Recovery Of Morphine Research Articles

Reducing harm through the development of good preparation practices for the injection of slow release morphine sulphate capsules

BackgroundIt is not always easy to advise people who inject drugs (PWID) on how to prepare their drugs in a way that is associated with reduced harm. This is particularly true for pharmaceutical drugs that are not meant to be injected. Our objective was to find “good preparation practices” for slow release morphine sulphate capsules, namely preparation methods that reduce harm, that are evidence-based and acceptable to PWID.MethodsIn the laboratory, morphine sulphate capsules were prepared using both a cold and lukewarm preparation technique, two contact and stirring durations (1 min and 20 s) and 4 different filters (cotton filter, Sterifilt, Sterifilt+ and a wheel filter). The following outcomes were compared: particle reduction and morphine content in the filtrate, as well as filtration ease and time.ResultsThe lukewarm method and a stirring and contact time of 1 min were associated with a considerably higher morphine yield than both the cold method and the stirring time of only 20 s. Moreover, the suspension obtained was easy to filter using membrane filters. Particle reduction was important with all three membrane filters tested. Using the lukewarm method, morphine recovery was 86% for the wheel filter, 89% for the Sterifilt and 99% for the Sterifilt+.ConclusionsThe provision of a method that is easy to use, reduces harms associated to the injection of insoluble particles and recovers virtually all the active drug has a large chance to be adopted by people who use drugs. This type of “best practices” can be provided by drug workers and by people who use drugs to actively promote harm reduction.

Analysis of opioids in postmortem urine samples by dispersive liquid-liquid microextraction and high performance liquid chromatography with photo diode array detection

BackgroundOpioids abuse and related deaths are increasing in the world. Therefore, the design of new analytical methods for detection of opioids in biological samples is necessary for clinical and forensic settings.MethodsIn this study, dispersive liquid-liquid microextraction (DLLME) combined with high-performance liquid chromatography with photo diode array detector (HPLC-PDA), as a new and sensitive method were examined for the extraction and determination of morphine, codeine and methadone in postmortem urine samples. Effective factors on DLLME were optimized. The extracts were analyzed by HPLC-PDA using a Eurospher® C18 column (250 mm × 4.6 mm, particle size: 5 μm).ResultsThe volumes of chloroform as the extraction solvent and acetone as the dispersive solvent were selected 300 μl and 500 μl, respectively. The optimum pH 9.8 and extraction time was 0.5 min were selected. Under optimum condition, the enrichment factor and the recovery of morphine, codeine, and methadone spiked into postmortem urine samples were in the range of 175–215.8 and 87.5–107.9%, respectively. Calibration curves for each analyte are linear in the range of 0.5–100 μg ml− 1. Limit of detection (LOD) for the analytes was in the range of 10–25 μg l− 1. Finally, the proposed method was successfully applied to 50 postmortem urine samples for determination of the opioids.ConclusionsThe proposed method is an easy, fast, low cost and efficient for the extraction and determination of opioids in postmortem urine samples and should be considered as analytical method for determination of opioids in forensic and clinical toxicology labs.

Electrocatalytic Behavior and Amperometric Detection of Morphine on ITO Electrode Modified with Directly Electrodeposited Gold Nanoparticles

AbstractA gold nanoparticles (AuNPs) modified indium tin oxide (ITO) film coated glass electrode was prepared via a novel electrochemical deposition technique. The UV‐visible spectrum and SEM indicated that the AuNPs on ITO electrode surface were spherical shape and quite symmetric distributed. The modified electrode exhibited excellent catalytic activity for the oxidation of morphine (MO). At optimal experimental condition, the oxidation current was responsive with the MO concentrations ranging from 8.0×10−7 to 1.6×10−5 M, the detection limit was 2.1×10 –7 M. The modified electrode also exhibited high stability and reproducibility. The average recoveries of detection MO in human urine were ranged between 91.95% and 92.23%, and the RSD was less than 3.68% (n=5).

