two to three daily doses is usually recommended. However, a limited sampling strategy for the estimation of MPA area under the curve from 0 to 8 hours (AUC(0-8h)) has not been made. DESIGN AND METHODS: In 22 liver transplant patients who were administered MPA three times daily, the trapezoidal extrapolated MPA AUC(0-8h) values using a sampling time from 0 to 2 hours were calculated. RESULTS: A tentative therapeutic range for MPA AUC(0-8h) of about 20-40 µg.h/mL is proposed, and in the 13 patients with supratherapeutic values the total leukocyte blood count was significantly lower than in the 9 patients with AUC(0-8h) ≤ 40 µg.h/mL (P < 0.001). Significant negative correlations were found between the total leukocyte blood count and the MPA trough levels (r = -0.458; P < 0.05), AUC(0-8h) (r = -0.479; P < 0.05), and AUC(0-2h) (r = -0.437; P < 0.05). A significant correlation was found between the trapezoidal extrapolated AUC(0-8h) and trapezoidal AUC(0-2h) results (r = 0.850; P < 0.001). CONCLUSIONS: The trapezoidal extrapolated AUC(0-8h), and possibly trapezoidal AUC(0-2h), may be useful for routine therapeutic MPA monitoring in liver transplant recipients in which the dosing frequency is increased from twice to three times a day.

A preliminary investigation on the possible association between diminished copper availability and non-alcoholic fatty liver disease in epileptic patients treated with valproic acid.

BACKGROUND: Patients treated with valproic acid (VPA) present a high incidence of non-alcoholic fatty liver disease (NAFLD) (around 61%). Several recent studies suggest that low copper stores could be associated with NAFLD, and a significant decrease of copper availability in VPA-treated patients has been described. DESIGN AND METHODS: In 101 adult epileptic patients treated with valproic acid in monotherapy (n = 75) and polytherapy (n = 26) the copper availability was evaluated using the specific oxidase activity of ceruloplasmin (activity per unit mass of enzyme protein) and the copper/ceruloplasmin ratio. Copper deficiency was supposed in the cases in which this biochemical variable was smaller than the lower reference limit (333 U/g). RESULTS: The differences between the groups of patients with ceruloplasmin oxidase activity smaller or greater than 333 U/g for the serum levels of aminotransferases, gamma-glutamyltransferase, butyrylcholinesterase, cholesterol, triglycerides, and C-reactive protein, and the APRI and FIB-4 liver fibrosis scores were not statistically significant. Most patients (93%) had low APRI and FIB-4 scores, suggesting absence of significant liver fibrosis. CONCLUSIONS: The results obtained do not confirm the hypothesis of an association between diminished copper availability and NAFLD in patients treated with valproic acid.

Interference of circulating endogenous antibodies on the Dimension® DGNA digoxin immunoassay: elimination with a heterophilic blocking reagent.

Only two cases have been previously described about circulating endogenous antibodies interference on the immunochemical determination of digoxin. In an elderly patient with history of social cat handling was observed a moderate positive interference (about 0.8 ng/mL) on the Dimension® DGNA digoxin immunoassay (capture rabbit antibody), which was eliminated treating the serum samples with a heterophilic blocking reagent. The Architect® immunoassay was not affected.

Effect of valproic acid treatment on copper availability in adult epileptic patients.

OBJECTIVES: In rats a significant increase of the copper biliary excretion is produced by valproic acid administration, however, a conclusive study on the possible appearance of copper deficiency in humans during treatment with this drug has still not been carried out. DESIGN AND METHODS: In 101 adult epileptic patients treated in monotherapy (n=75) and polytherapy (n=26) with valproic acid, and 50 healthy controls, were determined serum copper, immunoreactive ceruloplasmin and its oxidase activity against o-dianisidine, in order to calculate the specific oxidase activity (activity per unit mass of enzyme protein) and copper/ceruloplasmin ratio. RESULTS: Specific oxidase activity of ceruloplasmin and copper/ceruloplasmin ratio were significantly lower in the groups of patients treated with valproic acid than in the controls. Significant correlations were obtained between both biochemical variables (p<0001). In 38% of the patients treated in mono or polytherapy, the specific oxidase activities of ceruloplasmin were smaller than the estimated lower limit of reference. CONCLUSIONS: These results, possible due to a diminished copper content of the circulating ceruloplasmin, suggest that marginal or moderate copper deficiency may have a substantial prevalence among patients treated with valproic acid.

Sirolimus and everolimus clearance in maintenance kidney and liver transplant recipients: diagnostic efficiency of the concentration/dose ratio for the prediction of trough steady-state concentrations.

