INTEGRACIN DE LA ACTIVIDAD FARMACUTICA PARA LOGRAR UN RESULTADO PRCTICO SOCIAL, COMERCIAL Y/o ECONMICO EN LA INVESTIGACIN COMO ACTIVIDAD PROFESIONAL

For several thousand years, man has used herbs and potions as medicines, but it is only since the mid nineteenth century that serious efforts were made to isolate and purify the active principles of these remedies. Since then, a large variety of biologically active compounds have been obtained and their structures determined.

DRUGS DEVELOPMENT

Ancient man learned from trials and error, which plants were useful or not.

 These natural products became the lead compounds for a major synthetic effort where chemists made literally thousands of analogues in an attempt to improve of what nature had provided. The vast majority of this work was carried out with no real design or reason, but out of the results came an appreciation of certain tactics which generally worked.

DRUGS DEVELOPMENT

Dioscorides, a Greek pharmacobotanist who lived in the first century A.D., wrote his De Materia Medica concerning medical matter in 78 A.D. in which he described about 600 plants that were known to have medicinal properties. Of these, a surprising large number are still important in modern medicine. Aloe, belladonna, colchicum, ergot, hyoscyamus, and opium are a few that were used then in much the same manner as they are used today.

Pedanio (o Pedacio) Dioscrides Anazarbeo

Galen (131-200 A.D.) was a Greek pharmacistphysician who lived in Rome and who described the method of preparing formulas containing plant and animal drugs. He devoted considerable time to compiling this knowledge, which was distributed throughout 20 books. As a tribute to his accuracy in recording his observations, the term galenical pharmacy was originated.

Galeno de Prgamo

Galen practiced and taught both Pharmacy and Medicine in Rome; his principles of preparing and compounding medicines ruled in the Western world for 1,500 years; and his name still is associated with that class of pharmaceuticals compounded by mechanical means - galenicals. He was the originator of the formula for a cold cream, essentially similar to that known today. Many procedures Galen originated have their counterparts in today's modern compounding laboratories.

 Screening of natural sources for biological activity. Etnopharmacolgy information. Standardization. Extraction, isolation and purification of the active principle. Determination of structure. Structure activity relationships Preclinical and clinical evaluation. Formulation. Design and synthesis of novel drug structure and analogues.

DEVELOPMENT OF A NOVEL DRUG

COMPOUND FROM NATURAL SOURCES PLAY FOUR SIGNIFICANT ROLES IN MODERN MEDICINE. 1-THEY PROVIDE DRUGS IMPOSSIBLE TO PRODUCE COMMERCIALLY BY SYNTHESIS.

2-It supply basic compounds that may be modified slightly to render more effective or less toxic.

3-They are prototypes or models for synthetic drugs possessing physiologic activities similar to the originals.

EXAMPLES OF THERAPEUTICALLY ACTIVE STEROIDS

4- They supply structures without activity but with chemical modification could be potent drugs
SEX HORMONES

CORTICOIDS

ORAL CONTRACEPTIVES

DIURETIC STEROIDS

Quality control have to assure:

The correct botanical identity of the drug The purity of the material used Avoid the presence of insects, mites, bacteria, fungi, heavy metals, etc. That the required level of active compounds or a defined level of biological activity is reached.

Quality control is a multistep process that covers all stages from the growing of the plant until the final control of the commercial product and the evaluation of its stability and quality all over the time.

Standardization The concept of standardization is relatively new for phytomedicine, but it is rapidly becoming essential to ensure that patients are provided with high-quality botanical products.

STANDARDIZATION
Why is standardization necessary and important?

Reproducible higher quality of the product. Provided that the product compared with standards could be included in Formularies. Standardization allows comparison of the clinical effectiveness, pharmacological effects and side effects of a series of vegetable drugs(e.g. against placebo). Such products give patients greater (objective and subjective) security and thus increase the amount of people using herbal products. It is a key task, which can be performed by a pharmacist and is in fact done by pharmacists in Switzerland, Germany ,France and others.

Precision of measurement Quantities and volumes The quantities and volumes of the materials and reagents used in the tests must be measured with adequate precision, which is indicated in the following way: A value of: 20.0 means not less than 19.5 and not more than 20.5 2.0 means not less than 1.95 and not more than 2.05 0.20 means not less than 0.195 and not more than 0.205. Temperature Temperature measurement is indicated in a manner similar to that given for quantities and volumes. Storage conditions given in general terms refer to the following equivalent temperatures: In a refrigerator 0-6 C Cold or cool 6-15 C Room temperature 15-25 C, or up to 30C depending on climatic zones.

