Dec 9, 2010

“LASHUNA SHODITHA PARADA--- A PROCESS STANDARDISED LEAD MOLECULE FOR A NON-TOXIC MERCURIAL THERAPEUTIC DRUG”

 Abstract:-
Key words: Alliin alliicin, allinase enzyme, methyl cysteine, ajoene, antidote,
                    theosulphinate,lachrymatory-synthase,sulphoxide(methiine),
                   L-canavanine,apoptosis,cytochrome p-450,glutathione.


A human act or activity for adjustment,readjustment ,tuning ,normalization,stylization,social control,stabilization is what is meant as standardization.

Acharya charaka mentions 64 minerals which included metals such as Cu,Ag,Au,Pb,Zn,Sb,Fe,Ar etc..the tradition of processing metals and metallic products for medicinal use was well known to our acharyas who were aware of its detrimental effects.unlike western medicine Ayurvedic medicines are based on years of experience ,observations,empiricism and intution and has been handed down to generations both through words and treatises.
Safety,efficacy,standardization,quality control and regulations of the raw materials, manufacturing process and the ultimate goal is the finished medicinal product.though standardization of minerallo-metallic,herbo-mineral drugs is much discussed but is little understood an expression of standardization of each and every process is needed and is one of essential part of non-toxic standardization of herbo-mineralo-metallic drugs.
The shodhana process of minerals and metals is one such process which is widely descripted in all classical texts that can be interpreted in the light of modern biochemical,organic,inorganic,systemsbiology,pharmaceuticalscience,stoicheometry,medicinal chemistry etc..
Here is one such rationalized scientific approach in standardizing the purification procedures of mercury that would result in action on targeted cells.

Garlic (alliicin-organosulphur) and Hg reaction is a redox process where there is a reactant undergoing oxidation and one undergoing reduction. Since the two reactions are complementary to one another and occur simultaneously. The electron changes in oxidant and reductant forms the basis of ion-electron method for balancing ionic equations. There may be molecules of H+ and OH­- ; all may be used in balancing the partial equations.

The sulphur and mercury forming a best co-valent bond within garlic and mercury and as a result the triturated product turns black in color though may of mercury salts are turned to white powders or crystals. It is a miniature concept of Kajjali (mercuric sulphide) itself.

So the drug designers of ancient time have proposed parada samanya Shodhana using garlic. Even though we are unable to trace the reason for selecting garlic in samanya shodhana of parada in original text with the advanced photochemical research and other allied researches will encourage the scientific thinking of parade shodana. Ofcourse prospective instrumental research based on this critical literary research will only be confirmative objectively.
Amongst various parada shodhana methods garlic trituration method is also ideal and applicable as garlic is antidote for heavy metal poisoning, contains sulphur and hence an innate codrug for parada bandha, chemically sulphoxide, thiosulphinates, methyl cystiene, L-canavanine, ajoene etc are significant phyto constituents of organic nature helping inorganic mercury to end in safe absorbable pharmacotherapeautically effective mercury molecule.

Article:-

Objective:
To find the scientific explanation for samanya parada shodhana (the lead molecule) by lashuna trituration method to get the standardized medicinal product.(Rasatarangini, Ayurveda Prakasha).et al

Materials and Method:

Material:
Mercury, garlic and the literature regarding the same.
Method:
Retrospective and prospective logical scientific literary research.

                                            DETAILED DESCRIPTION

CONCEPT OF SHODHANA

CORROSION: corrosion of metals is their destruction under the action of surrounding media such as water, steam & furnace, gas etc as a result chemical and electrical process that takes of place on the surface of metals which may be in different forms uniform/point /honey/comb./intercrystalline.
A passive oxide film formation is the process in which metal /alloy exhibits a much higher corrosion resistance than expectance from its position. this protective layer is the one which makes the metal noble.(ion selective electrodes are an ideal measurement tools to know the ion activities which are important parameters to predict the corrosion rates ,solution conductivities ,effectiveness of metal parking baths and physiological effects of ion in biological fluid .

HEAT TREATMENT: fire was a mystical force that transformed one substance to another and was thus an object of wonder and superstition .it was fire that led to the discovery of purification of metals which gave rise to metallurgy.

WHY HEAT TREAT METALS?
Ø    To optimize the mechanical and physical properties for a given application and make way for next chemical reaction /electro chemical reaction.
Ø    It is not a process of hardening metal but to give the increased strength and wear resistance.
Ø    Controlled heating, cooling of metals alter the properties, increases strength and as a result increasing the product performance.