OBJECTIVES: Therapeutic monitoring of sirolimus and everolimus is necessary in order to minimize adverse side-effects and to ensure effective immunosuppression. A sirolimus-dosing model using the concentration/dose ratio has been previously proposed for kidney transplant patients, and the aim of our study was the evaluation of this single model for the prediction of trough sirolimus and everolimus concentrations. METHODS: Trough steady-state sirolimus concentrations were determined in several blood samples from each of 7 kidney and 9 liver maintenance transplant recipients, and everolimus concentrations from 20 kidney, 17 liver, and 3 kidney/liver maintenance transplant recipients. Predicted sirolimus and everolimus concentrations (Css), corresponding to the doses (D), were calculated using the measured concentrations (Css(0)) and corresponding doses (D(0)) on starting the study: Css = (Css(0))(D)/D(0). RESULTS: The diagnostic efficiency of the predicting model for the correct classification as subtherapeutic, therapeutic, and supratherapeutic values with respect to the experimentally obtained concentrations was 91.3% for sirolimus and 81.4% for everolimus in the kidney transplant patients. In the liver transplant patients the efficiency was 69.2% for sirolimus and 72.6% for everolimus, and in the kidney/liver transplant recipients the efficiency for everolimus was 67.9%. CONCLUSIONS: The model has an acceptable diagnostic efficiency (>80%) for the prediction of sirolimus and everolimus concentrations in kidney transplant recipients, but not in liver transplant recipients. However, considering the wide ranges found for the prediction error of sirolimus and everolimus concentrations, the clinical relevance of this dosing model is weak.

Glomerular filtration rate estimation using the Cockcroft-Gault and modification of diet in renal disease formulas for digoxin dose adjustment in patients with heart failure.

OBJECTIVE: The aim of this study was to compare the estimated glomerular filtration rate (GFR) using the Cockcroft-Gault and the 4-, 5-, and 6-variable Modification of Diet in Renal Disease (MDRD) formulas for digoxin dose adjustment. METHODS: Steady-state serum digoxin concentrations were determined in 100 patients with heart failure and normal to severely impaired renal function. Total clearance (CL) and predicted average concentrations of digoxin were calculated using general pharmacokinetic principles. RESULTS: The mean+/-SEM (median) estimated GFR values were 48.9+/-2.8 (46.5) mL/min/1.73 m(2) using the Cockcroft-Gault formula, 61.4+/-3.6 (56.4) mL/min/1.73 m(2) using the MDRD4 formula, 56.8+/-3.3 (52.1) mL/min/1.73 m(2) using the MDRD5 formula, and 53.3+/-3.0 (48.7) mL/min/1.73 m(2) using the MDRD6 formula, with high correlation coefficients between the estimates (r > or = 0.928, P < 0.001). Significant correlations were found between the digoxin total CL and estimated GFR by the Cockcroft-Gault (r = 0.649, P < 0.001), MDRD4 (r = 0.634, P <0.001), MDRD5 (r = 0.635, P < 0.001), and MDRD6 (r = 0.652, P < 0.001) formulas. A significant negative correlation of the digoxin total CL/GFR ratio with estimated GFR was obtained (r = -0.356, P < 0.001), with a high variability for this ratio for GFR lower than 60 mL/min. Analogous correlation coefficients were found between the obtained and predicted digoxin concentrations calculated using the estimated GFR by the Cockcroft-Gault (r = 0.628, P < 0.001), MDRD4 (r = 0.642, P < 0.001), MDRD5 (r = 0.650, P < 0.001), and MDRD6 (r = 0.665, P < 0.001) formulas, with a wide dispersion between the values in all cases. CONCLUSION: For GFR lower than 60 mL/min, the high interindividual variation of the digoxin total CL found among patients with similar renal function is an important limiting factor in the prediction of digoxin dosage regimens.

Falsely increased blood tacrolimus concentrations using the ACMIA assay due to circulating endogenous antibodies in a liver transplant recipient: a tentative approach to obtaining reliable results.

The presence of endogenous antibodies in the serum of some patients has long been known to be a potential source of interference in immunoassays. We report falsely increased whole blood tacrolimus concentrations using the antibody conjugated magnetic immunoassay from Siemens HealthCare Diagnostics in a liver transplant recipient due to the presence of circulating endogenous antibodies (possibly heterophilic antibodies). Estimation of whole blood tacrolimus concentrations from the washed erythrocytes concentrations is proposed as a tentative approach for obtaining reliable results using the antibody conjugated magnetic immunoassay assay in these cases, leading to analogous blood tacrolimus concentrations to those produced by the microparticle enzyme immunoassay from Abbott Laboratories.

Determination of blood sirolimus concentrations in liver and kidney transplant recipients using the Innofluor fluorescence polarization immunoassay: comparison with the microparticle enzyme immunoassay and high-performance liquid chromatography-ultraviolet method.

