Describes the research work done by Dr. D. C. Sharma, Prof. of Biochemistry on trace elements - silver, gold, mercury, iron, strontium, copper, Calcium, Lead, Selenium, etc.

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EXPLORING INORGANIC BIOCHEMISTRY
( Biochemistry of Some Inorganic Elements)
By Dr. D.C. Sharma
KEY WORDS : Inorganic Biochemistry, Heavy elements, Trace metals,
Iron, Noblemetals, Silver, Gold, Mercury, Copper,
Strontium, Calcium, Lead, Selenium
Inorganic Biochemistry is a broad term which encompasses the study of
occurrence, functions and effects of inorganic elements or their compounds in
biological systems . This is traditionally and more commonly known as the study
of ‘trace metals’ or ‘trace elements’. Both of these terms are quite vague and
restrictive.
I started research in 1966 . Iron deficiency anemia (IDA) was a serious
public health problemat that time which continues to be so even today.
Gradually other elements werestudied Thus iron, mercury, silver, gold, copper,
and strontiumwere investigated .
Any discussion of inorganic elements would be incomplete without
reference to the periodic table of elements (Fig: 1) . We find that out of more
than 90 elements known to exist in naturethe living cell contains less than one-
third of them, i.e., around 30 and none of the element beyond iodine (at.no. 53)
has so far been found to have essential function. Future research workers should
concentrate on elements beyond iodine. They should specifically look for trace
element function of following elements.

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Fig. – 1 The Periodic Table of Elements
(a) Boron ( at.no. 5) as it is followed by a series of four essential
elements – carbon (C6)
, nitrogen ( N7
), oxygen (O8
) and fluorine(F9
) in the
second period . Itmay be mentioned that boron is considered essential for
plant growth.
(b ) Bromine (Br35
) as it is preceded by two essential elements –
fluorine (F9
) and chlorine (Cl17
) , and followed by another essential
element, iodine (I53
) in group 7a.
© Strontium (Sr38
) , whosetwo predecessors in group 2a ----
magnesium(Mg12
) and calcium (Ca20
) , are well known to have
essential function in the body.
(d) Aluminum (Al13
) is the only element yet to be discovered to have
any essential function in the third period, as all other elements
before it ---sodium( Na11
) and magnesium (Mg12
) and after it –

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silicon (Si14
), phosphorus (P15
), sulfur (S16
) , and chlorine (Cl17
) are all
essential elements.
India is a treasure-troveof material for research on inorganic biochemistry
as its ancient medical systems---- Ayurved (1,2) and Siddha (3) extensively use
metals and their bhasms (ash) for treatment of a variety of diseases and disorders
so these open a new and vastvista of research opportunities . Homeopathy (4)
also uses several inorganic compounds, but their concentration is too small to be
measured.
DIFFICULTIES IN TRACEELEMENTS RESEARCH
The research on inorganic elements or trace elements is fraughtwith
difficulties. Two types of problems are encountred. One is analytical, the other is
experimental. The routine biochemical analytical techniques, like, colorimetry,
spectrophotometry and fluorometry, arenot useful in inorganic trace element
research. For this purpose, one needs activation analysis, ion- selective electrode
or atomic absorption spectrophotometer (5) , or more recently developed
inductively coupled plasma (ICP) and mass spectrometry.
For analysis of inorganic elements, atomic absorption spectrophotometry
is much more common and universally applicable. Brief review of this technique is
published by me (5) and an excellent detailed review have also appeared (6).
The analysis of inorganic elements in biological samples by any technique,
except activation analysis and mass spectrometry , requires prior destruction of
organic matter and preparation of clear sampleof the analyte. This is usually
done by dry ashing or wet digestion. Both of these techniques have their own
advantages and disadvantages ; the mostimportant being contamination---
positive or negative (5). The problem of contamination has been discussed in
excellent review by Thiers (7) . The destruction of organic matter is discussed in
detail in a Monograph (8).
To investigate whether an element is essential, its deficiency has to be
firstproduced in an animal . Itis not an easy task as severalelements are required
in extremely small amount than usually present in the normal diet. For this