A Rapid and Reliable Solid-Phase Extraction Method for High-Performance Liquid Chromatographic Analysis of Opium Alkaloids from Papaver Plants

A rapid and reliable solid-phase extraction method for HPLC analysis of opium alkaloids from Papaver plants was established. Fifty mg of dried and powdered plant sample was extracted with 5 ml of 5% acetic acid for 30 min under sonication. After centrifugation, 3 ml of the supernatant was loaded on a reversed-phase cation-exchange solid-phase extraction cartridge. After seriate washings with 0.1 M hydrochloric acid and methanol, alkaloids were eluted with a mixture of 28% ammonia and methanol (1:19). The eluate was concentrated under nitrogen stream at 40 degrees C and the residue was dissolved in 50% aqueous methanol for high performance liquid chromatographic analysis. With this solid-phase extraction method, the recovery of morphine, codeine, oripavine, thebaine, papaverine, noscapine and sanguinarine was from 99.94 to 112.18% when the standard alkaloids were added to the plant samples. Opium alkaloids of a variety of genus Papaver plants cultivated in a field and phytotron were analyzed by this method.

Determination of Morphine by Capillary Zone Electrophoresis Immunoassay Combined with Laser‐Induced Fluorescence Detection

A competitive immunoassay for detecting morphine in bio‐samples was established by capillary zone electrophoresis combined with laser‐induced fluorescence detection (CZE‐LIF). The antigen of morphine was labeled with isothiocyano‐fluorescein (FITC) and then incubated with morphine monoclonal antibody and samples. The linear range was 50–1000 ng/mL, which was suitable for clinical and forensic applications. The detection limit can reach 40 ng/mL, based on S/N = 2. The recoveries of morphine from serum were satisfactory.

Determination of morphine and morphine glucuronides in human plasma by 96-well plate solid-phase extraction and liquid chromatography–electrospray ionization mass spectrometry

There is considerable interest in quantifying morphine and its major metabolites, morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G). Available assays use gas chromatography–mass spectrometry or high-performance liquid chromatography (HPLC) with single or tandem mass spectrometry, ultraviolet, electrochemical, or fluorimetric detection. Nevertheless, few methods provide adequate sensitivity for all analytes, in a single injection, with the desired rate of sample throughput. A rapid and sensitive method for quantification of morphine, M3G and M6G from human plasma using HPLC with electrospray ionization mass spectrometry was developed using a Waters Oasis MCX 96-well plate for extracting both lipophilic morphine and its hydrophilic glucuronides, C18 separation using an isocratic mobile phase (methanol, acetonitrile and formic acid), and selected ion monitoring. Recoveries of morphine, M3G and M6G, respectively, were 81, 90 and 82% at the low (2, 25 and 2 ng/ml), 80, 77 and 75% at the medium (10, 250 and 10 ng/ml), and 74, 62 and 72% at the high (100, 1000 and 100 ng/ml) quality control samples. The limit of quantitation was 0.5 ng/ml morphine and M6G, and 5 ng/ml M3G. Analytes were validated over a linear range of 0.5–200 ng/ml morphine and M6G, and 5–2000 ng/ml M3G. This assay represents an improvement over existing methods through solid phase extraction with increased sample throughput (96-well plates), use of small samples (0.5 ml), and sub-nanogram detection.

Determination of morphine in urine by solid-phase immunoextraction and high-performance liquid chromatography with electrochemical detection

The analysis of morphine in biological fluids is of vital interest in monitoring opiate abuse and in drug abuse research. Although methods for analysis of morphine and its metabolites are well established, studies are still being carried out to improve sample preparation procedures as well as detection levels of morphine in biological samples. In this study, morphine-specific immunosorbents were developed to concentrate morphine prior to HPLC analysis. Urine (0.1 ml) was diluted 10-fold with phosphate-buffered saline, pH 7.4 (PBS), loaded onto a solid-phase immunoextraction column and washed with 15 ml PBS followed by elution with 2 ml of elution buffer (40% ethanol in PBS, pH 4). The eluted fraction was analysed for morphine by HPLC–electrochemical detection using a cyanopropyl (CN) analytical column with 25% acetonitrile in phosphate buffer–sodium lauryl sulphate, pH 2.4 as the mobile phase. Duration of the extraction procedure was approximately 40 min. Calibration graphs were linear from 100 ngml−1 to 500 ngml−1 in urine. The inter-assay R.S.D. was <10% and the recovery of morphine from urine was >98%. Immunocolumns demonstrated remarkably high specificity towards morphine showing minimal binding with other opiate metabolites such as codeine, normorphine, norcodeine, morphine-3-glucuronide, morphine-6-glucuronide.

A rapid and sensitive quantitation method of endogenous morphine in human plasma.