BACKGROUND: Although high-performance liquid chromatography (HPLC) is the method of choice for blood sirolimus determination, the microparticle enzyme immunoassay (MEIA) run on the IMx analyser is widely used in therapeutic monitoring of this immunosuppressant agent. The aim of our study was to evaluate the possible determination of sirolimus using the fluorescence polarization immunoassay (FPIA) commercialized for everolimus quantification. METHODS: Sirolimus concentrations were determined in whole-blood samples from liver and kidney transplant recipients using the Innofluor Certican FPIA (Seradyn Inc.) run on a TDx analyser (Abbott Laboratories), Sirolimus MEIA run on an IMx analyser (Abbott Laboratories), and HPLC (UV detection) methods. RESULTS: The Innofluor FPIA has a similar cross-reactivity with everolimus and sirolimus, and the within- and between-run coefficients of variation obtained for sirolimus determination were 2.7%-13.3%. In analysing different blood samples from liver and kidney transplant patients the linear regressions obtained were: FPIA = 1.12 HPLC + 0.43 (n=104, r=0.874), MEIA = 1.14 HPLC (n=146, r=0.892), and FPIA = 1.00 MEIA + 0.29 (n=106, r=0.941). Better correlation coefficients were obtained between the methods in the liver transplant samples (r>or=0.900) than in the kidney transplant samples (r>or=0.849). No significant effect was found for sirolimus clearance or the blood hematocrit on the relationship between the results produced by both immunoassays and HPLC. CONCLUSION: The Innofluor FPIA is a valid alternative with an analogous performance to the MEIA for the therapeutic monitoring of sirolimus.

Comparison of blood tacrolimus concentrations in liver and kidney transplant recipients using ACMIA and MEIA immunoassays.

BACKGROUND: Published data on the performance of the new Dade Behring antibody conjugated magnetic immunoassay (ACMIA) for tacrolimus determination are scarce. The aim of this study was to compare the results obtained using the ACMIA and Abbott microparticle enzyme immunoassay (MEIA), which is the most widely used method for therapeutic tacrolimus monitoring. METHODS: Trough tacrolimus concentrations were determined in 305 blood samples from kidney (n=138) and liver (n=167) transplant recipients using the ACMIA and MEIA immunoassays. The MEIA results were corrected for hematocrit values lesser than 30% and higher than 40% (Hermida et al. Clin Lab 2005; 51: 43-45). RESULTS: The obtained ACMIA within- and between-run variation coefficients (<10.8%) were acceptable. In the comparison between ACMIA and MEIA results in the blood samples studied, the regression equation was: ACMIA=1.02MEIA+0.29 (r=0.912, p<0.001), with an acceptable difference between the means (8.13+/-0.53 ng/mL vs. 7.62+/-0.50 ng/mL). However, in accordance with the well-established interference of the hematocrit on the MEIA results, a highly significant negative correlation between the MEIA/ACMIA ratio and the hematocrit values was obtained (r=-0.585, p<0.001). When the MEIA results were corrected according to the hematocrit (MEIAHtC), the regression with ACMIA levels was: ACMIA=1.08MEIAHtC-0.09 (r=0.926, p<0.001). This equation was analogous to that obtained between ACMIA and MEIA tacrolimus concentrations in the 164 blood samples with hematocrit of 30-40%. CONCLUSIONS: ACMIA is an acceptable option for therapeutic tacrolimus monitoring, with an important decrease in technician time in relation to the widely used MEIA.

LDL cholesterol estimation using the Anandaraja's and Friedewald's formulas in schizophrenic patients treated with antipsychotic drugs.

OBJECTIVES: The use of new antipsychotic drugs is associated with an increased risk of diabetes and metabolic syndrome, and the routine monitoring of blood lipids during treatment has been recommended. Recently, a new formula for the estimation of low-density lipoprotein (LDL) cholesterol from total cholesterol and triglycerides has been proposed by Anandaraja et al. (Int J Cardiol 2005; 102: 117), and the aim of our study was its evaluation in schizophrenic patients treated with antipsychotic drugs. MATERIALS AND METHODS: In 487 serum samples from schizophrenic patients treated with clozapine in polytherapy, the concentrations of LDL cholesterol were determined by agar gel electrophoresis and the formula of Friedewald et al. (Clin Chem 1972; 18: 499), and compared with the results of the Anandaraja's formula. RESULTS: A higher correlation and lower error of the estimate of the electrophoresis results was found with those of Friedewald (r=0.940, ma68=0.17 mmol/L) than those of Anandaraja (r=0.811, ma68=0.31 mmol/L). Similar results were obtained on making a dichotomy of the patients with and without metabolic syndrome lipid profile. A highly significant correlation was found between the high-density lipoprotein (HDL) cholesterol levels and the Anandaraja/Electrophoresis (r=0.817, p<0.001) and Anandaraja/Friedewald (r=0.977, p<0.001) ratios. CONCLUSIONS: According to our data, Anandaraja's formula tends towards an overestimation or underestimation of LDL cholesterol levels, depending on whether the HDL cholesterol levels are high or low, which may be clinically significant. These results do not support the proposed better accuracy of the Anandaraja's than the Friedewald's formula.

