ABSTRACT
Here, we present the proof-of-concept of a lateral flow assay (LFA) that is capable of detecting small-molecule targets in a noncompetitive manner by deploying a sandwich-type format based on the aptamer kissing complex (AKC) strategy. A fluorescently labeled hairpin aptamer served as the signaling agent, while a specific RNA hairpin grafted onto the strip served as the capture element. The hairpin aptamer switched from an unfolded to a folded form in the presence of the target, resulting in kissing interactions between the loops of the reporter and the capture agents. This design triggered a target-dependent fluorescent signal at the test line. The AKC-based LFA was developed for the detection of adenosine, achieving a detection limit in the micromolar range. The assay revealed the presence of the same analyte in urine. The method also proved effective with another small molecule (theophylline). We believe that the AKC-based LFA approach could overcome many of the shortcomings associated with conventional signal-off methods and competitive processes.
Subject(s)
Adenosine , Aptamers, Nucleotide , Biosensing Techniques , Aptamers, Nucleotide/chemistry , Adenosine/analysis , Adenosine/urine , Biosensing Techniques/methods , Humans , Theophylline/analysis , Theophylline/urine , Limit of Detection , Fluorescent Dyes/chemistryABSTRACT
OBJECTIVE: To investigate the relations between caffeine-derived metabolites (methylxanthines) and plasma lipids by use of population-based data from 2 European countries. METHODS: Families were randomly selected from the general population of northern Belgium (FLEMENGHO), from August 12, 1985, until November 22, 1990, and 3 Swiss cities (SKIPOGH), from November 25, 2009, through April 4, 2013. We measured plasma concentrations (FLEMENGHO, SKIPOGH) and 24-hour urinary excretions (SKIPOGH) of 4 methylxanthines-caffeine, paraxanthine, theobromine, and theophylline-using ultra-high-performance liquid chromatography-tandem mass spectrometry. We used enzymatic methods to estimate total cholesterol, high-density lipoprotein cholesterol, and triglyceride levels and the Friedewald equation for low-density lipoprotein cholesterol levels in plasma. We applied sex-specific mixed models to investigate associations between methylxanthines and plasma lipids, adjusting for major confounders. RESULTS: In both FLEMENGHO (N=1987; 1055 [53%] female participants) and SKIPOGH (N=990; 523 [53%] female participants), total cholesterol, low-density lipoprotein cholesterol, and triglyceride levels increased across quartiles of plasma caffeine, paraxanthine, and theophylline (total cholesterol levels by caffeine quartiles in FLEMENGHO, male participants: 5.01±0.06 mmol/L, 5.05±0.06 mmol/L, 5.27±0.06 mmol/L, 5.62±0.06 mmol/L; female participants: 5.24±0.06 mmol/L, 5.15±0.05 mmol/L, 5.25±0.05 mmol/L, 5.42±0.05 mmol/L). Similar results were observed using urinary methylxanthines in SKIPOGH (total cholesterol levels by caffeine quartiles, male participants: 4.54±0.08 mmol/L, 4.94±0.08 mmol/L, 4.87±0.08 mmol/L, 5.27±0.09 mmol/L; female participants: 5.12±0.07 mmol/L, 5.21±0.07 mmol/L, 5.28±0.05 mmol/L, 5.28±0.07 mmol/L). Furthermore, urinary caffeine and theophylline were positively associated with high-density lipoprotein cholesterol in SKIPOGH male participants. CONCLUSION: Plasma and urinary caffeine, paraxanthine, and theophylline were positively associated with plasma lipids, whereas the associations involving theobromine were less clear. We postulate that the positive association between caffeine intake and plasma lipids may be related to the sympathomimetic function of methylxanthines, mitigating the overall health-beneficial effect of caffeine intake.
