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    121

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    Santu Das

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    NATURAL DRUGS THE DEVELOPMENT AND VALIDATION OF THE HPLC METHOD FOR MORPHINE CONTENT DETERMINATION IN POPPY STRAW MAEGORZATA ANYZEWSKA, ELZBIETA WOSTASIK, and IRENA ARENT Department of Plant Materials and Herbal Medicine, Drug Institute, 30/34 Chelmska, Str. 00-725 Warsaw, Poland Absteac:The HPLC wth for morphine coment derma im poppy’ saw has heen developed ad ‘ated Te meth! validation involved repay un Ye following preter asurayslectty isp linearity and its of dteton and determination, neterences a Feaovey. The meth was ted to determine the morphine content plan resores clase by the Law Nevt anne” ava inoxcating fom 0p oN, de ont range of 02 190.27 The determination was Keywords Poppy stan, Papaver somnieran. morphine determination ‘The natural source of morphine is Papaverum somniferum L. of Papaveraceae family. It is a one-year plant reaching the 100-150 cm height, IC blossoms at the end of May and begin. ning of June. The formation of poppy heads and their growing lasts about 10-16 days. The techno: logical maturity to crop seeds is achieved 15 days later when stems and leaves dry out and poppy heads become brownish. The crop made at that ‘moment allows to obtain poppy seeds and heads of highest mozphine content. Poppy straw ~ Fructus Papaveris sine Semine (syn. Capita Papaveris) consists of poppy heads without seeds and pieces of stem of about 7 em in length. Poppy straw contains about 0.2 up to 1% of alkaloids, depending on the type of a plant and atmospheric conditions, which are chemically simi- lar to those present in opium, a concentrated milky liquid obtained by incision of green poppy heads. ‘The most active and important alkaloids are de- rivatives of phenanthrene: morphine, codeine thebaine and derivatives of isochinoline: papave. rine and narcotine (1,2). Morphine is formed as ‘result of a biochemical transformation of tyro- sine through norlaudanosine, reticuline and with the loss of one nitrogen atom. Intermediates of such transformations are thebaine and codeine. The production of morphine on industrial scale began in the 1930s based on the method develo- ped by Janos Kobay (3). Morphine is a narcotic analgesic. It paralyzes the respiratory system and causes spasms. of non-striated muscles, It is used in drugs as mor~ pphine hydrochloride (Morphinum hydrochloricum) and sulfate (Morphinum sulphuricum) when other non-narcotic analgesics are inefficient. Its long lasting administration leads to physical and psychic dependance which is called morphinism (1,4). Due to wide spreading problems of drug addiction in Poland a new Law was introduced on April 24,1997 on Neutralization of drug abuse” (5) according to which the trade of poppy straw wi thout permission is a subject to sentence in prison when a morphine content extends 0.05% Poppy straw and morphine (obtained from it) was classified by the Law on Neutralization of drug abuse” as an intoxicating agent from group I-N. Therefore, methods used for morphine deter- ‘mination should be highly accurate and reproucib- le. The aim of this work was the development of an analytical method able to fulfill those requirements. ‘The plant resources involved 80 samples origina- ting from illegal plantations spread in the whole ‘country of Poland, The subject of our study was poppy straw consisting of the poppy heads without seeds and the pieces of stem of about 7 em in Tength (according to the definition of the Law). ‘Analytical methods present in the literature used to determine morphine, involve standard sub- stances, biological materials or different pharma ‘ceutical forms, i. tablets or injections. The deve- lopment of the method used for quantitative evalu- ation of plant resources had to take into account their complex character and variable chemical co- position, The selection of HPLC method was justified due to its growing application among other 122 MAEGORZATA ANYZEWSKA ct al methods such as GC (6), TLC coupled with den sitometry (6), electrophoresis and others, used in routine analysis involving separation of compo- rents in complex mixtures, Moreover, the results of ‘morphine determination in poppy straw were com- pared with TLC-densitometric results for evalua tion purposes. The method and results for den- sitometrc determinations will be presented in a fo- rthcoming paper. In order to find the most effective chromato- ‘graphic conditions, the reverse phase approach ‘was applied. The results were obtained on several silica based chromatographic columns, such as: RP-C8, RP-18, RP-CN and RP-C18 AB, Accor-

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