E tg/166/4(proj. 3) Original
|
Спампаваць 476.87 Kb.
|
8.Explanations on the Table of Characteristics8.1 Explanations covering several characteristicsCharacteristics containing the following key in the second column of the Table of Characteristics should be examined as indicated below: (a) 10-12 true leaves stage (prior to internode elongation); (b) The pedicel is in hook stage; (c) Time of flowering (when the first flower opens on the main stem at 10 % of the plants); (d) Full blossom; (e) 10-14 days after the petals drop down on main stem; (f) Mature, dry capsule of main stem. 8.2 Explanations for individual characteristicsAd. 2: Rosette leaf: white spots Ad. 3: Rosette leaf: color Ad. 4: Rosette leaf: waxiness The observations of white spots, color and waxiness should be made on the upper side of the rosette leaf. Ad. 5: Rosette leaf: depth of lobes
Ad. 6: Main stem: length 
Ad. 7: Stem: anthocyanin coloration Ad. 8: Stem: hairiness The observation of anthocyanin coloration and hairiness should be made between the capsule and the upper stem leaf. Ad. 9: Flower bud: anthocyanin coloration
Ad. 11: Petal: blotch
Ad. 13: Petal: extension of blotch The measurement should be made at the widest point of petal.
Ad. 14: Petal: incisions
Ad. 18: Capsule: shape of longitudinal section
Ad. 19: Capsule: shape of base Ad. 20: Capsule: length
Ad. 22: Capsule: ribbing For the observation of ribbing, the capsule should be touched. Ad. 23: Capsule: dehiscence For the observation of dehiscence the capsule should be held upside-down and shaken. If seeds do not fall out, the capsule is indehiscent (1). If seeds fall out, the capsule is dehiscent (2). Ad. 24: Stigmatic disc: shape 
Ad. 26: Stigmatic disc: apex of lobe 
Ads. 29-32: Capsule: determination of alkaloid contents: morphine, codeine, thebaine and narcotine Determination of Morphine, Codeine, Thebaine, Papaverine and Narcotine content in poppy capsule HPLC method, MS detection 1. Scope Determination of Morphine, Codeine, Thebaine, Papaverine and Noscapine content in poppy capsule for qualification purposes. Limit Of Detection (LOD): 10 mg/kg/component Limit Of Quantitation (LOQ): 50 mg/kg/component 2. Principle The sample is extracted with methanol containing 1 ml of cc. hydrochloric acid/litre. The alkaloid content of the extract is determined by HPLC-MS system using RP C18 column. External standards are used for qualitative and quantitative determination. 3. Procedure 3.1. Sample preparation The receipt sample is weighted and dried to air-dry condition. The stalk, the crop and the seeds are separated. The stalk is ground using 0,5 mm sieve. 3.2. Extraction and clean-up Weigh 0,2 g of grinded sample and add 100 ml methanol-HCl solution (1 ml cc. HCl/litre methanol. Keep in ultrasonic bath 30 minutes. Filter and this solution inject to the HPLC column. 3.3. HPLC measurement The determination of the alkaloid content is performed by MS detection (SIM mode) after separation using reversed phase C18 column. HPLC conditions The HPLC conditions are advised listed below, but any other conditions can be used if those give suitable results. Chromatographic column: NUCLEODUR C-18 Gravity 150*4.6mm*5μm or equivalents. Mobile phase A eluent: HPLC grade methanol B eluent: 2 g Ammonium-acetate/litre HPLC grade water Gradient: 0-4 min. 70% B 4-14 min. 10% B-ig linear gradient 14-20 min. 10% B Post time: 5 min.
0.9 cm3/min. Detector MS SIM APCI: 2-20 perc: 286.0 AMU Positive 300.0 AMU Positive 312.0 AMU Positive 340.0 AMU Positive 414.0 AMU Positive Injected volume: 2 l For qualitative and quantitative determination used analytical grade standard solutions in HCL methanol (1 ml cc. HCl/litre methanol) solvent. Calibrate according to ESTD method. 4. Expression of the results The results are expressed in mg/kg referred to air-dry material. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