Subject(s)
Caffeine/adverse effects , Lipids/blood , Adult , Belgium , Caffeine/blood , Caffeine/metabolism , Caffeine/urine , Cholesterol/blood , Cholesterol, HDL/blood , Chromatography, High Pressure Liquid , Female , Humans , Male , Middle Aged , Switzerland , Tandem Mass Spectrometry , Theobromine/adverse effects , Theobromine/blood , Theobromine/urine , Theophylline/adverse effects , Theophylline/blood , Theophylline/urine , Triglycerides/blood , Xanthines/adverse effects , Xanthines/blood , Xanthines/urineABSTRACT
In-tube solid phase microextraction (IT-SPME) coupled on-line to capillary liquid chromatography with diode array detection provides a simple and fast analytical methodology for the simultaneous quantitation of caffeine and its three primary metabolites (theobromine, paraxanthine and theophylline) in micro samples of serum, saliva and urine matrices. The sample amount required for one analysis was only 2.5⯵L of saliva, 6.25⯵L of serum or 40⯵L of urine, a requirement for its implementation in a hospital laboratory for preterm newborns, where sample availability is a major problem. In standard conditions, 25⯵L of diluted saliva or serum (or 100⯵L of urine) were processed by IT-SPME in 30â¯cm of commercially available capillary GC column coated with ZB-FFAP (100% nitroterephthalic modified polyethylene glycol). The retained compounds were desorbed from the IT-SPME capillary by the mobile phase (a gradient mixture of water and methanol) and the separation was carried out in a C18 column (150â¯mmâ¯×â¯0.5â¯mm i.d., 5⯵m particle size). Analytes eluted before 14â¯min, at a flow rate of 15⯵Lâ¯min-1, and were detected by absorbance at 275â¯nm. The calibration graphs presented good linearity (R2â¯>â¯0.99), without the presence of matrix effect, and recoveries between 84 and 112% were obtained. Limits of detection (S/Nâ¯=â¯3) were 0.1⯵g·mL-1 in serum and 0.5⯵g·mL-1 in saliva and urine samples, for all compounds, and the intra- and inter-day variation coefficients (nâ¯=â¯3) were between 3 and 17%. Analytical figures of merit were similar to those proposed by other methodologies, but using lower sample volume and a faster and simpler sample treatment and analysis. Paired samples of serum and saliva from preterm newborns treated with caffeine at the pediatric intensive care unit were analyzed by the method, with statistically equivalent results for caffeine concentrations.
Subject(s)
Caffeine/chemistry , Caffeine/metabolism , Caffeine/urine , Calibration , Chromatography, Liquid/methods , Humans , Saliva/chemistry , Solid Phase Microextraction/methods , Theobromine/chemistry , Theobromine/metabolism , Theobromine/urine , Theophylline/chemistry , Theophylline/metabolism , Theophylline/urine , Urine/chemistryABSTRACT
Lanthanum oxide nanomaterials were decorated with carbon black (CB) and grafted with a poly(acrylic acid) nanogel to obtain a composite material (CB-g-PAA/La2O3) for simultaneous determination of acetaminophen (AMP), naproxen (NPX), and theophylline (TPH). The nanogel was synthesized by in-situ free radical polymerization. The composite was dropped onto a glassy carbon electrode (GCE), and the modified GCE displays robust electrocatalytic activity towards AMP, NPX, and TPH, with voltammetric signals that are enhanced compared to a bare GCE. Features of merit for AMP, NPX, and TPH, respectively, include (a) peak potentials of 0.42, 0.85 and 0.12 V (vs. Ag/AgCl), (b) linear ranges from 0.05-887, 0.05-884, and 0.02-888 µM, and (c) detection limits of 20, 35, and 15 nM. The practical applicability of the CB-g-PAA/La2O3/GCE was illustrated by analyzing serum and urine samples. Graphical abstract Schematic presentation of simultaneous electrochemical sensing of acetaminophen (AMP), naproxen (NPX), and theophylline (TPH) in real sample analysis using poly(acrylic acid) nanogel covalently grafted onto a carbon black/La2O3 composite (CB-g-PAA/La2O3/GCE).
Subject(s)
Acetaminophen/analysis , Acrylic Resins/chemistry , Lanthanum/chemistry , Nanogels/chemistry , Naproxen/analysis , Oxides/chemistry , Soot/chemistry , Theophylline/analysis , Acetaminophen/blood , Acetaminophen/urine , Electrochemistry , Electrodes , Humans , Models, Molecular , Molecular Conformation , Naproxen/blood , Naproxen/urine , Polymerization , Theophylline/blood , Theophylline/urineABSTRACT
AIMS: The purpose of this study was to explore clinical markers reflecting developmental changes in drug clearance by preterm infants. METHODS: Preterm infants administered aminophylline or theophylline to treat apnoea of prematurity were enrolled in this study. Trough and one of 2 h, 4 h or 6 h post-dose blood samples and urine samples were collected during steady state, to determine the concentrations of theophylline and its targeted metabolites. CYP1A2*1C and CYP1A2*1F genotypes were analyzed. Total, renal and nonrenal clearances of theophylline were calculated, and cytochrome P450 1A2 (CYP1A2) activity was obtained from the ratio of 1-methyluric acid and 3-methylxanthine to theophylline in urine. Multiple linear regression analysis was performed to evaluate the relationships between theophylline clearance and the clinical characteristics of preterm infants. RESULTS: A total of 152 samples from 104 preterm infants were analyzed. A strong association between the serum trough and urine theophylline concentrations was found (P < 0.001). Total, renal and nonrenal clearances of theophylline were 0.50 ± 0.29 ml kg-1 min-1 , 0.16 ± 0.06 ml kg-1 min-1 and 0.34 ± 0.28 ml kg-1 min-1 , respectively. CYP1A2 activity correlated positively with the postnatal age and postmenstrual age. However, CYP1A2 genotype was not associated with CYP1A2 activity, which was significantly associated with nonrenal clearance. CYP1A2 activity, postnatal age , weight and 24-h urine output were significantly associated with total theophylline clearance. CONCLUSIONS: CYP1A2 activity can be monitored using noninvasive random urine samples, and it can be used to assess developmental changes in theophylline clearance by preterm infants.
