SUKRASNO SITI KUSMARDIYANI

Transcription

1 SUKRASNO SITI KUSMARDIYANI

2 SCOPE OF MEASUREMENT Biological Physical Chemical Physicochemical

3 BIOLOGICAL PARAMETERS TO BE MEASURED

4 Biological Parameters Crude Drug Chemical Constituents Quality Biological Activity Assay

5 Biological Activity Cytotoxic Brine shrimp lethality test Toxic Fish index Increase membrane permeability Hemolytic test

6 Artificial sea water (ASW) 38 gram sea salt/l EXTRACTS / FRACTIONS / ISOLATES Brine shrimp eggs Adults have three eyes and 11 pairs of legs and can grow to about 15 millimetres in size. 2 days 10 nauplii ( á ~0.5 mm) Evaporate + ASW 5 ml BIOASSAY BRINE SHRIMP LETHALITY TEST - Explain - 1 day Count the number of dead nauplii, use Program Finney A brine shrimp hatchery made from a plastic soap dish and placed in the light. The shrimp should hatch within 24-48h. Significant if LD 50 <30 ug/ml. LD 50 KUSMARDIYANI

7 Translate and discuss. Indeks ikan adalah bilangan atau angka yang menunjukkan pada pengenceran berapa larutan suatu zat membunuh 3 dari 5 ekor ikan yang panjangnya 2 4 cm dalam waktu satu jam. Species: Leuciscus rutilus Carracius vulgaris Tillapia mossambica (mujair) Ikan seribu SK Tanda-tanda keracunan saponin, diamati setelah disimpan selama 24 jam dalam lemari pendingin: Insang menjadi merah, pembuluh darah insang membengkak. (Mati karena sebab lain tidak menyebabkan insang merah) Apandi, A., 1959, Penentuan kekuatan beberapa bahan tumbuhan dengan cara biologi, Suara Farmasi,

8 HEMOLYSIS Red blood cells without (left and middle) and with (right) hemolysis. Note that the hemolyzed sample is transparent, because there are no cells to scatter light. -Wikipedia-

9 PHYSICAL PARAMETERS TO BE MEASURED

10 PHYSICAL PROPERTIES Crude drug Powder Organoleptic Microscopic Morphology Colour Odour Taste Sound Extract Chemical Component Physical property of the major component

11 Physical properties related to liquid state Density Refractive index Boiling point Optical Rotation Solubility/miscibility

12 Physical properties of the major component Swelling capacity swelling index Decrease surface tension foaming index Pungency Scoville index Bitter bitterness index Sweet sweetness index Solubility solvent extractive Ignitability ash residue

13 CHEMICAL PARAMETERS TO BE MEASURED

14 CHEMICAL PROPERTIES Crude Drug Chemical component Chemical group Reactivity

15 Chemical Groups Alkaloid Flavonoid Tannin Quinone Steroid/Triterpenoid..

16 Nucleic acid Plant identification RFLP (Restriction fragment length polymorphism) DNA Restrictive endonuclease Gel Electrophoresis Finger printing (DNA fragment profile) RAPD (Random amplified polymorphism of DNA) DNA PCR Gel electrophoresis Much more simple DNA fragment profile

17 PHYSICOCHEMICAL PARAMETERS TO BE MEASURED

18 PHYSICOCHEMICAL PROPERTIES Spectrophotometry/Spectrometry Chromatography

19 PHYSICOCHEMICAL PROPERTIES Spectrophotometry UV-Vis IR Spectrometry Mass Spectrometry NMR Proton NMR 13 C-NMR --- get more recent information ---

20 UV-Vis Spectrophotometry Qualitative measurement Quantitative measurement Chlorophyl Carotenoid Piperin Curcuminoid --- get more recent information ---

21 IR Spectrophotometry Not much use in analytical pharmacognosy Suitable for volatile oils IR profile can be used as one of identity parameter. --- get more recent information ---

22 --- get more recent information --- Mass Spectrometry Not much use in analytical pharmacognosy. Its combination with gas chromatography: very useful tool in volatile oil analysis. In addition to finger printing, mass spectrum of each component can be obtained and identified base on data base present in system.

23 --- get more recent information --- NMR Spectrometry Not much use in analytical pharmacognosy. Very important instrument for structure elucidation of pure compound.

