Gas chromatographic/mass spectrometric analysis of morphine and codeine in human urine of poppy seed eaters.

In this study, poppy seeds were examined for a natural constituent that might serve as a maker for the seeds' ingestion as opposed to opiate abuse. Thebaine was selected as possible marker, since it was found to be a component of all poppy seeds examined and was not a natural component of different heroin samples. During the course of this investigation, a new extraction and cleanup procedure was developed for the gas chromatographic/nitrogen phosphorus detection (GC/NPD) and gas chromatographic/mass spectrometric (GC/MS) analysis of morphine and codeine in urine. A linear response, over a concentration range of 25 to 600 ng/mL, was obtained for codeine and morphine (r = 0.9982 and 0.9947, respectively). The minimum detectable level (LOD) and limit of quantitation (LOQ) for morphine were 10 and 30 ng/mL, respectively; whereas LOD and LOQ for codeine were 2 and 8 ng/mL, respectively. The coefficients of variance (CV, n = 6) for morphine and codeine analyses at the 100-ng/mL level were 13.3 and 4.6%, respectively. This procedure was used for the analysis of urine samples from five poppy seed eaters who each ingested 200 g of poppy seed cake. Results indicated that significant amounts of morphine and codeine are excreted in urine and that in all subjects, at least at one point in time, the apparent morphine concentration as determined by radioimmunoassay (RIA) analysis exceeded the cutoff value (300 ng/mL) established for screening. Thebaine was not detected in urine specimens collected following poppy seeds ingestion and thus could not be used as a marker.

Separation of opium alkaloids by carbon dioxide sub- and supercritical fluid chromatography with packed columns. Application to the quantitative analysis of poppy straw extracts.

The optimization of the separation of seven opium alkaloids by sub- and supercritical fluid chromatography (SFC) using packed columns and carbon dioxide as the primary mobile phase was studied. The influence of aminated polar modifiers on bare and aminopropyl-bonded silica was investigated. It was found that the presence of an amine in the mobile phase could enhance the retention of alkaloids on aminopropylsilica. Various separation schemes were possible depending on the type of analysis needed. An aminopropyl-bonded silica used with a carbon dioxide-methanol-triethylamine-water mixture (82.95:16.25:0.50:0.30, w/w) gave a very rapid separation (2 min). A bare silica with a carbon dioxide-methanol-methylamine-water mixture (83.37:16.25:0.15:0.23, w/w) gave longer analysis (10 min) but a higher resolution. This last procedure was applied to a poppy straw extract and to the determination of three alkaloids of interest after a peak purity study using a diode-array UV detector. For these alkaloids, SFC appears to be a promising technique for the routine analysis of opium alkaloids.

Fatal intoxications in Denmark following intake of morphine from opium poppies.

In Denmark it is legal to grow opium poppies for the production of poppy seeds and until 1986 for decoration purposes, too. Danish poppy capsules contain 0.3-5 mg morphine per capsule and the content of morphine in opium exuded from the capsules may amount to 24%. This has resulted in misuse as both fresh and dried poppy capsules have been used for the production of "opium tea". During the period 1982-1985, seven casualties occurred among drug addicts in Denmark which were solely or partly caused by these opium poppies.

Development of fluoroimmunoassays for the specific detection of morphine in urine.

Two fluoroimmunoassays for the specific detection of morphine in urine are described based on the use of ovine antibodies and fluorescein-labelled normorphine. The first, a polarisation fluoroimmunoassay, is performed by adding 10 microliter of urine to 1.5 ml of a single-reagent, comprising premixed antiserum and tracer, incubation for a few minutes at ambient temperature and measurement of fluorescence polarisation. The assay gives results which compare well with those by thin-layer chromatography, EMIT d.a.u., and the Boehringer opiate drug test. Although adequate for routine screening for drug abuse, the technique is not as sensitive as some radioimmunoassays. Therefore, a second fluoroimmunoassay was developed based on the use of the same antibodies covalently coupled to magnetisable particles to facilitate both the separation of the bound and free fractions and the removal of non-specific interfering substances. Thus, larger sample volumes could be employed and greater sensitivity achieved.

