An analysis of green discoloration of urine caused by propofol infusion.

BACKGROUND: Propofol is a short-acting, intravenous sedative-hypnotic agent that is widely used for the induction and maintenance of general anesthesia and sedation. An uncommon adverse effect of propofol is green discoloration of the urine, which has been reported not only under general anesthesia but also with sedation. Although it is assumed that the phenolic derivatives of propofol can cause green discoloration of the urine, the actual origin remains unknown. The aim of this report was to identify the origin of the green discoloration of the urine using liquid chromatography-mass spectrometry (LC-MS). CLINICAL FEATURES: The patient, a 51-year-old man, was scheduled for his oral surgery under general anesthesia using propofol. Postoperatively, the color of his urine was observed to be green. We compared and analyzed both the green urine and the normal urine using LC-MS. CONCLUSION: We experienced a case of a patient with green discoloration of the urine after general anesthesia using propofol. Although LC-MS analysis showed 2 unique peaks in the green urine at 490 and 590 nm, obvious causes were not revealed.

Grass-green urine from propofol infusion.

Green urine from propofol infusion is a benign and rare side effect. The discolouration appears when clearance of propofol exceeds hepatic elimination, and extrahepatic elimination of propofol occurs. This case report presents a 24-year-old male with grass green discolouration of urine based on propofol infusion.

Urinary propofol metabolites in early life after single intravenous bolus.

BACKGROUND: /st> Propofol clearance is lower in neonates than in adults and displays extensive interindividual variability, in part explained by postmenstrual age (PMA) and postnatal age (PNA). Since propofol is almost exclusively cleared metabolically, urinary propofol metabolites were determined in early life and compared with similar observations reported in adults. METHODS: /st> Twenty-four hours urine collections were sampled after a single i.v. bolus of propofol (3 mg kg(-1)) in neonates undergoing procedural sedation. Clinical characteristics (PMA, PNA, weight, and cardiopathy) were recorded. Urine metabolites [propofol glucuronide (PG), 1- and 4-quinol glucuronide (QG)] were quantified using high-pressure liquid chromatography. Urine recovery (% administered dose) and the contribution of PG and QG to urinary elimination were calculated. Data were reported by median and range, analysed by Mann-Whitney U or Spearman's rank. RESULTS: /st> Eleven neonates (median PNA 11 days, PMA 38 weeks) were included. Median propofol metabolite recovery was 64% (range 34-98%). PG contributed 34% (range 8-67%) and QG 65% (range 33-92%). There was no significant correlation between either PMA, PNA, or cardiopathy and propofol metabolites. Compared with adults, the contribution of PG (34% vs 77%) was lower and the contribution of QG (65% vs 22%) was higher in neonates. CONCLUSIONS: /st> Propofol metabolism in neonates differs from adults, reflecting the age-dependent limited glucuronidation capacity. Hydroxylation to quinol metabolites already contributes to propofol metabolism. These differences likely explain the PMA- and PNA-dependent reduced propofol clearance in neonates.

Recovery and long-term renal excretion of propofol, its glucuronide, and two di-isopropylquinol glucuronides after propofol infusion during surgery.

BACKGROUND: The metabolism of the short-acting anaesthetic agent propofol has been described over the first 24 h. However, the long-term disposition of propofol and its metabolites is unclear. We describe the pharmacokinetics (renal excretion rates and renal clearance) of propofol and its metabolites over 60 h. METHODS: Ten patients undergoing lung surgery were included in the study. They received anaesthesia with continuous i.v. propofol at an average rate of 10 mg min(-1). During surgery and 60 h thereafter, we sampled blood and urine. Propofol and its metabolites were measured using gradient high performance liquid chromatography (HPLC). RESULTS: In nine patients, propofol and its glucuronides were found in the plasma over the first 15 h. In the urine, however, even after 60 h, propofol and its quinol glucuronides were still detectable. One patient had a markedly different pharmacokinetic profile, showing a limited renal excretion or absorption of 12% of the dose. CONCLUSIONS: After an infusion of propofol, patients excrete propofol and its metabolites in the urine over a period in excess of 60 h. We hypothesize that (re)absorption of propofol and its metabolites by the kidney is a major process in elimination and that the reabsorbed compounds are gradually conjugated in the kidney and excreted in the urine. One patient showed a different pharmacokinetic profile for which we currently have no explanation.