24 CHROMATOGRAPHY Paper chromatography Thin layer chromatography Column chromatography HPLC GC --- get more recent information ---

25 Paper Chromatography Suitable for polar compound, such as: carbohydrates, phenolics, flavonoids. Limited variation of solvent can be used. --- get more recent information ---

26 Thin Layer Chromatography Very widely used in analytical pharmacognosy. Finger printing of crude drug. Determination of marker. Chromatographic profiling. Can be eluted vertically or circularly. Various composition of solvent can be used. --- get more recent information ---

27 THIN LAYER CHROMATOGRAPHY OF CINCHONA BARKS --- Explain --- Developing system: chloroform - diethylamine (9:1) Spray reagent: (a) 10% H 2 SO 4 in ethanol UV365nm (b) 10% H 2 SO 4, then iodoplatinate reagent visible light 1 = Cinchona calisaya extract 2 = Cinchona succirubra extract TC = a mixture of cinchona alkaloids T1 = quinine T2 = sinchonidine T3 = quinidine T4 = sinchonine (a) (b) Wagner, TLC Atlas..

28 ATLAS OF PLANT DRUG ANALYSIS RUTAE HERBA p144 Wagner, TLC Atlas.. BELADONNAE, HYOSCYAMI, STRAMONII FOLIUM, p49

29 Column Chromatography Suitable for sample preparation. Mostly use to fractionate chemical constituents. Example: Analysis of pesticide residue. Sample clearance prior to HPLC analysis. --- get more recent information ---

30 HPLC High performance liquid chromatography. Very widely used in analytical pharmacognosy. Can be used for qualitative or quantitative analysis. Detection system varies. Coupled with Diode Array Detector provide more information on identity of the peak. --- get more recent information ---

31 HPLC hyphenated Combined with MS (LC-MS) provide better performance of the device. More versatile. Higher sensitivity for quantitative analysis. Better information on identity of each peak by the availability of the mass spectrum of each peak. --- get more recent information ---

32 Gas Chromatography GC is very suitable for analysis of volatile oil. It is also used for the analysis of pesticide residue. Example: Analysis of Peppermint Oil

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35 SCOPE OF MEASUREMENT Biological Physical Chemical Physicochemical Find one example of the monographs, read carefully, and discuss the scope of measurement.

36 Farmakope Indonesia Edisi IV p an example --

37 Farmakope Indonesia Edisi IV p226.

38 Find: specific and non specific parameters for this example of crude drug. Discuss.

39 Find: specific and non specific parameters for this example of crude drug. Discuss.

40 Find: specific and non specific parameters for this example of crude drug. Discuss.

41 Find: specific and non specific parameters for this example of crude drug. Discuss.

42 I. Pharmacopoeial summaries for quality assurance purposes.. Botanical features. Identity tests. Purity requirements. Chemical assays. Major chemical constituents II. Review of scientific research. Clinical applications of plant material. Detailed pharmacological information :. contraindications. warnings. precautions. adverse reactions. dosage

43 VOLUME 1 v v v v v v v v Bulbus Allii Cepae Bulbus Allii Sativi Aloe Aloe Vera Gel Radix Astragali Fructus Bruceae Radix Bupleuri Herba Centellae Flos Chamomillae Cortex Cinnamomi Rhizoma Coptidis Rhizoma Curcumae Longae Radix Echinaceae Herba Echinaceae Purpureae Herba Ephedrae v v v v v v v v v Folium Ginkgo Radix Ginseng Radix Glycyrrhizae Radix Paeoniae Semen Plantaginis Radix Platycodi Radix Rauwolfiae Rhizoma Rhei Folium Sennae Fructus Sennae Herba Thymi Radix Valerianae Rhizoma Zingiberis

44 VOLUME 3 v v v v Fructus Ammi Majoris Fructus Ammi Visnagae Fructus Anethi Aetheroleum Anisi Fructus Anisi Semen Armenicae Flos Arnicae Folium Azadirachti Oleum Azadirachti Flos Carthami Stigma Croci Fructus Foeniculi Radix Gentianae Luteae Radix Gentianae Scabrae Gummi Gugguli Radix Harpagophyti v Rhizoma Hydrastis Radix Ipecacuanhae Aetheroleum Lavandulae Flos Lavandulae Strobilus Lupuli Gummi Myrrha Herba Passifl orae Testa Plantiginis Radix Rehmanniae Fructus Schisandrae Radix Scutellariae Radix cum Herba Taraxaci Semen Trigonellae Foenugraeci Cortex Uncariae Fructus Zizyphi