Endogenous morphine is found at very low levels (pg/ml) in human plasma. Without technical modification, the usual techniques, i.e., RIA or HPLC, can not directly detect its presence. As a result the initial purification process is very crucial. In this report we demonstrate this opiate alkaloids determination in 2 ml of human plasma. Following an acidified precipitation of proteins in plasma, a solid-phase extraction, Sep-pak collection, was employed to concentrate the endogenous morphine. The extract was dissolved in a pH 8.9 phosphate buffer and then extracted into an organic solvent (chloroform and isopropyl alcohol). The initially purified sample was separated by HPLC and detected with an electrochemical detector of 500 mV and 0.02 Hz. The results showed that the recovery of morphine by this technique was 86+5.7%, and the sensitivity was 50 pg/ml. Compared to the detection sensitivity of the HPLC (2 ng/ml), this method increased the detection limit by 40-fold. With this method the concentration of endogenous morphine in plasma was determined as 80 pg/ml.

Biotransformation and pharmacokinetics of ethylmorphine after a single oral dose.

1. The pharmacokinetics of ethylmorphine after administration of a single dose of the cough mixture Cosylan were investigated in 10 healthy subjects. 2. The median urinary recovery of ethylmorphine and measured metabolites was 77% over 48 h. The median tmax of unchanged ethylmorphine was 45 min, and the terminal elimination t1/2 was 2 h. Ethylmorphine-6-glucuronide was found to be the major metabolite. 3. Two subjects had significantly lower urinary recovery (0.48 h) of morphine and morphine-glucuronides than the remainder. Furthermore, these two had urinary metabolic ratios (MRO) and partial metabolic clearances (CLmO) for O-deethylation of ethylmorphine tentatively classifying them phenotypically as poor metabolisers of the debrisoquine/sparteine type. 4. Genotyping for cytochrome P450 (CYP) 2D6 alleles revealed five homozygote (wt/wt) and five heterozygote subjects. Two subjects phenotypically classified as poor metabolisers were genotypically CYP2D6A/wt and CYP2D6D/wt, respectively. 5. Serum and urine samples taken more than 8 and 24 h after administration of ethyl-morphine respectively, contained morphine and morphine-glucuronides, but no ethylmorphine, ethylmorphine-6-glucuronide or (serum only) norethylmorphine. Norethylmorphine could be detected after hydrolysis of urine samples in all subjects. The urinary recovery of the active metabolites morphine and morphine-6-glucuronide after administration of ethylmorphine varied by a factor of 9 between individuals. 6. The wide variation in recovery of morphine and morphine-glucuronides after oral administration of ethylmorphine could not be explained simply by a difference in CYP2D6 genotype. Constitutional variation in other enzymatic pathways involved in ethylmorphine metabolism is probably crucial. Ratios of morphine to parent drug cannot be used to distinguish the source of morphine after administration of ethylmorphine. Norethylmorphine should be included in urine assays for opiates in forensic toxicology, and no firm conclusions about the source of morphine are possible based on serum samples obtained more than 24 h after drug administration.

Lacrimal and Plasmatic Kinetics of Morphine After an Ophthalmic Delivery of Three Different Formulations

AbstractThe aims of this work were to study the influence of a thermoreversible gel and an insert containing morphine on lacrimal and plasmatic kinetics in comparison with a simple solution. Lacrimal uptake was carried out with a Millipore filter which was characterized by a total recovery of morphine and an absence of inter. ference on the CPG/SM procedure.Contrary to the thermoreversible gel, the inserts prolonged the lacrimal and plasmatic kinetics. In the lacrimal fluid, Cmax was reduced from 21.9 to 4.8 mg/ml and Tmax delayed from 2.9 to 51.4 minutes. Likewise, in the plasma, Cmax was decreased from 116 to 37 ng/ml and Tmax prolonged from 17 to 135 minutes. This plasmatic prolongation was equivalent to the one reported with oral prolonged release devices containing morphine. Moreover, the insert areas under the curve (taking account of the delivered dose) in the lacrimal fluid and in the plasma were multiplied by from 1.6 and 2.6, respectively, in comparison with the solution.

Comparative Study of Different Extractive Procedures to Quantify Morphine in Urine by HPLC-UV

Abstract A comparative study to extract and purify total morphine in urine is described using several procedure treatments of biological samples. Sep-Pak C18 cartridges, Extrelut columns and liquid-liquid conventional extraction with sequencial purification through Extrelut column were assayed in order to establish the best procedure to quantitatively extract morphine from urine free of endogenous interferents. Analysis of the drug was made by HPLC-UV with a C18 column and methanol: acetate buffer (pH6.9) (70+30) as eluent. Recovery of morphine from spiked urine was 88.0% and 79.8% for concentrations of 1μg/ml and 5μg/ml, respectively. The detection limit of the method was 0.2μg/ml and coeficient of variation was 1.8% (n+4).