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purpose, Schwarz etal. (9) fabricated trace element – controlled isolators in their
laboratory which have proved very usefulin discovering newer essential trace
elements.
The animals are reared in these isolators and maintained on a special
purified diet, adequate in all known nutritional factors, including trace elements
so far identified as nutritional essential. The animals are closely monitored for
growth and any sign or symptomof deficiency , including changes in its
appearanceand behavior . Later on supplementation of the element under
investigation is done to the animal and its possibleeffect on reversing deficiency
changes are studied. If deficiency symptoms or changes are corrected it clearly
demonstrates essentiality of this element for that animal.
Using trace element controlled isolators , Schwarz and other investigators
indicated the essentiality of several elements. In somecases deficient diet had to
be given to next 2-3 generations of animals (usually rats) before getting proof of
essentiality.
The greatest contribution to our knowledgeof trace elements (essentialas
well as non – essential ) was made by Henry A. Schroeder (10) who relentlessly
reported his findings about one element after the other. To mention a few of
them : copper, zinc, magnesium, manganese, cobalt. nickel, chromium, lead,
strontium, bariumand many more. The prospectiveresearcher in inorganic
biochemistry is advised to go through his papers and two excellent treatise by
Underwood (11) and Mertz (12). For doing research in Inorganic Biochemistry,
the researcher should also have a good knowledgeof chemistry of elements. An
outstanding book has been recently published which gives a visualexploration of
all the hitherto known 118 elements (13).
Now, the summary of my research on different inorganic elements is
presented.
IRON
My research on iron was directed towards enzymic and metabolic
changes occurring in severe iron deficiency produced in rats by feeding a

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synthetic diet (14) and interaction of intraperitoneally injected colloidal
preparations of three metals – silver, gold and mercury with body iron (15). It
may be mentioned that it was for the firsttime that these metals in colloidal
formwere used in any biochemical experiment . This was the precursor of nano
particles of today.
The high incidence of iron deficiency anemia (IDA) in general population of
India consuming largely vegetarian diet, rich in iron, is due to a number of
substances presentin vegetarian diet which render iron unabsorbable(16). We
call it anemia of vegetarianism. In order to increase iron absorption fromthe very
same diet, a group of volunteers were given vitamin C tablet (500 mg) twice daily
after major meals. This was found to significantly increase hemoglobin, iron,
transferrin saturation and ferritin without any side effect, usually associated with
supplemental iron (17).
In order to tackle the problem of anemia and iron deficiency, another
study was designed to evaluate efficacy of six most commonly used Ayurvedic
iron – containing preparations (Navayasa Curna), PunarnavadiMandura, Dhatri
Lauha, Pradarantak Lauha, Sarv- Juara- Hara Lauha and Vrihat YakrdariLauha)
were given for thirty days to twenty IDA patients in each drug group and
compared with another group of 20 patients given best of Allopathic medicine
containing ferrous fumarate, vitamin C, folic acid and vitamin B12. The results of
hematologic and iron status parameters was better in mostof Ayurvedic
preparations and there was no side effect as observed with iron salts. The Hb
regeneration rate was 0.10 g/dl/day for Allopathic preparation while it was above
this value for all Ayurvedic preparations , except Pradarantak Lauha . Sarv-Juar-
Har -Lauha was the drug of choice as Hb regeneration rate with it was highestat
0.16g/dl/day (18). If itis so, it offers a great opportunity of eradicating iron
deficiency anemia from India using Ayurvedic systemof medicine. This will be
only possiblewhen physicians--- Allopathic as well as Ayurvedic do scientific
evaluation of Ayurvedic Medicines in their patients.
THE NOBLE METALS

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Certain metals are called “The Noble Metals”, the mostnotable of them
are gold, silver and mercury. Our work on the effect of colloidal solutions of these
three metals on iron metabolism has already been mentioned (15).
MERCURY : A lot of work was donein our laboratory on the toxicity and
metabolism of mercury which formed the basis of another Ph.D. thesis (19). Itis
not possibleto review its all findings here. Only two importantfindings are
mentioned.
Based on this work, a hypothesis was proposed and published about the
biochemical basis of toxicity of mercury (20). Theevidence was presented to
supportthe hypothesis that the toxicity of mercury is at least partly due to its
combination with coenzymeA and resultant interference in CoASH functions
(20,21).
Another important finding was the chance discovery of changein the
electrophoretic mobility of hemoglobin and albumin following mercury
administration in rats (22). Similar finding was also observed by us in humans
receiving gold therapy (23). It is believed that the altered electrophoretic mobility
of these blood proteins can be used for diagnosis of mercury or other heavy
metal poisoning or detection of their presencein blood.
SILVER: Silver is ingested by Indians in the formof varak (thin leaf),
Ayurvedic bhasma and water contained in silver utensils, but nobody knows its
effect, except as an anti- bacterial or water purifier. So we had conducted a study
by administering its varak , bhasma and colloidal solution (silver sol) to chicks to
see the effect on some essential elements (24). The results werenotable for
significant riseof silver in all tissues examined and significantfall in copper in
blood. This antagonistic relationship between copper and silver was expected
because they are in the same group (1b) of Periodic Table (Fig. 1) .Earlier
Hirasawa et al . (25) also reported significant decreasein copper in serumafter
injecting silver nitrate solution intraperitoneally in rats (25). Itis noteworthy that
the greatestrise of silver was observed in blood of chicks receiving silver sol,
which actually had the lowest amountof silver.