Subject(s)
Bronchodilator Agents/blood , Bronchodilator Agents/urine , Cytochrome P-450 CYP1A2/metabolism , Theophylline/blood , Theophylline/urine , Aging/metabolism , Apnea/drug therapy , Bronchodilator Agents/therapeutic use , Cytochrome P-450 CYP1A2/genetics , Female , Genotype , Gestational Age , Humans , Infant , Infant, Newborn , Infant, Premature , Male , Theophylline/therapeutic use , Uric Acid/analogs & derivatives , Uric Acid/urine , Xanthines/urineABSTRACT
Theophylline is an inexpensive drug employed in asthma and chronic obstructive pulmonary disorder medications and is toxic at higher concentration. The development of a molecularly imprinted polymer based theophylline electrochemical sensor on glassy carbon electrode by the electropolymerization of 4-amino-5-hydroxy-2,7-naphthalenedisulfonic acid is being discussed in this work. The MIP modification enhances the theophylline recognition ability and the electron transfer kinetics of the bare electrode. The parameters, controlling the performance of the imprinted polymer based sensor, like number of electropolymerization cycles, composition of the pre-polymerization mixture, pH and immersion time were investigated and optimized. The interaction energy and the most stable conformation of the template-monomer complex in the pre-polymerization mixture were determined computationally using ab initio calculations based on density functional theory. The amperometric measurements showed that the developed sensor has a method detection limit of 0.32µM for the dynamic range of 0.4 to 17µM, at optimized conditions. The transducer possesses appreciable selectivity in the presence of structurally similar interferents such as theobromine, caffeine and doxofylline. The developed sensor showed remarkable stability and reproducibility and was also successfully employed in theophylline detection from commercially available tablets.
Subject(s)
Electrochemical Techniques , Molecular Imprinting , Polymers/chemistry , Theophylline/analysis , Caffeine/chemistry , Carbon/chemistry , Dielectric Spectroscopy , Electrodes , Humans , Hydrogen-Ion Concentration , Limit of Detection , Microscopy, Atomic Force , Theobromine/chemistry , Theophylline/analogs & derivatives , Theophylline/blood , Theophylline/chemistry , Theophylline/urine , Tin Compounds/chemistryABSTRACT
In the article by Guessous et al (Guessous I, Pruijm M, Ponte B, Ackermann D, Ehret G, Ansermot N, Vuistiner P, Staessen J, Gu Y, Paccaud F, Mohaupt M, Vogt B, Pechère-Bertschi A, Martin PY, Burnier M, Eap CB, Bochud M. Associations of ambulatory blood pressure with urinary caffeine and caffeine metabolite excretions. Hypertension. 2015;65:691696. doi: 10.1161/HYPERTENSIONAHA.114.04512), which published online ahead of print December 8, 2014, and appeared in the March 2015 issue of the journal, a correction was needed.One of the author surnames was misspelled. Antoinette Pechère-Berstchi has been corrected to read Antoinette Pechère-Bertschi.The authors apologize for this error.
Subject(s)
Blood Pressure Monitoring, Ambulatory , Blood Pressure/physiology , Caffeine/urine , Hypertension/physiopathology , Theobromine/urine , Theophylline/urine , Adult , Biomarkers/urine , Circadian Rhythm/physiology , Cross-Sectional Studies , Female , Humans , Hypertension/urine , Male , Middle Aged , PrevalenceABSTRACT
BACKGROUND: CYP1A2 and CYP2A6 are polymorphic enzymes that metabolise several compounds of clinical importance. This study investigated the prevalent phenotypes of these enzymes and the influence of age and sex on enzyme activity in a Nigerian population. METHODS: Caffeine (110 mg) was administered to each of 129 healthy, unrelated subjects (85 males and 44 females) who were non-smokers. Urine voided within 7 h after caffeine administration was collected for a high performance liquid chromatographic assay of caffeine (137X), 1,7-dimethyluric acid (17U) and 1,7-dimethylxanthine (17X). CYP1A2 activity was measured as a ratio of (17U+17X) to 137X, while 17U/17X served as marker for CYP2A6. Transformed data were analysed and the influences of age and sex on activity were also determined. RESULTS: Distribution of CYP1A2 activity in the population was bimodal with a mean±SD of 0.82±0.41, while that of CYP2A6 was trimodal with a mean±SD activity of 0.27±0.42 of the log-transformed urinary molar ratio of metabolites. The influences of age and sex on enzyme activity for both CYP1A2 and CYP2A6 were not significant (p>0.05). CONCLUSIONS: The study established the prevalence of polymorphism in phenotypes of CYP1A2 and CYP2A6 activity in the Nigerian population, but no influence of age and sex on enzyme activity was observed in this population.