Determination of thiopental in urine sample with high-performance liquid chromatography using iodine-azide reaction as a postcolumn detection system.

The reaction between iodine and azide ions induced by thiopental was utilized as a postcolumn reaction for chromatographic determination of thiopental. The method is based on the separation of thiopental on an Nova-Pak CN HP column with an acetonitrile-aqueous solution of sodium azide as a mobile phase, followed by spectrophotometric measurement of the residual iodine (lambda=350 nm) from the postcolumn iodine-azide reaction induced by thiopental after mixing an iodine solution containing iodide ions with the column effluent containing azide ions and thiopental. Chromatograms obtained for thiopental showed negative peaks as a result of the decrease in background absorbance. The detection limit (defined as S/N=3) was 20 nM (0.4 pmol injected amount) for thiopental. Calibration graphs, plotted as peak area versus concentrations, were linear from 40 nM. The elaborated method was applied to determine thiopental in urine samples. The detection limit (defined as S/N=3) was 0.025 nmol/ml urine. Calibration graphs, plotted as peak area versus concentrations, were linear from 0.05 nmol/ml urine. Authentic urine samples were analyzed, thiopental was determined at nmol/ml urine level.

Ultra-rapid procedure to test for gamma-hydroxybutyric acid in blood and urine by gas chromatography-mass spectrometry.

Gamma-hydroxybutyric acid (GHB) is a substance naturally present within mammal species. Properties of a neurotransmitter or neuromodulator are generally suggested for this substance. GHB is therapeutically used as an anaesthetic, but can be used for criminal offences (date-rape drug). It appears that the window of detection of GHB is very short in both blood and urine, and therefore its presence is very difficult to prove after a rape case. Twenty microl of blood or urine were pipetted into a glass tube, followed by 20 microl GHB-d(6) and 45 microl acetonitrile. After vortexing and efficient centrifugation, the supernatant was collected and evaporated to dryness. The residue was derivatized with BSTFA+1% TMCS for 20 min at 70 degrees C. After injection on a 30-m HP5 MS capillary column, GHB (m/z 233, 204 and 147) and GHB-d(6) (m/z 239) were identified by mass spectrometry. The procedure was linear from 1 to 200 mg/l for both blood and urine. Precisions were in the range 4 to 11%. The method appears simple, specific and rapid as an accurate result can be obtained within 1 h.

Investigation of propofol renal elimination by HPLC using supported liquid membrane procedure for sample preparation.

One of the least explored subjects in the research on the metabolism of a widely used anaesthetic, propofol, is its excretion in an unchanged form. According to literature, the estimated percentage of applied propofol eliminated intact via kidneys is lower than 0.3%. The present study shows the amount of propofol excreted in an unchanged form with urine collected during the first 48 h after anaesthesia in five patients undergoing elective intracranial procedures. The drug was concentrated and selectively isolated from urine samples by supported liquid membrane technique and determined by HPLC with fluorescence detection. The amount of unchanged propofol eliminated with urine was approximately (0.004 +/- 0.002)% of the total applied dose. The obtained results may suggest that propofol in an unchanged form is not excreted by kidneys at all provided that all propofol determined in presented study originated from conjugates hydrolysis.

Intra- and interindividual variations in urinary concentrations of endogenous gamma-hydroxybutyrate.