Solubility Unless otherwise specified in the test procedure for the plant material concerned, the approximate solubility of medicinal plant materials should be determined at 20C. Solubility is expressed in terms of "parts", representing the number of milliliters (ml) of the solvent, in which 1 g of the solid is soluble. Descriptive terms are sometimes used to indicate the solubility of a substance, with the following meanings: very soluble less than 1 part freely soluble 1-10 parts soluble 10-30 parts sparingly soluble 30-100 parts slightly soluble 100-1000 parts very slightly soluble 1000-10 000 parts practically insoluble more than 10000 parts

Containers The container and its closure must not interact physically or chemically with the material within in any way that would alter its quality. Size of cut Medicinal plant materials are used either whole, or in cut or powdered form. Cut medicinal plant materials are prepared by cutting or crushing the plant into small pieces. The cut is graded according to the aperture size of the mesh of the sieve through which the material will pass, and is indicated as follows: Aperture size (mm) coarse cut 4.00 medium cut 2.80 fine cut 2.00

Sampling of material - organoleptic characteristics (colour, texture and odour); - presentation of the material (raw, cut, crushed, compressed); - the presence of admixtures, foreign matter (sand, glass particles, dirt), mould, or signs of decay; - the presence of insects; - the presence of packaging material originating from poor or degraded containers.

Foreign matter is material consisting of any or all of the following: - parts of the medicinal plant material or materials other than those named with the limits specified for the plant material concerned; - any organism, part or product of an organism, other than that named in the specification and description of the plant material concerned; - mineral admixtures not adhering to the medicinal plant materials, such as soil, stones, sand, and dust.

Macroscopic and microscopic examination

Medicinal plant materials are categorized according to sensory, macroscopic and microscopic characteristics. An examination to determine these characteristics is the first step towards establishing the identity and the degree of purity of such materials, and should be carried out before any further tests are undertaken. Wherever possible, authentic specimens of the material in question and samples of pharmacopoeia quality should be available to serve as a reference.

Determination of ash
The ash remaining following ignition of medicinal plant materials is determined by three different methods which measure total ash, acid-insoluble ash and water-soluble ash. The total ash method is designed to measure the total amount of material remaining after ignition. This includes both "physiological ash", which is derived from the plant tissue itself, and "non-physiological" ash, which is the residue of the extraneous matter (e.g. sand and soil) adhering to the plant surface. Acid-insoluble ash is the residue obtained after boiling the total ash with dilute hydrochloric acid, and igniting the remaining insoluble matter. This measures the amount of silica present, especially as sand and siliceous earth. Water-soluble ash is the difference in weight between the total ash and the residue after treatment of the total ash with water.

Determination of extractable matter

This method determines the amount of active constituents extracted with solvents from a given amount of medicinal plant material. It is employed for materials for which as yet no suitable chemical or biological assay exists.

Determination of remaining water

An excess of water in medicinal plant materials will encourage microbial growth, the presence of fungi or insects, and deterioration following hydrolysis. Limits for water content should therefore be set for every given plant material. This is especially important for materials that absorb moisture easily or deteriorate quickly in the presence of water.

Determination of volatile oils

Volatile oils are characterized by their odor, oil-like appearance and ability to volatilize at room temperature. Chemically, they are usually composed of mixtures of, for example, mono-terpenoids, sesqui-terpenoids and their oxygenated derivatives. Aromatic compounds predominate in certain volatile oils. Because they are considered to be the "essence" of the plant material, and are often biologically active, they are also known as "essential oils". The term "volatile oil" is preferred because it is more specific and describes the physical properties.

Determination of bitterness value

Medicinal plant materials that have a strong bitter taste ("bitters") are employed therapeutically, mostly as appetizing agents. Their bitterness stimulates secretions in the gastrointestinal tract, especially of gastric juice.

Determination of haemolytic activity

Many medicinal plant materials, especially those derived from the families Caryophyllaceae, Araliaceae, Sapindaceae, Primulaceae, and Dioscoreaceae contain saponins. The most characteristic property of saponins is their ability to cause hemolysis: when added to a suspension of blood, saponins produce changes in erythrocyte membranes, causing haemoglobin to diffuse into the surrounding medium.

Determination of tannins
Tannins (or tanning substances) are substances capable of turning animal hides into leather by binding proteins to form water-insoluble substances that are resistant to proteolytic enzymes. This process, when applied to living tissue, is known as an "astringent" action and is the reason for the therapeutic application of tannins. Chemically, tannins are complex substances; they usually occur as mixtures of polyphenols that are difficult to separate and crystallize. They are easily oxidized and polymerized in solution; if this happens they lose much of their astringent effect and are therefore of little therapeutic value.

Determination of swelling index

Many medicinal plant materials are of specific therapeutic or pharmaceutical utility because of their swelling properties, especially gums and those containing an appreciable amount of mucilage, pectin or hemicellulose.

Determination of pesticide residues

Medicinal plant materials are liable to contain pesticide residues which accumulate from agricultural practices, such as spraying, treatment of soils during cultivation, and administration of fumigants during storage. It is therefore recommended that every country producing medicinal plant materials (naturally grown or cultivated) should have at least one control laboratory capable of performing the determination of pesticides

Determination of arsenic and heavy metals

Contamination of medicinal plant materials with arsenic and heavy metals can be attributed to many causes including environmental pollution and traces of pesticides. Determination of microorganisms Medicinal plant materials normally carry a great number of bacteria and moulds, often originating in soil. While a large range of bacteria and fungi form the naturally occurring microflora of herbs, aerobic spore-forming bacteria frequently predominate. Current practices of harvesting, handling and production may cause additional contamination and microbial growth. The determination of Escherichia coli and moulds may indicate the quality of production and harvesting practices. Methods for decontamination are restricted. For example, the use of ethylene oxide has been forbidden within countries of the European Union. Treatment with ionizing irradiation is also forbidden or requires a special registration procedure in some countries. In addition, the presence of aflatoxins in plant material can be hazardous to health if absorbed even in very small amounts.