Sensitive and specific radioimmunoassays for opiates using commercially available materials. I: Methods for the determinations of morphine and hydromorphone

Sensitive and specific radioimmunoassay (RIA) methods for the analysis of morphine and hydromorphone in human plasma samples using commercially available materials were developed. The limit of quantitation was 0.3 ng/mL of plasma for morphine and 50 pg/mL of plasma for hydromorphone. An extraction step preceding the RIA quantitatively removed morphine, leaving 99% morphine-3-glucuronide (M3G) and 95% hydromorphone-3-glucuronide (H3G) in the aqueous phase. The specificity of the morphine RIA method for the analysis of clinical samples was confirmed by HPLC quantitation. The antiserum used in the hydromorphone RIA method cross reacted slightly with H3G at 0.66%. However, analysis of clinical samples using the direct versus the extraction RIA showed that the extraction step was necessary for the specific determination of hydromorphone in pharmacokinetic studies. After extraction, only 0.033% of the H3G present in plasma samples would be observed as interference for the free hydromorphone. The RIA methods were shown to be accurate and reproducible with almost 100% recovery of morphine and hydromorphone. They offer convenient alternatives to chromatographic methods for pharmacokinetic studies.

Voltammetric behaviour of morphine at a glassy carbon electrode and its determination in human serum by liquid chromatography with electrochemical detection under basic conditions

The electrochemical oxidation of morphine was studied at pH values of between 7.00 and 12.00 by cyclic voltammetry and chronoamperometry at a planar glassy carbon electrode. The peak potential was dependent on pH over the range 7.00-9.75; it was independent of pH above the latter value, indicating a pKa value of 9.75. The peak current was found to be independent of pH, ionic strength of phosphate buffer (0.02-0.1 mol dm-3) and percentage of acetonitrile (0-40% v/v). The oxidation was found to occur in three steps; these are considered to result from a one-electron oxidation of the phenoxide group, followed by a one-electron loss from the oxidation product, pseudomorphine, and finally a two-electron loss from a tertiary amine group. A simple method of analysis by high-performance liquid chromatography was developed which employed a column packed with a reversed-phase, pH-stable, octadecylsilane-modified silica. Separation was achieved with a mobile phase containing 20% v/v acetonitrile in 0.05 mol dm-3 phosphate buffer, pH 11.0. Amperometric detection was carried out with an applied potential of +0.45 V versus Ag-AgCl. The detection limit was 1.24 x 10(-13) mol of morphine injected. The detector gave a linear response from 1.2 x 10(-12) to 4.0 x 10(-10) mol of morphine injected. The extraction method required 0.5 ml of serum, and no solvent evaporation was needed. The recovery of morphine was 80.9%. The method gave a linear response to at least 15.0 x 10(-7) mol dm-3.(ABSTRACT TRUNCATED AT 250 WORDS)

Recovery of morphine from a controlled-release preparation: A source of opioid abuse

MS-Contin (Purdue-Fredrick, Norwalk, CT) is a controlled-release preparation of morphine sulfate that has demonstrated efficacy in the management of chronic cancer pain problems. It has recently come to the attention of the authors that MS-Contin represents a potential source of opioid abuse through the extraction and intravenous injection of the morphine from this preparation. The authors describe a simple aqueous extraction method that was used to quantitatively determine the extent to which morphine could be obtained from MS-Contin tablets.

Rapid quantitative analysis of morphine in blood by flash-heater methylation and gas chromatography with a nitrogen-specific detector

A simplified gas chromatographic method for the rapid determination of morphinn in rat and human blood is described. After single extraction, morphine was reconstituded in trimethylanilinium hydroxide and introduced directly into a gas chromatograph equipped with a nitrogen specific flame ionization detector. Limits of detection were 0.5μg/ml for either water or blood solutions. Analytical recoveries of morphine, added to either water or blood were 88±6 and 90±3% $$(\bar x \pm SD)$$ , respectively, when compared with morphine dissolved in methanol. The between-run coefficients of variation were 5.0 and 5.4% for water and blood, respectively, over the range of 0.5–32.2μg/ml, whereas the within-run coefficients of variation at 1μg/ml were 8.1 and 7.9%, respectively. The application of the method to an in vivo animal experiment is demonstrated.