This study was designed to determine the urinary concentrations of endogenous GHB over a one-week period, the variations that occur within those concentrations, and whether those variations are affected by normalization to urinary creatinine. Its purpose was to ascertain whether endogenous concentrations fluctuate to such an extent that they may be misinterpreted as due to GHB ingestion. Every urine void produced by eight GHB-free subjects (five males and three females) over a one-week period was individually collected and analyzed for the presence of endogenous GHB and creatinine. The results of the non-normalized and normalized concentrations were statistically analyzed. Non-normalized GHB concentrations ranged from 0.00 to 6.63 microg/mL over seven days. The coefficients of variation (CV) for the individual non-normalized data were 44.0% to 77.7%. When the data were normalized to creatinine, the concentrations ranged from 0.00 to 6.79 microg/mg. The CVs for the creatinine-normalized results were between 29.7% and 76.8%. Analysis of the differences in CVs by the paired t-test (alpha = 0.05) found these improvements to be statistically insignificant. Such normalization allows for correction of urinary dilution or concentration by the kidneys which may affect endogenous GHB concentrations. The data also suggest significant (p < 0.001) differences in median endogenous urinary concentrations of GHB between males and females using the Mann-Whitney test. Because of the small number of subjects in this study, further investigations are required to substantiate this observation. Some of the subjects in this study demonstrated a strong tendency to produce higher or lower GHB concentrations at consistent periods during the day. This was most evident when looking at the creatinine-normalized concentrations. The results of our study indicate that there are significant intra- and interindividual variations in the urinary concentrations of endogenous GHB. Furthermore, there are also wide variations between individuals in the total daily amount of GHB excreted in the urine. Nonetheless, no specimen's GHB concentration approached 10 microg/mL (non-normalized) or 10 microg/mg (normalized). This study of the variability in endogenous urinary GHB excretion supports the recommendation of 10 microg/mL as an appropriate cutoff to identify exogenous GHB exposure in the absence of rare genetic deficiencies such as GHB aciduria. Patients with such a deficiency should be readily identifiable through prominent symptoms, repeated urinalysis, or genetic testing.

Propofol metabolites in man following propofol induction and maintenance.

BACKGROUND: The pharmacokinetics of propofol in man is characterized by a rapid metabolic clearance linked to glucuronidation of the parent drug to form the propofol-glucuronide (PG) and sulfo- and glucuro-conjugation of hydroxylated metabolite via cytochrome P450 to produce three other conjugates. The purpose of this study was to assess the urine metabolite profile of propofol following i.v. propofol anaesthesia in a Caucasian population. METHODS: The extent of phase I and phase II metabolism of propofol was studied in 18 female and 17 male patients after an anaesthesia induced and maintained for at least 4 h with propofol. The infusion rates (mg kg(-1) h(-1)) of propofol were (mean (SD)) 4.1 (1.0) and 4.5 (1.3) for males and females, respectively. Urine was collected from each patient for the periods 0-4, 4-8, 8-12, and 12-24 h after the start of propofol administration. In a preliminary study, the three main glucuro-conjugated metabolites were isolated from urine and characterized by magnetic resonance spectroscopy. The quantification of these metabolites for the different collection periods was then performed by a HPLC-UV assay. RESULTS: Total recovery of propofol in the metabolites studied amounts to 38%, of which 62% was via the PG metabolite and 38% via cytochrome P-450. This percentage is significantly higher than that previously reported from patients after a bolus dose of propofol. Extreme values for PG (0-24 h period) were included from 73 to 49%. There was no significant difference between female and male patients in the metabolite ratio. CONCLUSIONS: We conclude that the extent of hydroxylation in propofol metabolism was higher than in previous findings after administration of anaesthetic doses of propofol. Moreover, the ratio between hydroxylation and glucuronidation of propofol is subject to an inter-patient variability but this does not correlate with the dose of propofol. However, the variation of the metabolite profile observed in the present report does not seem to indicate an extended role of metabolism in pharmacokinetic variability.

Determination of gamma-hydroxybutyrate in water and human urine by solid phase microextraction-gas chromatography/quadrupole ion trap spectrometry.