METODOS DE EXTRACCION

CONCEPTO
LA EXTRACCIN SEPARA LAS PORCIONES MEDICINALMENTE ACTIVAS A PARTIR DE LOS TEJIDOS DE PLANTAS Y ANIMALES DE LOS COMPONENTES INERTES DE LOS MISMOS CON DISOLVENTES SELECTIVOS DENOMINADOS MENSTRUOS LOS EXTRACTOS DE PLANTAS MEDICINALES SE DENOMINAN GALNICOS EN HONOR A GALENO, MDICO GRIEGO QUIEN DIO GRANDES APORTES A LA MEDICINA Y LA FARMACIA EN SU EPOCA.

INFUSIN Y DECOCCION
INFUSIN: COLOCANDO LA DROGA 15 MINUTOS EN AGUA FRIA O CALIENTE PROPORCIN 1/10 CUANDO NO HAY SUSTANCIAS DRSTICAS, 1/30 EL GENERAL Y 1/400 CUANDO HAY SUSTANCIAS TXICAS.

MACERACION
SE REMOJA LA DROGA CON EL DISOLVENTE A TEMPERATURA AMBIENTE ENTRE 2-14 DIAS Y MNIMO 7 SI NO SE CONOCE EL TIEMPO DEL PROCESO. SE FAVORECE POR LA AGITACION Y LIGERO CALENTAMIENTO NO EXTRAE TODA LA CANTIDAD DISPONIBLE DE PRINCIPIOS ACTIVOS

Continuous extraction: The plant material is treated with solvents of increasing polarity that dissolve one or some of its components. This methods require specially designed equipments such as the Soxhlet apparatus and the liquid-liquid extractor. The biomass is placed on a Soxhlet thimble constructed of filter paper, through which solvent is continuously refluxed.

LIXIVIACION O PERCOLACION
SE UTILIZA PARA LA PREPARACION DE EXTRACTOS FLUIDOS DONDE EL VOLUMEN FINAL DEL EXTRACTO ES UNO A UNO, VOLUMEN FINAL/ PESO DE DROGA DE PARTIDA. SE REALIZA MEDIANTE UN PERCOLADOR

TIPOS DE EXTRACTOS LIQUIDOS O SEMILIQUIDOS EXTRACTO SLIDO O PILULAR EXTRACTO PULVERIZADO O POLVO SECO

IN THE PHARMACEUTICAL MARKET THERE ARE USUALLY FOUND FOUR TYPES OF EXTRACTS:
Hydro alcoholic extracts
Are the must common and contain the whole diversity of chemical components. The two common different kinds of extracts are:
Fluid extract: concentration of the drug extract until 1 ml contain the extractives of 1 g of plant. Tincture: Concentration of the drug extract to certain percentage. For example: 10% tincture 1 g drug / 10 mL 20% 1 g drug / 5 mL

Dry extracts

Are obtained concentrating the liquid extraction through vacuum evaporation or spray dry system until a powder is obtained. As termolabile substances could change its chemical structure, spray dry is used nowadays. This procedure preserves the enzymatic, vitamin and hormonal content of fresh plant preparations.

Soft extracts Are thick liquids or semisolid masses that are obtained evaporating the extraction solvent without reaching to dryness. In general, each gram of the soft extract is equivalent to 4-6 g of extractives from each 100g of the raw material.

Oily extracts

When the substance of interest is hydrophobic and the evaporation of the extract finally is oily in its consistence

ANALISIS DE CALIDAD Y SEGUIMIENTO DE LA ESTABILIDAD

Descripcin pH ndice de refraccin Peso especfico Slidos solubles Cenizas y contenido de metales Determinaciones cualitativas Anlisis cromatogrfico Contenido de alcohol Anlisis capilar

 Phytochemical investigations.
Preliminary phytochemical screening. Quantitative chemical examination.
Some qualitative tests are performed directly on the fresh plant or dried crude drug to identify specific groups or types of components. The purity of a crude drug is determined by quantitative estimation of active chemical constituents present in them. The method may be useful in determining single active constituents or the group of related constituents present in the same drug.

WHAT IS SYNERGY?
Synergy means working together.

It happens that the combined action of constituents is greater than would be expected from individual constituents.
In Pharmacognosy, synergy has its specific definition. The term is applied to describe any kind of interaction between constituents of a single extract, as well as the components of a mixture of herbs that may potentiate some therapeutic effects.

Whether an effect is truly synergistic, or merely additive, is something that must be established, and evidence has to prove it conclusively.

