scope of measurement
TRANSCRIPT
SUKRASNO SITI KUSMARDIYANI
SCOPE OF MEASUREMENT
Biological Physical Chemical Physicochemical
BIOLOGICAL PARAMETERS
TO BE MEASURED
Crude Drug
Chemical Constituents
Biological Activity
Quality
Assay
Biological Parameters
Toxic
Cytotoxic
Increase membrane
permeability
Brine shrimp lethality test
Fish index
Hemolytic test
Biological Activity
EXTRACTS / FRACTIONS / ISOLATES
2 days
1 day
LD50
10 nauplii( á ~0.5 mm)
Evaporate + ASW 5 mL
Artificial sea water (ASW) 38 gram sea salt/L
Brine shrimp eggs
Count the number of dead nauplii, use Program Finney
BRINE SHRIMP LETHALITY TEST
A brine shrimp hatchery made from a plastic soap dish and placed in the light. The shrimp should hatch within 24-48h. Significant if LD50 <30 ug/mL. KUSMARDIYANI
BIOASSAY- Explain -
Adults have three eyes and 11 pairs of legs and can grow to about 15 millimetres in size.
Indeks ikan adalah bilangan atau angka yang menunjukkan pada pengenceranberapa larutan suatu zat membunuh 3 dari 5 ekor ikan yang panjangnya 2 – 4 cmdalam waktu satu jam.
Species:
Leuciscus rutilusCarracius vulgarisTillapia mossambica (mujair) Ikan seribu
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, 23 – 28.
Translate and discuss.
© SK
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.
HEMOLYSIS
-Wikipedia-
PHYSICAL PARAMETERS
TO BE MEASURED
PHYSICAL PROPERTIES
Crude drug
Extract
Organoleptic
Microscopic
Chemical Component
Powder
MorphologyColourOdourTaste Sound
Physical property of the major component
Density Refractive index Boiling point Optical Rotation Solubility/miscibility
Physical properties related to liquid state
Swelling capacity swelling index Decrease surface tension foaming index Pungency Scoville index Bitter bitterness index Sweet sweetness index Solubility solvent extractive Ignitability ash residue
Physical properties of the major component
CHEMICAL PARAMETERS
TO BE MEASURED
Crude Drug
Chemical component
ReactivityChemical group
CHEMICAL PROPERTIES
Chemical Groups
Alkaloid Flavonoid Tannin Quinone Steroid/Triterpenoid ……. …….
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
PHYSICOCHEMICAL PARAMETERS
TO BE MEASURED
• Spectrophotometry/Spectrometry
• Chromatography
PHYSICOCHEMICAL PROPERTIES
• Spectrophotometry–UV-Vis– IR
• Spectrometry– Mass Spectrometry– NMR• Proton NMR• 13C-NMR
PHYSICOCHEMICAL PROPERTIES
--- get more recent information ---
UV-Vis Spectrophotometry
– Qualitative measurement– Quantitative measurement
• Chlorophyl• Carotenoid• Piperin• Curcuminoid
--- get more recent information ---
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 ---
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.
--- get more recent information ---
NMR Spectrometry
• Not much use in analytical pharmacognosy.
• Very important instrument for structure elucidation of pure compound.
--- get more recent information ---
CHROMATOGRAPHY
• Paper chromatography• Thin layer chromatography• Column chromatography• HPLC• GC
--- get more recent information ---
Paper Chromatography
• Suitable for polar compound, such as: carbohydrates, phenolics, flavonoids.
• Limited variation of solvent can be used.
--- get more recent information ---
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 ---
THIN LAYER CHROMATOGRAPHY OF CINCHONA BARKS
Developing system: chloroform - diethylamine (9:1) Spray reagent: (a) 10% H2SO4 in ethanol UV365nm
(b) 10% H2SO4, 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..
--- Explain ---
RUTAE HERBA p144
BELADONNAE, HYOSCYAMI, STRAMONII FOLIUM, p49
ATLAS OF PLANT DRUG ANALYSIS
Wagner, TLC Atlas..
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 ---
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 ---
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 ---
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
-------
SCOPE OF MEASUREMENT
Biological Physical Chemical Physicochemical
Find one example of the monographs, read carefully, and discuss the scope of measurement.
Farmakope Indonesia Edisi IVp225. -- an example --
Farmakope Indonesia Edisi IV
p226.
Find: specific and non–specific parameters for this example of crude drug.
Discuss.
Find: specific and non–specific parameters for this example of crude drug. Discuss.
Find: specific and non–specific parameters for this example of crude drug. Discuss.
Find: specific and non–specific parameters for this example of crude drug. Discuss.
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
Bulbus Allii CepaeBulbus Allii SativiAloeAloe Vera GelRadix AstragaliFructus BruceaeRadix BupleuriHerba CentellaeFlos ChamomillaeCortex CinnamomiRhizoma CoptidisRhizoma Curcumae LongaeRadix EchinaceaeHerba Echinaceae PurpureaeHerba Ephedrae
Folium GinkgoRadix GinsengRadix GlycyrrhizaeRadix PaeoniaeSemen PlantaginisRadix PlatycodiRadix RauwolfiaeRhizoma RheiFolium SennaeFructus SennaeHerba ThymiRadix ValerianaeRhizoma Zingiberis
--------------------- VOLUME 1
vvvv
v
v
v
v
v
v
vvvvvvv
Fructus Ammi MajorisFructus Ammi VisnagaeFructus AnethiAetheroleum AnisiFructus AnisiSemen ArmenicaeFlos ArnicaeFolium AzadirachtiOleum AzadirachtiFlos CarthamiStigma CrociFructus FoeniculiRadix Gentianae LuteaeRadix Gentianae ScabraeGummi GugguliRadix Harpagophyti
--------------------- VOLUME 3
Rhizoma HydrastisRadix IpecacuanhaeAetheroleum LavandulaeFlos LavandulaeStrobilus LupuliGummi MyrrhaHerba Passifl oraeTesta PlantiginisRadix RehmanniaeFructus SchisandraeRadix ScutellariaeRadix cum Herba TaraxaciSemen Trigonellae FoenugraeciCortex UncariaeFructus Zizyphi
v
vv
v
v