Time-course of alterations in morphine-induced analgesia and nociceptive threshold following medullary raphe lesions

Electrolytic lesions which included major portions of the nucleus raphe magnus produced alterations in nociceptive threshold and morphine-induced analgesia. These effects were highly dependent on the time at which measurements were made following the lesioning procedure. Seven days or less after placement of the lesions there was a pronounced decrease in nociceptive threshold, but no effect on the capacity of morphine to induce antinociception. However, the nociceptive threshold exhibited recovery by 21–28 days, during which time there was a gradual, but profound, decrease in the capacity of morphine to induce analgesia. At longer times after the placement of lesions (28–35 days), the nociceptive threshold, assessed using the tail-flick test, became elevated concomitant with a recovery of morphine's analgesic potency. The differential time course of the lesion-induced attenuation of morphine analgesia and decrease in nociceptive threshold suggests that these effects result from the destruction of separate neuronal systems. Thus, the lesion-induced decrease in nociceptive threshold probably results from destruction of raphespinal serotonergic neurons while the attenuation of morphine analgesia is most likely an indirect result of damage to non-serotoninergic raphe-spinal neurons. Finally, the failure of acute raphe lesions to alter morphine analgesia indicates that the neurons found in the raphe magnus are not directly involved in mediating the antinociceptive actions of opiates.

Detection of Drugs Using XAD-2 Resin. III: A Routine Screening Procedure for Bile

The ability of bile to concentrate drugs and metabolites coupled with its general availability make it suitable for analysis and often the fluid of choice in postmorten cases requiring drug screening. Bile (5 to 10 ml) was diluted with water, sulfuric acid was added, and the mixture was autoclaved. The precipitated bile salts were easily removed by filtration and the filtrate (pH adjusted to 8.0 to 8.5) extracted with XAD-2 resin. Drugs were eluted with a mixture of ethyl acetate/1,2-dichloroethane and analyzed with thin-layer chromatography. Varying the dilution of bile improved the recovery of morphine, codeine, methadone, amobarbital, and phenobarbital. Excessive dilution, however, caused a washing phenomenon and reduced recovery of some drugs, as shown with morphine and codeine. The procedure described is useful for the rapid screening of bile specimens for drugs.

Recovery of Morphine from Biological Samples by Hydrolysis and Solvent Extraction

Abstract Quantitative recovery of drugs from biological samples is important when a response is to be related to the amount of drug present in a tissue sample. Morphine is one drug of interest in this regard because of its widespread use and its chemical peculiarities. Its relatively low dosage, amphoteric nature, and metabolism to a water-soluble product, 3-morphine monoglucuronide [1,2], make morphine relatively difficult to analyze in biological samples. Most quantitative analytical schemes of analysis require that the glucuronide be cleaved to free morphine for extraction into an organic solvent. Acid hydrolysis [3] and enzymatic cleavage [4] are the most popular methods for freeing the morphine. We report here a study of the recoveries of radioactively tagged morphine from biological samples of morphine-treated dogs by using hydrolysis and solvent extraction. It is possible to recover more than 90% of the morphine contained in a sample with acid hydrolysis and about 80% with enzymatic cleavage.

Improved recovery of morphine from biological tissues using siliconized glassware.

The adsorption of morphine onto glassware during analysis in aqueous solutions and biological tissues is a common problem, resulting in lower recoveries than expected. The problem can be remedied by siliconization of all glassware involved. The same glassware cannot be used repeatedly without incurring a significant loss of morphine.

Studies on the Use of XAD-2 Resin for Detection of Abused Drugs in Urine

Abstract We describe a procedure for extracting weakly acidic, neutral, and basic drugs from urine by using a column of XAD-2 resin. Adsorption of drugs from 20 ml of urine buffered at pH 8.5 ± 0.5 at a controlled flow rate of 2.5 ml/min was greater than 89% for all drugs tested except aspirin. On eluting the drugs tested with acetone and methanol/chloroform, recoveries ranged from 75 to 93%. Overall recoveries of drugs from urine to a thin-layer chromatography plate were between 63 and 78%. The concentration of morphine added to normal urine that can be detected 99% of the time (95% confidence limits) by this method was 80 (65-100) µg/liter. We evaluated three methods for recovering morphine from morphine glucuronide added to urine, by using appropriate modifications of the XAD-2 resin extraction method. Hydrolysis of urine, hydrolysis of urine extracts adsorbed on XAD-2 resin, and hydrolysis of urine extracts from the XAD-2 resin followed by a solvent extraction gave 75%, 40%, and 10% recoveries of morphine, respectively.