ADVANTAGES AND DISADVANTAGES OF MIXTURES/COMBINATIONS

In general, synergistic or other interactive effects of a mixture is considered to be positive, when the low dose used demonstrate a benefit in the treatment. Alternative medical systems such as Ayurveda (Indian medicine system) and traditional Chinese medicine consider mixtures as an integral part of the treatment.

TOXICITY OF HERBAL CONSTITUENTS

Most common herbal remedies are fairly safe in the clinical use, not because they are natural, but because the long history of use has uncovered some of the adverse effects. Traditional use is not always a reliable indication of safety and, because many patients do not consider phytomedicine to be drugs. An association may have not be made between the remedy and the problem. Alternatively, a long interval between the dose of the medicine and the cause of a reaction make the connection difficult. As with all medicines, side effects, which are outlined in the monograph for each herb, and interaction with other drugs are possible. But these are consequence of the therapeutic use of the herb and the usual risk : benefit ratio is evident.

FILTRATION. PROVIDES THE EASIEST AND MOST OBVIOUS METHOD OF SAMPLE SEPARATION, NECESSARY FOR COUNTER CURRENT CHROMATOGRAPHY ORE ANY OTHER SEPARATION PROCEDURE. THIS CAN TAKE THE FORM OF THE PASSAGE OF A SAMPLE SOLUTION THROUGH A FILTER PAPER OR SINTERED GLASS FUNNEL, IN ORDER TO REMOVE PARTICULATE AND INSOLUBLE MATERIAL

PRECIPITATION. ANOTHER PRELIMINARY PURIFICATION OF SAMPLES IS BY MEANS OF PRECIPITATION. THIS IS A METHOD A VERY OFTEN EMPLOYED METHOD THAT CONSIST IN POURING A CONCENTRATED SOLUTION OF ONE EXTRACT IN ONE SOLVENT INTO ANOTHER OF DIFFERENT POLARITY. THE PRECIPITATED IS COLLECTED AND THE PRECIPITATION CAN BE REPEATED SEVERAL TIMES.

Probably the simplest separation method is

partitioning,
which is used as an initial extract purification step. Partitioning uses two immiscible solvents to which the extract is added;

this can be sequential by using immiscible organic solvents of increasing polarity.

STRAIGHTFORWARD SOLVENT PARTITION REMOVE A LARGE PORTION OF EXTRANEOUS CONSTITUENTS AND, ESPECIALLY WHEN USED IN CONJUNCTION WITH A BIOASSAY, FRACTIONS ENRICHED IN THE SOUGHT FOR CONSTITUENTS ARE RAPIDLY OBTAINED.

MULTIPLE PARTITION STEPS PROVIDE ANOTHER POSSIBILITY FOR THE PRELIMINARY PURIFICATION OF SAMPLES WHICH ARE TO BE SEPARATED BY ANY CHROMATOGRAPHIC TECHNIQUE. COUNTER CURRENT DISTRIBUTION, GENERALLY IS USEFUL IN THIS RESPECT, ESPECIALLY WHEN SEPARATING LIPOPHILIC FROM MORE POLAR CONSTITUENTS.

PAPER AND THIN LAYER CHROMATOGRAPHY

INTRODUCTION
The paper chromatography and thin layer chromatography (TLC) techniques are similar in that they are both open bed techniques in which substances are separated by the differential migration that occurs when a solvent flows along a thin layer of paper (paper chromatography) or fine powder spread on a glass or plastic plate (TLC).

The solution of components is applied as a spot near one end of a prepared filter-paper strip. The paper is then supported in an airtight chamber which has an atmosphere saturated with the solvent

Line of spot application Solvent line

X X X X X

DEVELOPING AGENTS
After the filter-paper strips or TLC plates have been dried, the positions of the separated components can be revealed by the use of suitable developing agents:
Ninhydrin solution for amino acids Iodine solution (or vapour) or a modified Dragendorffs reagents for alkaloids Ferric chloride solution for phenols Alkali for Anthraquinones derivatives Antimony trichloride in chloroform for steroids and some components of volatile oils Aniline hydrogen phthalate reagent for sugars

TWO-DIMENSIONAL CHROMATOGRAPHY
For the separation of some bioconstituents, it is necessary to use a two-dimensional chromatography using two different solvent systems.
The resolved components of original mixture can be separately eluted from chromatogram by treating the cut-out spots with a suitable solvent and then determined quantitatively by some suitable instrumental methods of analysis, for example, fluorescence analysis, colorimetric, or ultra-violet absorption.

COLUM CHROMATOGRAPHY

CONVENTIONAL OPEN COLUMN CHROMATOGRAPHY IS UNIVERSALLY PRACTICED AS A RESULT OF ITS SIMPLICITY OF OPERATION. AS FAR AS SILICA GEL IS CONCERNED, 30 MG OF SAMPLE LOADING PER G OF SUPPORT ARE FEASIBLE, BUT THIS VERY HIGH CAPACITY IS ONLY POSSIBLE WHEN THE SUBSTANCES TO BE SEPARATE DIFFER GREATLY IN THEIR RF VALUES. LOADING 10 MG OF SAMPLE PER G SUPPORT ARE MORE COMMON. THE LIMITATIONS OF THE METHOD ARE -SLOW SEPARATION -IRREVERSIBLE ABSORPTION OF SOLUTES -INCOMPATIBILITY WITH SMALL GRANULOMETRY PARTICLES

HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)

High performance liquid chromatography (HPLC) is a relatively new comer in the field of liquid chromatography.