Subject(s)
Caffeine/pharmacokinetics , Cytochrome P-450 CYP1A2/metabolism , Cytochrome P-450 CYP2A6/metabolism , Adolescent , Adult , Age Factors , Algorithms , Biomarkers/urine , Biotransformation , Caffeine/administration & dosage , Caffeine/urine , Coffee , Female , Food, Preserved , Humans , Male , Nigeria , Sex Characteristics , Theophylline/urine , Uric Acid/analogs & derivatives , Uric Acid/urine , Young AdultABSTRACT
BACKGROUND: Caffeine is a widely consumed psychoactive stimulant and is of epidemiologic interest. Major sources of caffeine are challenging to standardize, and the use of biomarkers is proposed as an alternative means of assessing intake. OBJECTIVE: We described urine caffeine and caffeine metabolite concentrations (n = 2466) and excretion rates (n = 2261) in the US population ≥6 y by age, sex, race-ethnicity, and caffeine intake (from foods, beverages, and dietary supplements). METHODS: We measured caffeine and 14 of its metabolites in spot urine samples from the cross-sectional NHANES 2009-2010 by use of LC-tandem mass spectrometry. RESULTS: Caffeine and its metabolites were detectable in the urine of most persons, generally at concentrations ≥1 µmol/L. Median concentrations (95% CI) ranged from 0.560 (0.497, 0.620) µmol/L to 58.6 (48.6, 67.2) µmol/L; median excretion rates from 0.423 (0.385, 0.468) nmol/min to 46.0 (40.7, 50.2) nmol/min. Urine concentrations and excretion rates for 9 analytes (caffeine, theophylline, paraxanthine, 1-methylxanthine, 1-methyluric acid, 1,3-dimethyluric acid, 1,7-dimethyluric acid, 1,3,7-trimethyluric acid, and 5-acetylamino-6-amino-3-methyluracil) had moderate correlations with caffeine intake (Spearman ρ = 0.55-0.68, P < 0.0001); the remaining analytes had low correlations (ρ = 0.15-0.33, P < 0.0001). We observed larger differences in geometric mean concentrations and excretion rates between the highest vs. lowest quartiles of caffeine intake for these 9 compounds than the rest. Consistent with dietary caffeine intake, we observed that urine concentrations and excretion rates for most compounds were significantly (P < 0.05) higher in men than women, non-Hispanic whites than Hispanics and non-Hispanic blacks, and highest in persons aged 40-59 y. CONCLUSION: Excretion of caffeine and its metabolites in urine is common in the US population. According to the observed associations between spot urine concentrations or excretion rates with caffeine intake, several of these compounds show promise as potential biomarkers of caffeine intake.
Subject(s)
Caffeine/administration & dosage , Caffeine/urine , Adolescent , Adult , Black or African American , Biomarkers/urine , Child , Chromatography, Liquid , Cross-Sectional Studies , Female , Hispanic or Latino , Humans , Male , Middle Aged , Nutrition Surveys , Tandem Mass Spectrometry , Theophylline/urine , Uracil/analogs & derivatives , Uracil/urine , Uric Acid/analogs & derivatives , Uric Acid/urine , White People , Xanthines/urine , Young AdultSubject(s)
Blood Pressure Monitoring, Ambulatory , Blood Pressure/physiology , Caffeine/urine , Theobromine/urine , Theophylline/urine , Female , Humans , MaleABSTRACT
Intake of caffeinated beverages might be associated with reduced cardiovascular mortality possibly via the lowering of blood pressure. We estimated the association of ambulatory blood pressure with urinary caffeine and caffeine metabolites in a population-based sample. Families were randomly selected from the general population of Swiss cities. Ambulatory blood pressure monitoring was conducted using validated devices. Urinary caffeine, paraxanthine, theophylline, and theobromine excretions were measured in 24 hours urine using ultrahigh performance liquid chromatography tandem mass spectrometry. We used mixed models to explore the associations of urinary excretions with blood pressure although adjusting for major confounders. The 836 participants (48.9% men) included in this analysis had mean age of 47.8 and mean 24-hour systolic and diastolic blood pressure of 120.1 and 78.0 mm Hg. For each doubling of caffeine excretion, 24-hour and night-time systolic blood pressure decreased by 0.642 and 1.107 mm Hg (both P values <0.040). Similar inverse associations were observed for paraxanthine and theophylline. Adjusted night-time systolic blood pressure in the first (lowest), second, third, and fourth (highest) quartile of paraxanthine urinary excretions were 110.3, 107.3, 107.3, and 105.1 mm Hg, respectively (P trend <0.05). No associations of urinary excretions with diastolic blood pressure were generally found, and theobromine excretion was not associated with blood pressure. Anti-hypertensive therapy, diabetes mellitus, and alcohol consumption modify the association of caffeine urinary excretion with systolic blood pressure. Ambulatory systolic blood pressure was inversely associated with urinary excretions of caffeine and other caffeine metabolites. Our results are compatible with a potential protective effect of caffeine on blood pressure.