A simple method of detection was developed for gamma-hydroxybutyrate (GHB). The method involves the derivatization of GHB using a hexyl-chloroformate procedure in aqueous media (such as water or urine), extraction of the derivatization product directly from the sample using solid-phase microextraction, and subsequent separation and detection with gas chromatography quadrupole ion trap mass spectrometry. The deuterated form of GHB (GHB-D6) is used as an internal standard for quantitation. The method was linear for GHB-spiked pure water samples from 2 to 150 microg/mL GHB with a detection limit of 0.2 microg/mL. Spiked urine samples showed linearity from 5 to 500 microg/mL GHB with a detection limit of 2 microg/mL. The SPME-GC/MS method is applied to actual case samples, and the results are compared to those values obtained using a conventional GC/MS method. Sensitivity and linearity are comparable to those seen using traditional methods of separation, yet the SPME method is superior due to the simplicity, speed of analysis, reduction in solvent waste, and ability to differentiate between GHB and gamma-butyrolactone (GBL).

Detection of new propofol metabolites in human urine using gas chromatography/mass spectrometry and liquid chromatography/mass spectrometry techniques.

Using hyphenated analytical techniques, gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS), a study on minor propofol metabolites in human urine was conducted. These techniques allowed identification of two new phase I metabolites (2-(omega-propanol)-6-isopropylphenol and 2-(omega-propanol)-6-isopropyl-1,4-quinol). In addition, their four corresponding conjugates (three glucuronides and one sulphate) were detected. Thus in human urine at least eight conjugate metabolites are produced, derived from four different aglycones (propofol; 2, 6-diisopropyl-1,4-quinol; 2-(omega-propanol)-6-isopropylphenol and 2-(omega-propanol)-6-isopropyl-1,4-quinol).

High-performance liquid chromatographic assay to detect hydroxylate and conjugate metabolites of propofol in human urine.

This paper describes a HPLC method for the simultaneous detection of phase I (2,6-diisopropyl-1-4-quinol and 2,6-diisopropyl-1-4-quinone) and phase II (4-(2,6-diisopropyl-1-4-quinol)-sulphate, 1-(2,6-diisopropyl-1-4-quinol)-glucuronide, 4-(2,6-diisopropyl-1-4-quinol)-glucuronide, and propofol-glucuronide) metabolites of propofol in human urine samples. Separation was based on a simple mobile phase and a reversed-phase chromatographic column. Metabolite identification was performed by UV spectrum on a diode-array detector and by LC-APCI-MS. The identification was also carried out using in vitro incubation mixtures (cytosol and microsomes prepared from liver) from several species: human, rat and rabbit. This assay was performed using UV, fluorescence and electrochemical detection modes. Each of these was analyzed and discussed.

Does use of propofol in heavy alcohol drinkers tend to discolor their urine?

A case in which the urine of a patient who was a heavy drinker turned pink when propofol was used for anesthesia is reported, and a new cause of the urine discoloration is proposed.

Direct high-performance liquid chromatography determination of propofol and its metabolite quinol with their glucuronide conjugates and preliminary pharmacokinetics in plasma and urine of man.

Propofol (P) is metabolized in humans by oxidation to 1,4-di-isopropylquinol (Q). P and Q are in turn conjugated with glucuronic acid to the respective glucuronides, propofol glucuronide (Pgluc), quinol-1-glucuronide (Q1G) and quinol-4-glucuronide (Q4G). Propofol and quinol with their glucuronide conjugates can be measured directly by gradient high-performance liquid chromatographic analysis without enzymic hydrolysis. The glucuronide conjugates were isolated by preparative HPLC from human urine samples. The glucuronides of P and Q were present in plasma and urine, P and Q were present in plasma, but not in urine. Quinol in plasma was present in the oxidised form, the quinone. Calibration curves of the respective glucuronides were constructed by enzymic deconjugation of isolated samples containing different concentrations of the glucuronides. The limit of quantitation of P and quinone in plasma are respectively 0.119 and 0.138 microg/ml. The limit of quantitation of the glucuronides in plasma are respectively: Pgluc 0.370 microg/ml, Q1G 1.02 microg/ml and Q4G 0.278 microg/ml. The corresponding values in urine are: Pgluc 0.264 microg/ml, Q1G 0.731 microg/ml and Q4G 0.199 microg/ml. A pharmacokinetic profile of P with its metabolites is shown, and some preliminary pharmacokinetic parameters of P and Q glucuronides are given.