HPLC hinges on speed and resolution for its effectiveness. Preparative technique requires another factor named load. Resolution is impaired if the load is too high, column overloading arising from excessive sample mass. The loading capacity depends on variables such as column radius, length, particle diameter and packing density of the support.

Injection valve Filter Column detector Column Filter Eluant reservoir Detector Pump Pressure module

Recorder

Fraction collector

EJEMPLO DE UNA PURIFICACIN SEGUIDA POR HPLC

ESPECTROSCOPIC CHARACTERIZATION
UV
O

BANDA IIO BANDA I 240 - 285 300 - 400

COMPOUND
Flavonols Flavona Flavanons

BAND I
351-385 304-350 sh

BAND II
239-271 239-281 270-295

Flavononols
Isoflavones Chalcons Aurons Anthocyanidins Catequins

sh
sh 340-390 370-430 465-550 sh

245-270
245-270 220-270 220-270 270-280 280

Espectroscopia de resonancia magntica nuclear

La espectroscopa de resonancia magntica nuclear (RMN) es una tcnica empleada principalmente en la elucidacin de estructuras moleculares, aunque tambin se puede emplear con fines cuantitativos y en estudios cinticos y termodinmicos. Algunos ncleos atmicos sometidos a un campo magntico externo absorben radiacin electromagntica en la regin de las frecuencias de radio o radiofrecuencias. Como la frecuencia exacta de esta absorcin depende del entorno de estos ncleos, se puede emplear para determinar la estructura de la molcula en donde se encuentran stos. Para que se pueda emplear la tcnica los ncleos deben tener un momento magntico distinto de cero. Esta condicin no la cumplen los ncleos con nmero msico y nmero atmico par (como el 12C, 16O, 32S). Los ncleos ms importantes en qumica orgnica son: 1H, 13C, 31P, 19F y 15N. Otros ncleos importantes: 7Li, 11B, 27Al, 29Si, 77Se, 117Sn, 195Pt, 199Hg, 203Tl, 205Tl, 207Pb

Informacin obtenida mediante RMN

La aplicacin fundamental de la espectroscopia de RMN es la determinacin estructural, ya sea de molculas orgnicas, rgano-metlicas o biolgicas. Para ello es necesario la realizacin de diferentes tipos de experimentos de los cuales se obtiene una determinada informacin.
Para la elucidacin estructural de molculas orgnicas y rgano-metlicas los experimentos ms utilizados son los siguientes:

Desplazamiento qumico

Indica el tipo de protn que provoca cada seal

Espectrmetro de masas
La espectrometra de masas es una tcnica de anlisis que permite la medicin de iones derivados de molculas. El espectrmetro de masas es un instrumento que permite analizar con gran precisin la composicin de diferentes elementos qumicos e istopos atmicos, separando los ncleos atmicos en funcin de su relacin carga-masa (z/m). Puede utilizarse para identificar los diferentes elementos qumicos que forman un compuesto, o para determinar el contenido isotpico de diferentes elementos en un mismo compuesto.

El espectrmetro de masas mide razones carga/masa de iones, calentando un haz de material del compuesto a analizar hasta vaporizarlo e ionizar los diferentes tomos, el haz de iones produce un patrn especfico en el detector, que permite analizar el compuesto. En la industria es altamente utilizada en el anlisis elemental de semiconductores, bio-sensores y cadenas polimricas complejas. Drogas, frmacos, productos de sntesis qumica, pesticidas, plaguicidas, anlisis forense, contaminacin medioambiental, perfumes y todo tipo de analitos que sean susceptibles de pasar a fase vapor e ionizarse sin descomponerse.

INVESTIGACIN PRECLNICA

Los experimentos que se realizan para el estudio de los efectos de los frmacos se llevan a cabo en animales de laboratorio, ya sea en el animal ntegro o en alguno de sus rganos, por lo que los experimentos pueden ser de tres tipos: a)= IN VIVO, cuando se utiliza al animal ntegro, mantenindolo vivo, para observar y registrar los efectos de los frmacos.

b)= IN SITU, cuando se utiliza alguno de los rganos o tejidos del animal, exponindolo por ciruga en el sitio anatmico correspondiente, para lo que se requiere que el animal est anestesiado, desmedulado y/o descerebrado.
c)= IN VITRO, cuando se efecta el experimento en una muestra de un rgano o tejido que fue extrado de un animal el cual fue previamente sacrificado, manteniendo dichos tejidos en condiciones de temperatura y nutricin similares a las fisiolgicas.