Subject(s)
Blood Pressure Monitoring, Ambulatory , Blood Pressure/physiology , Caffeine/urine , Theobromine/urine , Theophylline/urine , Adult , Biomarkers/urine , Blood Pressure/drug effects , Caffeine/pharmacology , Circadian Rhythm/physiology , Cross-Sectional Studies , Female , Humans , Hypertension/epidemiology , Hypertension/prevention & control , Hypertension/urine , Male , Middle Aged , PrevalenceABSTRACT
To study the metabolite excretion of theophylline, a rapid and specific method by liquid chromatography with heated electrospray ionization tandem mass spectrometry (LC-HESI/MS/MS) method for simultaneous determination of theophylline, 1, 3-dimethyluric acid (1,3-DMU), 3-methylxanthine (3-MX) and 1-methyluric acid (1-MU) in human urine was developed using theophylline-d6 and 5-fluorouracil as internal standards. Selected reaction monitoring (SRM) with heated electrospray ionization (HESI) was used in the negative mode for mass spectrometric detection. After diluted with methanol and centrifuged, the analytes and ISs were separated on a XDB-Phenyl (150 mm x 4.6 mm, 5 microm) column with a mixture of water-methanol-formic acid (30 : 70 : 0.15) as mobile phase at a flow rate of 0.6 mL x min(-1). The linear calibration curves for theophylline, 1, 3-DMU, 3-MX and 1-MU were obtained in the concentration range of 1.0-250 microg x mL(-1), separately. The method herein described is effective and convenient, and can be used for determination of theophylline and its three metabolites. The results showed that urinary excretion ratio of theophylline, 1,3-DMU, 3-MX and 1-MU is approximately 1 : 3 : 1 : 2 in Chinese subjects, which is similar to the reported excretion pattern in Caucasian.
Subject(s)
Theophylline/urine , Administration, Oral , Asian People , Calibration , Chromatography, Liquid , Delayed-Action Preparations/metabolism , Healthy Volunteers , Humans , Spectrometry, Mass, Electrospray Ionization , Tablets , Tandem Mass Spectrometry , Theophylline/metabolism , Uric Acid/analogs & derivatives , Uric Acid/urine , Xanthines/urineABSTRACT
To study the effect of Tibetan medicine Zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2, three different doses (1.2, 3.8 and 12 mg x kg(-1)) of Zuotai were administrated orally to rats once a day or once daily for twelve days, separately. Rats were administrated orally caffeine (CF) on the second day after Zuotai administration, and the urine concentration of CF metabolite 5-acetylamino-6-formylamino-3-methyl-uracil (AFMU), 1-methyluric acid (1U), 1-methylxanthine (1X), 1, 7-dimethylxanthine (17U) at 5 h after study drug administration was determined by RP-HPLC. The activity of CYP1A2 and NAT2 was evaluated by the ratio of metabolites (AFMU+1X+1U)/17U and the ratio of AFMU/(AFMU+1X+1U), respectively. The protein and mRNA expression of CYP1A2 and NAT2 were determined by ELISA and RT-PCR method, respectively. After single administration of Zuotai 3.8 mg x kg(-1) and repeated administration of Zuotai 3.8 and 12 mg x kg(-1), the activity of CYP1A2 and NAT2 decreased significantly compared with control group and there was no significant difference between other dose group and control group. The protein expression of CYP1A2 was significant lower than that in control group after repeated administration of Zuotai 12 mg x kg(-1), and the mRNA expression of CYP1A2 decreased significantly compared with that of control group after single administration of Zuotai 3.8 mg x kg(-1) and repeated admistration of Zuotai 12 mg x kg(-1), separately. The protein expression of NAT2 decreased significantly compared with that of control group after single and repeated administration of Zuotai 3.8 mg x kg(-1), respectively, and the mRNA expression of CYP1A2 decreased significantly compared with control group after single administration of Zuotai 3.8 mg x kg(-1). This study found that Tibetan medicine Zuotai had significant effect on the activity, protein and mRNA expression of CYP1A2 and NAT2.