SURFACE TENSION: there is an interfacial tension which is biochemical very important especially very important in the process of adsorption height and density of a substance is also very important ,as the molecules of the surface are not so free to move .they are held together and for a membrane over the surface of a liquid .this force with which the surface molecules are held together is called surface tension .the surface tension decreases and increases with temperature .organic substances usually have low surface tension in solid to semi-solid to liquid systems the immersed substances lower the interfacial tension.
MASS AND ENERGY:
In all reactions atoms are molecules (chemical reactions) or between nuclei (nuclear reactions) there is a conversion of mass into energy or the converse according to whether energy is given out or absorbed during the reaction,the mass changes occurring during chemical reactions are so small as to be immeasurable but in nuclear reactions they are large enough to be detected .the major pathway of oxygen use in aerobic organism is 4 -electron reduction to give two molecules .
O2+4H-+4e -   ------2 H2O Eo =+0.815V, a major source of energy.
  

Aerobic purification:
Oxygen carries a lot of magnetic charges that affect largely the miracles of any process going around. The open trituration method in a khalva would allow this atmospheric oxygen to react with in a self created magnetic field. Mercury interferes with the capacity to quench highly reactive oxygen species to form inorganic mercury compounds.

Purification due to trituration:
Mardanam gunavardhanam. Let us analyze what are the qualities procured during trituration. The first entity would be rising temperature.

Rising temperature:
The heat produced due to friction of the pestle and mortar along with uniform rising temperature (if tapta khalva) would seize the heat irresistant impurities. Also, the rise in temperature accelerates an enzyme reaction but at the same time causes inactivation of the enzymes due to denaturation of the proteins. At a certain temperature known as optimum temperature the activity is maximum. However the optimum temperature of plant enzyme is 600 C. If the temperature is lowered, the rate of enzyme reaction is diminished. At temperature 00 C most enzymes are practically inactive and are destroyed by moist heat at 1000 C.

GARLIC (AJO-ALLIUM SATIVUM-liliaceae)
Garlic is one of the earliest known medicinal plants .its cloves had been used as a cure all in ancient Egypt .in Ayurveda garlic is recommended against leprosy.
active principles-sulfur containing compounds Alliin is converted to the antimicrobially active allicin,when the bulb is cut or bruised.Ajoene which is a secondary  degradation product of Alliin ,is presumably the most active compound

MERCURY:
 Mercury as a native metal its boiling point is around 357 0 C .
Specific gravity -13.59.
Hg combines with most of the metals to form alloys called amalgams. Its atomic number being 80 and atomic weight is 200.59. To get more yield of mercury today there are many sophisticated methods technically to produce pure mercury. Chamois leather used for physical insoluble impurities and HNO3 for chemical treatment. Hg thus obtained is subjected to vacuum distillation so as to get 100% pure form.

Even though 100% pure Hg is obtained by this or similar such methods, such Hg cannot be and should not be used to prepare medicines but should and must undergo shodhana classically.


Action of crushed garlic:
Garlic (Allium sativum) belonging to liliaceae is probably the earliest known medicinal plants.the fresh bulb contains alliin,allicin and volatile oils.when the garlic is crushed, alliin the first organosulphur compound found in nature to display optical isomerism is exposed to enzyme alliinase and converted to alliicin within 10 seconds .Alliicin is unstable and converts readily into mono, di, tri, and poly sulphides, sulphur oxide and other compounds such as ajoene, which is a secondary degradation product of alliin is presumably the most active compound responsible for any multiple bonding along with mercury.

Hg+Ajoene(Sulphur oxide)   leads to     Mercuric sulphuroxide

It is the enzymes which are free to react with the active sites to induce a configurational change (induced fit mechanisms). The presence of metalloenzymes in a definite quantity for the function of metal ions is retained throughout the purification process. These metal activated enzymes bind metal less tightly. But here in this process metal ions act as catalyzers for co-valent bonding.
     Due to enzymatic degradation of sulphur containing amino acids, as the process goes on it forms cysteine sulphoxide lyase which is known to represent important fraction of total protein content in allium bulbs.
The bonding structure found in Alliin, the major sulphur compound in whole, unbruised garlic.

                                                O

H2C = CH- CH2                                               C - OH
                               S  -CH2 -C
                                                    NH3
                    O


In allicin the next organosulphur compound produced after crushing due to enzyme action represents a different pattern of bonding.

H2C = CH - CH2

                                  S - S
                     O
CH2 - CH=CH2                       




Once formed alliicin is unstable, pure alliicin at room temperature has a half life of 2-16 hours, if in garlic juice or in crushed garlic 2-4days. As the enzyme alliinase and lachrymatory- factor synthase substance are present in separate compartment apart from amino acid substance in vivo. Upon rupture or wounding of the cells the enzyme located in the vacoule and the alkyl (methyle) cysteine sulphoxides located in the cytoplasm of allium can react and produce the volatile and odorous compound. Therefore alliicin which is derived from alliin is further classified as theo sulphinate (2S atoms bonded together one of which is bonded to O2 atom. This alliicin accounts for about 70-80% of total thiosulphinates in garlic. To get the maximum amount of alliicin from allin the bruised garlic should be used as soon as possible.