Relative proportions of serum carbamazepine and its pharmacologically active 10,11-epoxy derivative: effect of polytherapy and renal insufficiency.

BACKGROUND: The proposed action mechanism and pharmacological activity of carbamazepine (CBZ) and its major metabolite, carbamazepine-10,11-epoxide (CBZE), are the same. The aim of our study was the investigation of the effect of concomitant antiepileptic treatment and renal insufficiency on the relative proportions of serum CBZ and CBZE. METHODS: Serum trough steady-state CBZ and CBZE concentrations were determined by high-performance liquid chromatography (HPLC) in 140 epileptic patients treated with CBZ in monotherapy (n=100) and polytherapy with phenytoin, phenobarbital and valproate (n=40). The levels of CBZ were also determined using the Dade Behring enzyme multiplied immunoassay technique (EMIT). The glomerular filtration rate (GFR) was estimated from serum cystatin C using the Dade Behring nephelometric immunoassay. RESULTS: The CBZE/CBZ and CBZ+CBZE/CBZEMIT ratios were significantly increased in 7 cases (3 in monotherapy and 4 in polytherapy) with GFR<60 mL/min/1.73m2 in relation to the patients treated in monotherapy or polytherapy having normal or mildly decreased renal function (p<0.001). CONCLUSIONS: In patients with moderate to severe renal insufficiency the relative proportion of CBZE with respect to the parent drug is significantly increased. In these cases, the CBZ concentrations obtained using the EMIT, or other immunoassays having low CBZE cross-reactivity, may have an inadequate diagnostic efficiency.

Evaluation of three dosing models for the prediction of steady-state trough clozapine concentrations.

OBJECTIVE: Therapeutic drug monitoring of clozapine may be useful for the clinical management of schizophrenic patients treated with this atypical antipsychotic drug. The aim of our study was the evaluation of three models for the prediction of steady-state trough clozapine concentration. PATIENTS AND METHODS: The trough serum concentrations of clozapine and norclozapine were determined by high-performance liquid chromatography in 296 samples from a group of 21 schizophrenic patients selected for their good therapeutic compliance. Also, the predicted clozapine concentrations were estimated by applying the models of Oyewumi et al. (Ther Drug Monit 1995; 17: 137), Perry et al. (Biol Psychiatry 1998; 44: 733) and Rostami-Hodjegan et al. (J Clin Psychopharmacol 2004; 24: 70). RESULTS: The efficiency for the accurate estimation of clozapine concentrations as subtherapeutic (<240 ng/mL), therapeutic (240-750 ng/mL) or supratherapeutic (>750 ng/mL), using the models of Oyewumi et al., Perry et al., and Rostami-Hodjegan et al., was 82%, 71% and 77% respectively. CONCLUSIONS: The predictive model of Oyewumi et al., which shows an easy calculation way and the greater diagnostic efficiency, may be of clinical value for the prediction of clozapine concentration or the dose required to achieve a specified concentration.

Clozapine and norclozapine concentrations in serum and plasma samples from schizophrenic patients.

At present, the determination of steady-state trough serum/plasma concentrations of clozapine is considered a useful tool for the clinical management of schizophrenic patients treated with this drug. In a previously published study, it was indicated that only plasma should be used to avoid a significant underestimation of clozapine and norclozapine concentrations; however, a formal evaluation of this topic has still not been made, and a consensus on the use of plasma or serum for therapeutic clozapine monitoring may be desirable. Paired samples of serum and plasma (K3EDTA solution contained in Vacutainer tubes) were obtained from 40 schizophrenic patients, and clozapine and norclozapine concentrations were determined by high-performance liquid chromatography. For the parent drug and its metabolite, serum concentrations were higher than in plasma (approximately 7%), although the correction of plasma concentrations in function of hematocrit values reduced this difference to 3%. High correlation coefficients were found between the serum and uncorrected or corrected plasma clozapine concentrations (r = 0.996, P < 0.001), with clinically acceptable differences between the means and standard error of the estimate and consequently with transferability of the results. The clozapine and norclozapine concentrations in five lithium heparin-containing plasma samples (371.9 +/- 226.7 ng/mL and 217.9 +/- 113.1 ng/mL) were analogous to the corresponding hematocrit-corrected EDTA-containing plasma values (374.4 +/- 225.4 ng/mL and 223.5 +/- 115.2 ng/mL), with correlation coefficients of r > or = 0.998 (P < 0.001). Serum or plasma samples may be used for the therapeutic monitoring of clozapine, and no practical advantages have been found with regard to the stability of the drug or imprecision obtained by using either type of biological matrix.