Subject(s)
Arylamine N-Acetyltransferase/metabolism , Caffeine/metabolism , Cytochrome P-450 CYP1A2/metabolism , Drugs, Chinese Herbal/pharmacology , Medicine, Tibetan Traditional , Administration, Oral , Animals , Arylamine N-Acetyltransferase/genetics , Caffeine/urine , Cytochrome P-450 CYP1A2/genetics , Dose-Response Relationship, Drug , Drugs, Chinese Herbal/administration & dosage , Female , Male , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Theophylline/urine , Uracil/analogs & derivatives , Uracil/urine , Uric Acid/analogs & derivatives , Uric Acid/urine , Xanthines/urineABSTRACT
Phenotyping of cytochrome P450 2A6 (CYP2A6) was determined by assessing urinary caffeine metabolites in a Japanese population with a high frequency of CYP2A6 whole-gene deletion (CYP2A6*4). The levels of 1,7-dimethyluric acid (17U), 1-methylxanthine (1X), and 1,7-dimethylxanthine (17X) were measured in non-smokers whose CYP2A6 and NAT2 genotypes had been determined. Low 17U/1X ratios were observed in accumulated overnight urine samples of subjects genotyped as CYP2A6*4/*4 after caffeine treatment. The individual 17U/1X ratios in spot urine samples were almost constant before and 2-8 h after caffeine treatment, with or without prior abstention from dietary caffeine. The average 17U/1X ratios obtained from subjects with CYP2A6 *4/ *4 or CYP2A6 *1/ *4 genotypes were significantly lower than those from subjects with wild-type CYP2A6 *1/ *1 under dietary caffeine consumption. The present results suggest that impaired CYP2A6 function associated with CYP2A6 *4/ *4 could be determined using the 17U/1X ratios in spot urine samples under normal dietary caffeine consumption in Japanese non-smokers, without the need for additional caffeine administration or prior abstention from caffeine.
Subject(s)
Aryl Hydrocarbon Hydroxylases/genetics , Caffeine/metabolism , Theophylline/urine , Uric Acid/analogs & derivatives , Xanthines/urine , Adult , Caffeine/administration & dosage , Cytochrome P-450 CYP2A6 , Female , Gene Deletion , Genotype , Humans , Japan , Male , Phenotype , Pilot Projects , Uric Acid/urine , Young AdultABSTRACT
AIM: To evaluate the pharmacokinetic interactions between theophylline and antofloxacin in vivo and in vitro. METHODS: A randomized, 5-day treatment and 3-way crossover design was documented in 12 healthy subjects. The subjects were orally administered with antofloxacin (400 mg on d 1 and 200 mg on d 2 to 5), theophylline (100 mg twice a day and morning dose 200 mg on d 1 and 5), or theophylline plus antofloxacin. The plasma and urinary pharmacokinetics of antofloxacin and theophylline were characterized after the first and last dose. The effect of antofloxacin on theophylline metabolism was also investigated in pooled human liver microsomes. RESULTS: The 5-day treatment with antofloxacin significantly increased the area of the plasma concentration-time curve and peak plasma concentration of theophylline, accompanied by a decrease in the excretion of theophylline metabolites. On the contrary, theophylline did not affect the pharmacokinetics of antofloxacin. In vitro studies using pooled human hepatic microsomes demonstrated that antofloxacin was a weak reversible and mechanism-based inhibitor of CYP1A2. The clinical interaction between theophylline and antofloxacin was further validated by the in vitro results. CONCLUSION: The results showed that antofloxacin increases the plasma theophylline concentration, partly by acting as a mechanism-based inhibitor of CYP1A2.