Opiates in poppy seed: effect on urinalysis results after consumption of poppy seed cake-filling.

We report the analysis of poppy seed filling for morphine and codeine content. Concentrations in the range 17.4 to 18.6 micrograms/g (morphine) and 2.3 to 2.5 micrograms/g (codeine) were found in different lots of the filling, which is widely used in baking. The effect of consumption of poppy seed filling on opiate urinalysis results is discussed. Morphine concentrations as high as 4.5 mg/L are reported, with persistence of concentrations greater than 0.3 mg/L as long as 35 h after consumption.

Concentrations of morphine and codeine in serum and urine after ingestion of poppy seeds.

We measured morphine and codeine in commercially available poppy seeds and in serum and urine samples from healthy adults who had ingested these poppy seeds. Four brands of black poppy seeds, examined by gas chromatography-mass spectroscopy (GC-MS) with deuterated internal standards, contained from 17 to 294 micrograms of morphine and 3 to 14 micrograms of codeine per gram of seeds. Morphine was detected by GC-MS in hydrolysates of serum as late as 24 h after ingestion, with a maximum mean concentration of 100 ng/mL (range 82-131) measured 2 h after the subjects ingested 25 g of seeds. Opiates were detectable (greater than 300 micrograms/L) in urine by enzyme-multiplied immunoassay (EMIT; Syva Co.) and by radioimmunoassay screening procedures for as long as 48 h after ingestion. The identity and quantities of morphine and codeine in poppy seed extracts and in hydrolysates of serum and urine were confirmed by GC-MS. Therefore a positive finding of morphine or codeine in blood and urine may sometimes be due to ingestion of poppy seeds.

Excretion of codeine and morphine following ingestion of poppy seeds.

After the ingestion of three poppy-seed bagels, the following codeine and morphine concentrations were determined in the urine: 214 ng/mL codeine and 2797 ng/mL morphine at 3 h, and 16 ng/mL codeine and 676 ng/mL morphine at 22 h. This work indicates that a positive finding of codeine or morphine in the urine of an individual does not necessarily indicate heroin, morphine, or codeine use.

The detection and characterization of controlled drugs imported into the United Kingdom.

The Laboratory of the Government Chemist examines most of the drugs that have been seized at the point of entry into the United Kingdom of Great Britain and Northern Ireland and has developed analytical methods for their rapid identification in the field and for more exact analysis and quantitation in the Laboratory. These methods are described for the major types of drugs encountered. Many seizures are examined in greater detail in order to compare samples and to correlate origin with physical and chemical appearance. Information on the procedures necessary to undertake this aspect of work is also presented.

Dopamine biosynthesis at different stages of plant development in Papaver somniferum.

Latex and cell-free extracts of various organs and stages of plant and capsule development in Papaver somniferum, the opium poppy, synthesized dopamine, an alkaloid precursor, from 14C-dopa. The 1000 g X 30 min supernatant from latex of the pedicel-capsule junction converted more dopa than latex supernatant from the upper capsule or lower pedicel regions, although there was more protein in the latex from the capsule. Percent conversion of pedicel-capsule latex into dopamine was maximum in unopened flower buds and decreased within 14 days after flowering. Dopamine biosynthesis in latex and cell-free extracts also varied with the stage of organ development. Extracts from capsule tissue converted more labeled dopa into dopamine than did extracts from pedicels, leaves from vegetative plants at the rosette stage, leaves from flowering plants, or pedicels connected to capsules.

Enzymic conversion of reticuline to salutaridine by cell-free systems from Papaver somniferum.

A cell-free extract from the opium poppy, Papaver somniferum, was prepared that utilized hydrogen peroxide to convert (+/-)-[3-3H]reticuline to [3H]salutaridine in 80-85% yield based on consumed [3-3H]reticuline. The phenolic-coupling enzyme activity was not detected in the crude homogenate of whole plants. Reticuline conversion to salutaridine was accomplished only after methods were developed to separate both alkaloid and hydrogen peroxide degrading activities from the desired enzyme activity by centrifugal fractionation of carefully prepared stem and root extracts. Purity of the enzymatically produced [3H]salutaridine was established by chromatographic methods and synthetic conversion to [3H]-thebaine.