Codeine technique

Biosynthetic research relating to the isoquinoline family was extremely successful, with such important members as morphine [3, 14], codeine [3, 15] or berberine [3, 14,16-18]. Extensive efforts have provided details pertaining to multiple sets of enzymes participating in the biosynthesis of the alkaloids above, in many cases with the help of plant cell suspension culture techniques. Since 1988, when the breakthrough in cloning of cDNA from alkaloid biosynthesis occurred [19, 20], a significant number of enzymes known from the indoles and isoquinolines biosynthesis have been isolated, their biochemical properties described and the majority of their corresponding cDNAs cloned and functionally over-expressed in non-plant hosts such as Escherichia coli, yeast or insect cells. 

Codeine is a component of gum opium and can also be produced by methylation of morphine using known prior art techniques. 

Alkaloid biosynthetic pathways are under strict regulation in plants. Until now, our limited knowledge of the fundamental mechanisms involved in the control of alkaloid metabolism has severely restricted our ability to harness the vast biotechnological potential of these important secondary pathways. For example, the use of plant cell cultures for the commercial production of pharmaceutical alkaloids has not become a reality despite decades of empirical research. The application of traditional and modem biochemical, molecular, and cellular techniques has revealed important clues about the reasons why C. rosens cultures accumulate tabersonine and catharanthine, but not vindoline or vinblastine, and why opium poppy cultures produce sanguinarine, but not codeine or morphine. The inability of dedifferentiated cells to accumulate certain metabolites was interpreted as evidence that the operation of many alkaloid pathways is tightly coupled to the development of specific tissues. Recent studies have shown that alkaloid pathways are regulated at multiple levels,... 

Enzymatic digestion gives equally excellent results for the four opiates. Comparing this method with SFE shows that the two techniques are comparable for codeine and ethylmorphine, but show differences when it comes to 6-MAM and morphine. In fact the higher extraction yield for morphine and the lower yield for... 

Codeine occurs in opium in relatively small amounts (—0.3%) and yet it is in great demand as an analgesic. Much of the morphine derived from opium is therefore converted by methylation to codeine. Conventional methylation techniques have been employed, including dimethyl sulfate, diazomethane, and phenyltrimethylammonium chloride. 

The most developed areas for imprinted polymers are for sequestration and separation. Several reported imprinted polymer sensors have in actuality been pretreatments by separation to allow a selective determination using a general detection method. The adaptation of this to an FIA system would be relatively simple. For example, Kriz et al. [31] report a sensor for morphine (see Chapter 18). The method of morphine detection involved two steps. The first step was to immobilise the morphine by loading it on the imprinted polymer. In the detection step, the morphine was released from the column by elution of an electro-inactive competitor (codeine) and the released morphine was detected by an amperometric method. The polymer was tested after exposure to extremes of heat and chemicals and proved resilient. This method would probably be suitable for automation as a flow injection technique. 

Opium poppy Papaver somniferum L., Papaveraceae) is one of the most important medicinal plants and has been cultivated since early centuries. Opium, the dried cytoplasm of a specialized internal secretory system called the laticifer, is normally collected from the unripe capsule. It is the source for the commercial production of medicinally important alkaloids, morphine, codeine, thebaine, noscapine and papaverine [130, 131], Fig. (61). Morphine, which has strong addictive property, is still the most effective analgesic for the treatment of mortal cancer patients in modem medicine. Codeine is commonly used as an antitussive. However, field cultivation of this plant has been limited since 1953 by the United Nations Opium Conference Protocol to prevent narcotic crimes. Therefore, establishing tissue culture technique for the production of morphinan alkaloids seems to be desirable not only for medicinal purpose but also for decreasing abuse of opiates. 

In the following years cytochrome P450 was also found as an integral part of the 1 l 3-hydroxylase system in adrenal mitochondria Its involvement in the liver microsomal monooxygenation of codeine, acetanilide and cyclohexane was established using the technique of the photochemical action spectrum (Fig. 2). 

One recent development that offers scope for direct injection of aqueous samples into an MS system is the high-field, asymmetric waveform ion mobility spectrometer (FAIMS). By interfacing the FAIMS technology to an ESI—MS instrument a significant reduction in detection limits has been achieved for molecules of <300 Da. Preconcentration is unnecessary and analytical times are significantly shortened. Recent appHcations of this technique include naphthenic and haloacetic acids in water and amphetamines, morphine and codeine... 

In an extremely powerful technique, Dams et al. [540] separated 18 opiates and their derivatives (morphine, codeine, naloxone, hydrocodone, papaverine, dextromethorphan, noscapine, bupremorphine, methadone, heroin, thebacone, ethylmor-phine, 6-monoacetylmorphine, acetyldihydrocodeine, acetylcodeine, normethadone, normorphine, norcodeine) with baseline resolution in <12 min This was accomplished on a fast LC phenyl column 53 x 7 mm (2 = 280 nm) using a simple 100/0—>50/50 (at 10min)—> 0/100 (at 12min hold Imin) (90/5/5 water/ methanol/acetonitrile with 50 mM ammonium acetate)/(50/50 methanol/ acetonitrile with 50 mM ammonium acetate) gradient. The mobile phase components were adjusted to pH pp 4.5 with formic acid prior to use. The phenyl column was chosen ova- the conventional Cg or C g columns because of the increased selective interaction with analytes containing the planar phenyl functional group. Detection limits from 50 to 450ng/mL (analyte dependent) were reported. 

GLC/TLC Analysis of Codeine and Morphine in Urine Via Derivatization Techniques... 

Rapid Gas-Liquid Chromatographic Procedure for the Determination of Codeine and Norcodeine in Biological Fluids Based on Micro-phase Extraction Techniques J. Pharm. Pharmacol. 27(12) 937-939... 

One such technique, called lead modification, enables scientists to identify the portion of a compound responsible for its medicinal properties and then to design similar compounds with better properties. We will see an example of this technique, specifically, where the discovery of morphine led to the development of a whole family of potent analgesics (codeine, heroin, methadone, and many others). 

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