Subject(s)
Anti-Bacterial Agents/pharmacology , Bronchodilator Agents/pharmacokinetics , Ofloxacin/analogs & derivatives , Theophylline/pharmacokinetics , Adult , Anti-Bacterial Agents/blood , Anti-Bacterial Agents/urine , Bronchodilator Agents/blood , Bronchodilator Agents/urine , Cross-Over Studies , Cytochrome P-450 CYP1A2/metabolism , Cytochrome P-450 CYP1A2 Inhibitors , Drug Interactions , Female , Humans , Male , Microsomes, Liver/metabolism , Ofloxacin/blood , Ofloxacin/pharmacology , Ofloxacin/urine , Theophylline/blood , Theophylline/urine , Young AdultABSTRACT
We developed a simple assay method for the determination of serum and urine norfloxacin and enoxacin using reversed-phase high-performance liquid chromatography and perchloric acid precipitation for sample pre-treatment. Optimized conditions can permit detection of norfloxacin and enoxacin in the same chromatogram, so either compound can be used as an internal standard for another determinant. Supernatants of the precipitated samples were analyzed by the octadecylsilyl silica-gel column under ambient temperature and an ultraviolet wavelength of 272 nm. A mobile phase solvent consisting of 20 mm sodium dihydrogenphosphate (pH 3.0) and acetonitrile (85:15, v/v) was pumped at a flow rate of 1.0 mL/min. The calibration curves for norfloxacin and enoxacin at a concentration of 62.5-1000 ng/mL for serum and 250-4000 ng/mL for urine were linear (r > 0.9997). The recoveries of norfloxacin and enoxacin from serum and urine were >94% with the coefficient of variations (CV) <5%. The CVs for intra- and inter-day assay of norfloxacin and enoxacin were <4.2 and <5.5%, respectively. This method can be applied to the pharmacokinetic study of norfloxacin and enoxacin after repeated administration to assess changes in CYP1A2 activity in healthy subjects.
Subject(s)
Chromatography, High Pressure Liquid/methods , Cytochrome P-450 CYP1A2 Inhibitors , Enoxacin/pharmacokinetics , Norfloxacin/pharmacokinetics , Adult , Caffeine/pharmacokinetics , Caffeine/urine , Chromatography, Reverse-Phase , Cytochrome P-450 CYP1A2 , Enoxacin/blood , Enoxacin/urine , Humans , Male , Norfloxacin/blood , Norfloxacin/urine , Reproducibility of Results , Sensitivity and Specificity , Theophylline/pharmacokinetics , Theophylline/urineABSTRACT
BACKGROUND: Widespread human exposure to phthalates, some of which are developmental and reproductive toxicants in experimental animals, raises concerns about potential human health risks. Underappreciated sources of exposure include phthalates in the polymers coating some oral medications. OBJECTIVE: The objective of this study was to evaluate whether users of phthalate-containing medications have higher urinary concentrations of phthalate metabolites than do nonusers. METHODS: We used publically available files from the National Health and Nutrition Examination Survey for the years 1999-2004. For certain survey periods, participants were asked to recall use of prescription medication during the past 30 days, and for a subsample of individuals, the urinary concentrations of phthalate metabolites were measured. We a priori identified medications potentially containing phthalates as inactive ingredients and then compared the mean urinary concentration of phthalate metabolites between users and nonusers of those medications. RESULTS: Of the 7,999 persons with information on urinary phthalate concentrations, 6 reported using mesalamine formulations, some of which may include dibutyl phthalate (DBP); the mean urinary concentration of monobutyl phthalate, the main DBP metabolite, among these mesalamine users was 50 times higher than the mean for nonusers (2,257 microg/L vs. 46 microg/L; p < 0.0001). Users of didanosine, omeprazole, and theophylline products, some of which may contain diethyl phthalate (DEP), had mean urinary concentrations of monoethyl phthalate, the main DEP metabolite, significantly higher than the mean for nonusers. CONCLUSION: Select medications might be a source of high exposure to some phthalates, one of which, DBP, shows adverse developmental and reproductive effects in laboratory animals. These results raise concern about potential human health risks, specifically among vulnerable segments of the general population and particularly pregnant women and children.
Subject(s)
Environmental Exposure/analysis , Pharmaceutical Preparations/administration & dosage , Pharmaceutical Preparations/analysis , Phthalic Acids/analysis , Adolescent , Adult , Data Collection , Didanosine/administration & dosage , Didanosine/analysis , Didanosine/urine , Environmental Exposure/statistics & numerical data , Female , Humans , Male , Mesalamine/administration & dosage , Mesalamine/analysis , Mesalamine/urine , Middle Aged , Omeprazole/administration & dosage , Omeprazole/analysis , Omeprazole/urine , Phthalic Acids/administration & dosage , Phthalic Acids/urine , Pregnancy , Theophylline/administration & dosage , Theophylline/analysis , Theophylline/urine , Young AdultABSTRACT
This study was performed to assess the influence of age, sex, smoking, and contraceptive use on CYP2A6 activity. In the metabolism of caffeine, the conversion of 1,7 dimethylxanthine (17X) to 1,7 dimethiylurate (17U) is catalyzed primarily by CYP2A6. CYP2A6 phenotype was determined by the urinary ratio 17U:17X in the interval of 4-5 h after caffeine intake in 179 healthy white Spaniards (102 women and 76 men). There were 99 non-smokers and 80 smokers. Among women, 26 were taking oral contraceptives. The age was the most important predictive factor of CYP2A6 activity (P < 0.001) with older subjects having higher activity. The influence of the gender was more modest (P = 0.07) with women exhibiting borderline increased values of the CYP2A6 marker than men. Tobacco smoking did not affect CYP2A6 activity. However, the CYP2A6 marker resulted to be strongly related to the use of oral contraceptives. The women users of oral contraceptives had higher values of CYP2A6 marker than both women not taking oral contraceptives and men (P < 0.001 in both comparisons). The results indicate that age, oral contraceptive use, and possibly gender should be controlled in epidemiological studies dealing with CYP2A6 activity and its relationship with xenobiotics exposure and genetic or pathological factor.