Anodic adsorptive stripping voltammetric determination of the anesthetic drug: methohexital sodium.

Methohexital (MS) determination is based on the formation of insoluble mercury salt on a hanging mercury drop electrode after preaccumulation by adsorption. This property was exploited in developing a highly sensitive stripping voltammetric procedure for the determination of the drug. The anodic current of adsorbed compound is measured by linear sweep anodic stripping voltammetry (LSASV), preceded by a period of preconcentration. The effect of various parameters such as supporting electrolyte composition, pH, initial potential, scan rate, accumulation time and ionic strength are discussed to characterize the interfacial and redox behavior. The detection limit was found to be 2x10(-7) M (56.8 ppb) with 180-s accumulation time. The interference of some amino acids, ascorbic acid and some metal ions was investigated. The application of this method was tested in the determination of methohexital in spiked urine samples. The precision of the method is satisfactory with a relative standard deviation of 2.5%.

Gas chromatographic-mass spectrometry and gas chromatographic-Fourier transform infrared spectroscopy assay for the simultaneous identification of fentanyl metabolites.

Fentanyl, a synthetic opioid, undergoes important biotransformation to several metabolites. A gas chromatographic-mass spectrometric assay was applied for the simultaneous analysis of fentanyl and its major metabolites in biological samples. The identification of different metabolites was performed by gas chromatography-mass spectrometry (electronic impact and chemical ionisation modes) and gas chromatography-Fourier transform infrared spectroscopy. In the present study, rat and human microsomes incubation mixtures and human urines were analysed. In vitro formation of already known fentanyl metabolites was confirmed. The presence of metabolites not previously detected in human urine is described.

Pharmacokinetics of thiopentone enantiomers following intravenous injection or prolonged infusion of rac-thiopentone.

AIMS: Thiopentone is administered as a racemate (rac-thiopentone) for induction of anaesthesia as well as for neurological and neurosurgical emergencies. The pharmacokinetics and pharmacodynamics of rac-thiopentone have been extensively studied but the component R-(+)- and S-(-)- enantiomers, until very recently, have been largely ignored. METHODS: The present study analyses the pharmacokinetics of R-(+)- and S-(-)-thiopentone in 12 patients given rac-thiopentone intravenously for induction of anaesthesia and five patients given a prolonged infusion of rac-thiopentone used for treatment of intracranial hypertension. RESULTS: The mean total body clearance (CLT) and apparent volume of distribution at steady-state (Vss) showed trends towards higher values for R-(+)- than for S-(-)-thiopentone in both patient groups; CLT and Vss of unbound fractions of R-(+)- and S-(-)-thiopentone, however, did not show these trends. The time courses of R-(+)- and S-(-)- thiopentone serum concentrations were so similar that EEG effect could not be attributed to one or other enantiomer. Serum protein binding for S-(-)-thiopentone was greater than for R-(+)-thiopentone (P = 0.02) and 24 h urinary excretion of R-(+)-thiopentone was greater than for S-(-)-thiopentone (P = 0.03). In one patient, concomitant measurement of CSF and serum thiopentone concentrations found that serum: CSF equilibration of unbound fractions of both enantiomers was essentially complete. CONCLUSIONS: The study was unable to determine any pharmacokinetic difference of clinical significance between the R-(+)- and S-(-)-thiopentone enantiomers and concludes that minor differences in CLT and Vss could be explained by enantioselective difference found in serum protein binding.

Determination of diprivan in urine by a supported liquid membrane technique and liquid chromatography-electrochemical detection.

A supported liquid membrane technique was used for the extraction and enrichment of propofol in a spiked sample of urine. An acidic solution of propofol and thymol as an internal standard was passed over the membrane and after enrichment the acceptor solution was analyzed by LC with an electrochemical detector. The acceptor and donor pH, flow-rate, and volume of donor and different membrane solvents were varied to optimize the extraction efficiency. The detection limit for 100 ml of a spiked urine sample was 10 ppt of propofol.