BIOLOGICAL PATHWAY

The use of oxygen in aerobic organisms is to function as a source of oxygen atoms in the biosynthesis of various molecules and conversion of lipid soluble to water soluble forms. The genetic configuration of garlic and its reaction bonding with Hg makes it more potential to be a purified mercury.

Alliicin is antimicrobially, antifungally, antibacterially active due to the concentration of sulphur compounds. Apparently allinases are translated from mRNA species of approximately 2200 nucleotides. The primary translation products are preproteins which are converted into the mature alliinases following post translational modifications. In garlic alliinase the mRNA encodes a 486 aminoacids polypeptide with a molecular mass of 55623 Daltons. The protein sequencing which is conducted on bio system model 477A protein sequence shows young shoots, old cloves, young shoots with shallots, old bulbs of garlic with different c DNA sequence. It is shown that garlic allinase expressed in all levels of old and young bulbs of same and alliinase is present through the development of plant. Therefore selection of garlic cloves for trituration does not make large difference.
Fresh garlic juice exhibit high broad spectrum antimicrobial activities .It alliin and alliicin also involved in the proven antioxidant effect of garlic. But this effect is dose  dependent and excessive concentration of alliicin act as pro oxidant.

Hence forth in our classicals the quantity of Bhavana/mardana dravyas is said as

            " Dravena Yavatha dravyam choornitham thu ardrathaam vrajeth 
             Taavaneva dravo dravye deyo bhishghbhirbhavanaavidhou "


The liquid to the powder in optimum proportion, that it gets sufficient moisture and forms a pulp. Alliicin has a low solubility in water 2% where it degrades to sulphides including di,tri,methl trisulphide which are also physiologically active as well.

Garlic extracts, through their antioxidant activities have been reported to provide protection against free radicle damage in the body. Anti oxidant properties of allicine, allylcysteine & allyl disulphide prepared by chemical synthesis or purification. Allicine scavenged superoxide , while allyl cysteine4 and allyl disulphide will not react with superoxide. Allicine and allyl cystein are hydroxyl scavengers. Allyl disulphide is a lipid peroxidation terminator. In summary all these exhibit different patterns of antioxidant activities as protective compounds against free radical damage. The large amounts of chlorophyll,
Beta carotene, vitamin-E, amino acids, L-canavanine which has anti bacterial, antiviral effect along with stimulation of interferon’s which enhances the natural killer cells.
As garlic extracts or antioxidants they provide protection against free radical damage. All patterns of alliicin and cysteine are protective compounds against free radical damage. The anti biotic, antibacterial and anti viral properties of garlic extract is due to L-canavanine which stimulates the interferon production, enhancing the natural killer cells.


ELECTROMAGNETIC PURIFICATION:


REDOX REACTION:

Attractive forces necessary to produce coherence between molecules that is responsible for the liquid and solid states. if repulsive forces did not operate there would be nothing to prevent the attractive forces from causing interpretation and mutual destruction of molecules.

The Hg polarography studies show-The magnetic circuit provided in the electrode and anode reactions when mercury is immersed in solution of elecrolytically active species, due to its relatively large surface area, has a low current density at its surface and is non polarized thuds remaining at constant potential. The pH of mercury been 6 is nearer to acidic nature ad the catalyses of the garlic base would make a strong co-valent bond of Hg++ with cations of sulphur compounds. Hg++ acts as a ‘super acid’ to form pi bonds . Unlike protons metal mercury can serve as a three dimensional template for orientation of basic groups on the enzymes or substrates.
The co-ordination sphere of mercury may bring together enzyme and substrate are form chelate producing distortion in   either  the enzyme or substrate. Mercury may also mask a nucleophile and thus prevent side reactions.

RADIOACTIVITY:
.A study says the effect of an acute dose of n - rays ( 10 gy) to post dormant garlic cloves on inner sprout growth and changes in peroxidases and soluble proteins were evaluated up to 100 days of storage in darkness at 19-10o C and 42-2% relative humidity. Radiation induced inhibition of sprout growth became evident after 25 days of treatment and was synchronous with marked increase in the number of anodic peroxidase, iso enzymes at 100 days. Neither the soluble protein nor the protein pattern was affected by irradiation. These are in terms of possible mediating effect of peroxides of radiation induced sprout inhibition in garlic. This shows that even the irradiation effect of mercury while trituration is suppressed and neutralized by garlic.