Subject(s)
Aryl Hydrocarbon Hydroxylases/metabolism , Contraceptives, Oral , Smoking , Adolescent , Adult , Age Factors , Biomarkers/urine , Caffeine/administration & dosage , Cytochrome P-450 CYP2A6 , Female , Humans , Male , Middle Aged , Phenotype , Risk Factors , Sex Factors , Spain , Theophylline/urine , Uric Acid/analogs & derivatives , Uric Acid/urineABSTRACT
A large number of metabolic alterations are increasingly being treated with growth hormone. Despite the fact that growth hormone is known to be the main regulator of several hepatic drug metabolizing enzymes in rodents, few studies deal with the effect of growth hormone on hepatic enzyme activities in human beings. The aim of this study was to determine the effects of growth hormone replacement therapy for 4 weeks on CYP2A6 activity in children, because changes in this enzyme activity may have important therapeutic and toxic consequences. A total of 31 growth hormone-deficient children (age range 4.1-13.1 years; mean age 9.88 +/- 2.89 years) participated. The genotypes of CYP2A6 gene, CYP2A6*1A, CYP2A6*1B, CYP2A6*4, CYP2A6*1x2 and CYP2A6*9, were determined by polymerase chain reaction. To assess the enzyme activity, we used caffeine as a probe drug at two points in time: before starting growth hormone therapy (Day 0) and after 4 weeks of growth hormone therapy (Day A). Caffeine and metabolite concentrations in urine were assayed by high-pressure liquid chromatography. The metabolite ratio 1,7-dimethilxanthine to 1,7-dimethylurate (17U/17X) served to indicate CYP2A6 activity. Median value and 95% confidence interval at baseline was 1.08 (0.98-1.24). The value after treatment was 1.08 (0.86-1.21). Data comparison between periods showed lack of statistically significant differences (P > 0.05). The relative change, measured by the ratio of medians and 90% confidence interval, was 1.02 (0.84-1.19). There were no significant differences when the ratio between genotype groups were compared. These results indicate that growth hormone replacement therapy of growth hormone-deficient children for 4 weeks does not modify the CYP2A6 activity and hence the efficacy or toxicity of the CYP2A6 substrate compounds.
Subject(s)
Aryl Hydrocarbon Hydroxylases/metabolism , Growth Disorders/drug therapy , Human Growth Hormone/deficiency , Human Growth Hormone/therapeutic use , Mixed Function Oxygenases/metabolism , Recombinant Proteins/therapeutic use , Adolescent , Aryl Hydrocarbon Hydroxylases/genetics , Caffeine/urine , Child , Child, Preschool , Chromatography, High Pressure Liquid , Cytochrome P-450 CYP2A6 , Female , Genotype , Growth Disorders/enzymology , Growth Disorders/genetics , Human Growth Hormone/administration & dosage , Humans , Injections, Subcutaneous , Male , Mixed Function Oxygenases/genetics , Recombinant Proteins/administration & dosage , Theophylline/urine , Uric Acid/analogs & derivatives , Uric Acid/urineABSTRACT
OBJECTIVE: To evaluate the pharmacokinetics of a brand of extended-release theophylline tablets and capsules in healthy cats. DESIGN: Randomized 3-way crossover study. ANIMALS: 6 healthy cats. PROCEDURES: A single dose of aminophylline (10 mg/kg [4.5 mg/lb], IV), a 100-mg extended-release theophylline tablet, or a 125-mg extended-release theophylline capsule was administered to all cats. Plasma samples were collected via preplaced central catheters throughout a 36-hour period. Plasma samples were frozen until analyzed by use of a fluorescence polarization monoclonal immunoassay. RESULTS: All cats tolerated drug administration and plasma collection with no adverse effects. Peak concentrations were reached for both orally administered products between 8 and 12 hours after administration. Bioavailability was excellent. Plasma concentrations were within the human therapeutic concentration of 5 to 20 microg/mL. CONCLUSIONS AND CLINICAL RELEVANCE: Daily administration of the brand of theophylline tablets and capsules used in this study at 15 mg/kg (6.8 mg/lb) and 19 mg/kg (8.6 mg/lb), respectively, maintained plasma concentrations within the desired therapeutic range in healthy cats.