Bsicamente el avance de los conocimientos cientficos mdicofarmacolgicos dependen inicialmente de la experimentacin en animales de laboratorio, por lo que se hace necesario el empleo de una gran variedad de ellos, considerando que dentro de las normas legales de la investigacin farmacolgica preclnica se establece que los nuevos frmacos deben someterse a la experimentacin en por lo menos tres especies de animales, aunque el tipo de animal a utilizar depender de la rama o rea de experimentacin de que se trate. En los experimentos farmacolgicos dentro de un laboratorio, las especies animales ms utilizadas son los vertebrados que por su costo, su fcil y rpida reproduccin, as como su tamao proporcionan comodidad en su manejo; estas son: SAPO, RATON, RATA Y CONEJO.

VIA INTRAVENOSA: En el conejo se elige la vena marginal de la oreja, en donde se inserta la aguja con el bisel hacia arriba. En ratas y ratones se puede utilizar la vena marginal de la cola. VIA INTRAPERITONEAL: Tomando en cuenta la rpida absorcin por esta va y el fcil acceso a la misma, es una de las vas ms utilizadas en el laboratorio. En el caso del conejo, se toma por el dorso, se vuelve hacia arriba presentando la regin abdominal, se sujeta firmemente de las patas posteriores y se inyecta en la parte alta del cuadrante inferior izquierdo del rea abdominal, insertando la aguja con una inclinacin de 45 grados con respecto al plano corporal. En el ratn y la rata, se expone la regin abdominal y se inyecta en el cuadrante inferior izquierdo; la aguja, de 27 X 6 mm, debe formar un ngulo de 10 grados con el plano corporal. VIA INTRAMUSCULAR: En el caso de esta va, se presenta el dorso del animal y el frmaco se deposita con una aguja de 27 X 13 mm en la parte posterior de los cuartos traseros. VIA SUBCUTANEA: El frmaco es depositado por debajo de la piel del dorso con una aguja de 27 x 6 mm, levantando la piel con una mano e introduciendo la aguja con la otra.

RGANOS AISLADOS En la bsqueda de receptores especficos que median la accin observada tras la administracin de frmacos en el animal entero, as como su potencia relativa en comparacin con otros agonistas o antagonistas, el farmaclogo comenz a utilizar la tcnica del rgano aislado como pieza ineludible en el estudio del mecanismo de accin de los frmacos. La utilizacin del rgano aislado como reactivo farmacolgico aporta varias ventajas con respecto a los preparados in vivo: 1- Posibilita la cuantificacin precisa de la respuesta o efecto 2- Conocimiento exacto de la concentracin del frmaco sin interferencias de procesos de acceso y disposicin. 3- Eliminacin de respuestas de carcter reflejo (neuronal) o retroalimentacin humoral. Sin embargo tambin posee sus limitaciones ya que la tcnica exige una gran minuciosidad y un correcto montaje del preparado. A pesar de intentar mantener las condiciones fisiolgicas, debemos tener en cuenta que el rgano va a estar en un medio artificial.

Se deben tratar de respetar los siguientes parmetros: 1-Composicin electroltica 2-Osmolaridad 3-pH 4-Presin parcial de O2 y CO2 5-Fuente de energa (metablica) 6-Temperatura -Tensin mecnica Como fuente de energa se utiliza la dextrosa y ms raramente otros sustratos como sucrosa o aminocidos.

DISEO DE FORMAS FARMACUTICAS

Matricaria recutita Presenta una inflorescencia en forma de captulo paniculado. Las flores radiales son unos 20 mm, con la lgula blanca (ptalos), mientras que el receptculo central contiene flores amarillas, tubulares y numerosas, hermafroditas; el receptculo es hueco y carece de escamas, lo que permite distinguirla fcilmente

FORMULACIONES A PARTIR DE MANZANILLA (Matricaria)

NOMBRE CIENTFICO: Matricaria recutita L. = Matricaria chamomilla L. Asteraceae

NOMBRE COMN: Manzanilla

PARTE TIL: Las flores COMPOSICIN: Los captulos florales contienen aceite esencial (0.2-1.8%), constituido principalmente por: camazuleno, (-)- -bisabolol, ter cclico poli-eno, ino, 1.8-cineol y diversos hidrocarburos. La planta contiene flavonoides: apigenina luteolina y quercetina y derivados; cumarinas: dioxicumarina, herniarina y umbeliferona; carotenos; vitamina C; cido saliclico y esteroides derivados del estigmasterol, apina, jolina y fitosterina. Tambin contiene Lactonas sesquiterpnicas (matricida y matricarina) y polisacridos.

PARMETROS DE CALIDAD DE LA DROGA

PARMETRO

OMS,1991

NRSP 317

NORMA PROVISIONAL

HUMEDAD (%) CENIZAS TOTALES (%) CENIZAS INSOLUBLES EN HCl SUSTANCIAS EXTRAIBLES ETANOL 50% SUSTANCIAS EXTRAIBLES EN AGUA ACEITES ESENCIALES

No > 12 % No > 13 % No > 4 % No< 0,4 %

No > 13 % No > 12 % No < 30 % No< 0,4 %

11,26 1,645 31,68 8,203 -

Base universal (inerte) Manteca de cerdo o res..33 g Etanol 70 %........................................33 mL Cera de abejas o de caa.4 g Almidn de maz.10 g Agua aromtica0,4 mL Agua destilada.100 mL La manteca y la cera se disuelven en 80 mL de agua y 33 mL de etanol al 70 %. Se aade el almidn disuelto en los 20 mL de agua restantes poco a poco y con agitacin constante. La mezcla se deja enfriar y se aade el agua aromtica. Se mezcla y se guarda en refrigeracin.