ELEMENTOLOGY IN PURIFICATION:

A give and take principle is followed between mercury and garlic when all the other factors are intact.
The selenium and Germanium trace elements found in the garlic extract is another area of explanation showing some unknowing effect on Hg. Because Hg forms a tight bond with selenium there by displacing selenium from its critical role as an obligatory constituent of  glutathione  peroxidase.

PARINAMA-On transformation matter is constantly undergoing change of state. The effect is only the cause in the new collocation…

TOXICOLOGY:

C-hg bond of some organic hg is cleaved after absorption, with methyl mercury the cleavage is quite slow and the inorganic mercury formed is not thought to play a major role in methyl mercury toxicity.

Garlic has been proved as a best antidote for heavy metal poisoning. Hence processed garlic is augmented with antidote itself. Hence one step ahead in safety to select it as a best drug for shodhana of Parada and logically used.

CHEMISTRY:

The thermodynamics, kinetics and free radicals auto oxidation
 Process is the basis of the chemistry of metals. A molecular species in the reaction medium may combine with one in the reaction mixture modifying the kinetics of the reaction.
Hg readily forms co-valent bonds with sulphur and it is this property that accounts for most of the biological properties. The affinity of mercury for ‘thiols’   provides the basis for treatment of mercury. The methyl cysteine thiosulphate combines with mercury to form methyl mercury which can be safe and readily absorbed by the human GI tract in inorganic form…

CLINICAL PHARMACOLOGY:

Khalvi rasayanas are the basic products for any formulation of the drugs further. Though parada Samanya shodhana is said to be done with many drugs such as Haridra, kumari, Chitraka etc.Pharmacological path is activated or formulated keeping in mind the targeted tissues and so parad shodhana with lashuna is simulation to give the result of such an end product which can eradicate the root of the disease.
As a merit lashuna containing sulphur as parada always as affinity towards sulphur groups disintegrated, conjugated and form as one unique compound hg-s which presents as a Lead molecule.

Ajone inhibits thromboxane synthesis through the inhibition of cyclo oxygenase and lipo oxygenase enzymes. For the treatment of pulmonary syndrome garlic is proved a s a good drug.

The processed triturated mercury with garlic in purification can be used in formulating such drugs which can be indicated essentially in CADs, CVDs, hyperlipedimiea, tumours, hormonal disorderts, atherosclerosis, obesity. Hence the purification methods of hg with garlic would be unique with such pathologies. As garlic possess quality of killing disease, it would be more effective. Eg:- classically formulated drug such as Chandrakala rasa18 which is indicated in hypertension where in parada is one of  main ingredients can be purified by using garlic for both the cause of detoxification and enhancing its qualities. Bhruhat garbha chintamani rasa that is used as rasayana along with prevention of giddiness and other risks in pregnancy also contains parada, which can be best purified by lashuna. Gulma kalanala rasa containing parada is another example, which can be best indicated in oncology. Arogyavardhini vati a common OTC drug with its broad spectrum of indications is known to be sroto vishodaka, kleda and Medho hara. Parada in this act as lead molecule that can be processed with lashuna for its better efficacy.
Because all in all main objective of performing proper procedures and formulating the drug is nothing but for the ultimate aim of treating the disease.

DISCUSSION
Garlic (alliicin-organosulphur) and Hg reaction is a redox process where there is a reactant undergoing oxidation and one undergoing reduction. Since the two reactions are complementary to one another and occur simultaneously. The electron changes in oxidant and reductant forms the basis of ion-electron method for balancing ionic equations. There may be molecules of H+ and OH­- ; all may be used in balancing the partial equations.

The sulphur and mercury forming a best co-valent bond within garlic and mercury and as a result the triturated product turns black in color though may   of mercury salts are turned to white powders or crystals. It is a miniature concept of Kajjali (mercuric sulphide) itself.
So the drug designers of ancient time have proposed parada samanya Shodhana using garlic. Even though we are unable to trace the reason for selecting garlic in samanya shodhana of parada in original text with the advanced phytochemical research and other allied researches will encourage the scientific thinking of parada shodhana. Of course prospective instrumental research based on this critical literary research will only be confirmative objectively.

CONCLUSION:-
Amongst various parada shodhana methods garlic trituration method is an ideal and applicable as garlic is antidote for heavy metal poisoning, contains sulphur and hence an innate codrug for parada bandha, chemically sulphoxide, thiosulphinates , methyl cystine, L-canavanine, ajoene etc are significant phyto constituents of organic nature helping inorganic mercury to end in safe absorbable pharmacotherapeautically effective mercury molecule.

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