Base para cremas.

a. b. c. d. e. f. g. h. Alcohol estearlico cido esterico.17 g Propilen glicol10 g Metil parabeno0,25 g Propil parabeno..0,03 g Petrolato slido, cera blanca vaselina slida20 g Petrolato lquido3 g Polisorbato 803 g Agua destilada42 mL

Paso 1. Se funden en bao de agua (a), (e) y (f) Paso 2. Se disuelve el resto de los ingredientes en el agua calentando. La disolucin del paso 1 se vierte sobre la del paso 2 poco a poco y agitando constantemente hasta solidificacin.

Extracto fluido. Se realiza por percolacin utilizando etanol al 50 %

PARMETROS EXTRACTO
PARMETROS ORGANOLPTICO

DE

CALIDAD

DEL

NRSP 324 Lquido pardo amarillento, transparente em capa fina, de olor caracterstico 5,0 6,0 0,970 1,030 1,365 1,385 No< 10 % No< 35 % No< 0,2 %

pH DENSIDAD NDICE DE REFRACCIN SLIDOS TOTALES CONTENIDO ALCOHLICO ACEITE ESENCIAL

Garanta: Tres aos

Crema al 10 % Base universal inerte.100 g

Uso: Cosmtico para proteger y suavizar la piel

Dosificacin: Aplicar dos veces al da Garanta: Un mes

PARMETROS DE CALIDAD DE LA CREMA PARMETROS ORGANOLPTICO NORMA PROVISIONAL Crema homognea, sin arenosidad, color verdoso y olor caracterstico 5.67 0,26 1.36344051 0.00226925

pH INDICE DE REFRACCIN

Jarabe al 10 % Extracto Fluido de Manzanilla. 10 mL Metil parabeno. 0,18 g Propil parabeno 0,02 g Alcohol etlico 1 mL Jarabe simple c.s.p...........................................................100 mL Nota: Puede sustituirse el metil y propil parabenos por benzoato de sodio 0,5 % pH 5,0 - 6.0 Disolver el metil y propil parabenos en alcohol etlico e incorporarlo al extracto fluido. Adase lentamente el Jarabe Simple hasta completar volumen. Uso: Se utiliza en el tratamiento de los trastornos digestivos y trastornos nerviosos como depresin, ansiedad e insomnio. Dosificacin: 1 cucharada 3 veces al da. (1 cucharada equivale a 8mL/ administracin) Garanta: 6 meses

Champ

Extracto fluido.10 mL Cloruro de amonio3 g Lauril sulfato de sodio3 g Miel de abejas, glicerina o propilen glicol3 mL Agua destilada csp..100 mL Se mezclan todos los ingredientes y al final el espesante lauril sulfato de sodio Uso: Anti seborreico y antisptico del cuero cabelludo Dosis: Usar una vez al da Garanta: 3 meses

PARMETROS DE CALIDAD DEL CHAMP

PARMETROS pH

NORMA PROVISIONAL 5,69 0,455

DENSIDAD
NDICE DE REFRACCIN

1,01290 0,104
1,3510 0,005

PROPUESTA METODOLGICA PARA LA INVESTIGACIN INTEGRAL DE RECURSOS NATURALES TERAPETICOS

Teniendo en consideracin los mltiples factores que inciden en la problemtica de salud y considerando que las plantas medicinales representan un valioso recurso, que explotado racionalmente y previo un estudio integral constituiran una alternativa de solucin a los problemas de salud, se plantea una propuesta metodolgica para la Investigacin Integral de Recursos Teraputicos, la misma que por la complejidad del problema que pretende enfrentar puede ser desarrollada e implementada por mdulos, con las consiguientes; ventajas:

- Sistematiza las experiencias populares del uso de plantas medicinales desarrollando el mtodo cientfico. - Identifica reas crticas de investigacin aplicada, ligndolas a un enfoque participativo y estratgico. - Simplifica las etapas del mtodo cientfico, dndole un carcter dinmico y acadmico. - Permite obtener resultados que puedan servir para implementar polticas de salud. - Incorpora mtodos de salud pblica en investigacin.

MODULO 1: FORMACIN DEL EQUIPO INTERDISCIPLINARIO DE INVESTIGACIN. Hoy en da las investigaciones cientficas necesitan de la integracin de diversos grupos de trabajo que tributen a un mejor desarrollo y conclusiones del proyecto propuesto para lo que se desea investigar y poder lograr la solucin del problema planteado. MODULO 2: DIAGNSTICO, PRIORIZACIN DE PROBLEMAS Y PLANTEAMIENTO DE ALTERNATIVAS EN SALUD. Constituido el grupo interdisciplinario se organiza la propuesta teniendo en cuenta: - Diagnstico situacional de salud de la regin. - Problemtica del medicamento y porque es necesaria la investigacin como solucin alternativa. - Ejercicio de la Etnomedicina y como la informacin sobre su uso ayuda al desarrollo de la investigacin planteada. - Metodologa de Investigacin en Plantas Medicinales. Concepcin del proyecto

Realizado el Diagnstico situacional se procede a priorizar los problemas de salud y al planteamiento de alternativas dentro del marco de las siguientes interrogantes: En qu medida las investigaciones actualmente desarrolladas responden a las necesidades prioritarias de salud de la poblacin y cules son los factores que han intervenido positiva o negativamente? Cules son las problemticas que han motivado la investigacin actual? Con esta investigacin se darn propuestas para la posible produccin y/o aplicacin de las alternativas para la solucin alternativa de los problemas de salud planteadas. Las conclusiones y recomendaciones generales deben incluir dos aspectos fundamentales: Encuesta sobre el uso de plantas medicinales en la regin y como ayuda al sistema de aplicacin de los medicamentos de ms frecuente uso en la terapia habitual.

MODULO 3: DESARROLLO INTEGRAL DE LA ETNOMEDICINA Identificados los problemas ms prevalentes de salud y como alternativa viable para la Regin, la Etnomedicina, se procede a la seleccin de plantas medicinales a investigar tomando como criterios: 1. Uso en problemas ms prevalentes de salud. 2. Frecuencia de uso popular 3. Accesibilidad de cultivo 4. Compilar los resultados tcnicos y cientficos reportados 5. Realizar un listado de plantas medicinales en orden prioritario de utilizacin. 6. Compilar la informacin especfica sobre las plantas medicinales priorizadas, a dos niveles: - Bibliogrfico: En el mbito local, nacional e internacional, va base de datos. - Sistematizando la experiencia popular: en base al uso en enfermedades ms prevalentes y la prctica popular de terapia con plantas medicinales, sistematizacin que concluye con la identificacin y clasificacin taxonmica de las Plantas Medicinales.

MODULO 4: INVESTIGACIN CIENTFICA Y EPIDEMIOLGICA. Disponiendo de la informacin pertinente se procede a la investigacin cientfica va el anlisis fitoqumico en un primer nivel por grupo qumico y por especie vegetal. Luego se procede a la evaluacin farmacolgica de los grupos qumicos, identificndose aquellos que muestren actividad farmacolgica "in vitro" o "in vivo" (animales de experimentacin). Realizar el estudio fitoqumico hasta la identificacin de la molcula responsable de la actividad teraputica, la misma que ser sometida nuevamente a evaluacin farmacolgica para confirmar su actividad. Paralelamente a este estudio se realiza la investigacin epidemiolgica teniendo en cuenta dos aplicaciones metodolgicas que son: 1. La Vigilancia Epidemiolgica de los casos que sean manejados con plantas medicinales, mediante monitoreo clnico, y 2. El enfoque de riesgo de los grupos poblacionales ms susceptibles priorizados para el estudio, segn la patologa de la regin.

MODULO 5: ELABORACIN DE PROTOCOLOS Y EJECUCIN. Si los resultados de la investigacin sugieren la aplicacin de los resultados, entonces se formulan los Protocolos para las siguientes reas de investigacin: 1. Propagacin de cultivos 2. Preparaciones galnicas 3. Distribucin y comercializacin 4. Seguimiento sistematizado Se procurar mantener la misma metodologa asumida en los mdulos anteriores. La ejecucin de los mismos se desarrolla en forma coordinada y paralela entre los grupos de investigacin especializada.

MODULO 6: APLICACIN DE RESULTADOS Y EVALUACIN DE LA METODOLOGA. Se realizar la bsqueda de acceso a los sistemas formales de salud con aquellos resultados de la investigacin que sean vlidos, adems se difundir los datos a los grupos interesados del nivel popular, tcnico y cientfico, con la finalidad de recoger sus apreciaciones y sugerencias. Finalmente se evaluar la metodologa propuesta con la finalidad de reforzarla y hacer el uso apropiado de la misma. Se medir el IMPACTO en la solucin de los problemas de salud para los cuales se hizo la investigacin en la conservacin ecolgica para evitar la depredacin de plantas medicinales y su influencia en la economa de los grupos beneficiarios de las investigaciones.

 Ttulo Autor. Centro de trabajo. Tutor(es) y/o investigador principal responsable del proyecto, grado cientfico, centro de trabajo Conocimiento previo. Breve fundamentacin bibliogrfica con sus citas correctamente acotadas.Importancia de los resultados a obtener

GUA PARA LA CONFECCIN DE UN PROYECTO.

 Problema Hiptesis Objetivo general

Objetivos especficos
Cronograma de tareas, con fechas de cumplimiento Descripcin de mtodos por tareas, incluyendo el anlisis estadstico en los casos que proceda.

GUA

 Recursos disponibles para el desarrollo de la investigacin. Recursos necesarios para completar las necesidades materiales. Necesidades financieras y su utilizacin. Balace econmico total de la propuesta Citas bibliogrficas referidas.

GUA