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In another similar study (26) these three silver preparations werefed to
another group of young male chicks for ten days. A significant fall in all the lipid
fractions of plasma __ total lipids, phospholipids, triglycerides and total
cholesterol, was observed. Therewas a marked rise in silver content of plasma,
and whole blood ranging from 4 to 13 times suggesting that the observed
hypolipidemic action was due to silver. The administration of these three silver
preparations did not causeretardation in growth, toxic manifestation, any side
effect or untoward reaction.
The hypolipidemic effect of silver was a very important finding. So it was
decided to confirmit in humans. For this purpose, silver leaf or varak was used
which is widely consumed in sweets and betels in India and in cakes and
confectionary in west for long time without knowing if it has some medicinal
value. In this study, about 50 mg of varak ( metallic silver) was given daily by
mouth to 30 healthy volunteers for 20 days. A statistically significant
hypophospholipidemic, hypotriglyceridemic, hypocholesterolemic and
hypoglycemic effect was observed (27). Thesafety of ingested silver was indicated
by absence of any abnormality in urine and unaltered levels of protein and
albumin in the plasma. Our observations suggestthatsilver could be beneficial in
conditions, like, diabetes mellitus, obesity and atherosclerosis. Therefore, my
advice to the to the prospectiveresearchers is to take up this projectfor further
study using different doses and duration of treatment in humans in these
conditions. Itmay provevery rewarding to them.
GOLD : Gold is the noblest of noble metals. Itmay be mentioned that gold
is highly valued in Ayurvedic systemof medicine in the formof varak (leaf) ,
bhasma (ash) and many other preparations of obscurechemical composition (1).
Apart fromthese, a wide variety of gold preparations areavailable commercially
in the market, even without prescription (28). Almostall of them claim to increase
the sexual potency of male and are aphrodisiac. So weplanned a project, the
aim of which was to study the metabolism of gold, evaluate its safety and
examine the claims about male sexual function (29).

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For this purpose, gold containing Ayurvedic preparation, Swarna Vasant
Malti, prepared by Director,NationalInstitute of Ayurved, Jaipur,was given to 20
male persons in a dose of 100 mg thrice daily for 40 days under supervision of
Ayurvedic physicians. This amounted to total cumulative intake of 160 mg of
gold at the rate of 4 mg per day. In this form, the gold preparation did not have
any toxic effect on human body as evidenced by clinical examination, unaltered
body weight, absenceof urinary pathology and by 30 sensitivebiochemical and
enzymatic tests. The gold fromthis Ayurvedic preparation was found in plasma
and erythrocytes, excreted partly in urine and was presentin semen. Gold
binding to albumin and hemoglobin slightly increased their electrophoretic
mobility towards anode. This finding was similar to whatwe had earlier observed
with mercury (19,22). Itappears thatmetal binding to these proteins alters the
net electric chargeon the molecule resulting into altered electrophoretic mobility
(29) which returns to normalwhen the metal is cleared fromthe blood (19,22).
More significantly, the study showed that gold was detected in semen
which increased significantly on therapy. Itwas also found that, on an average,
the volumeof ejaculate increased by 8%, spermcount by 6% , sperm motility by
17% and acid phosphataseby huge 73% (29). Westrongly recommend repetition
of this and similar work in large number of people for the benefit of infertile or
less fertile males.
In Ayurved , gold is not only used as bhasma , but foil as well (1), but no
reportis available on its effect. Therefore, the author himself offered to
volunteer for this study (30). Ten millgram of pure gold foil was taken by mouth
for 16 days with mawa sweet (milk cake). Routine blood chemistry was done
before and after gold therapy. All the blood values (chemical as well as enzymatic)
were well within normal rangeand variations, except the enzymes. The creatine
phosphokinase(CPK) and lactate dehydrogenase (LDH) decreased substantially
(as much as 25%) on gold foil ingestion while GOTand CPK-MB decreased about
12%. This indicates possible inhibition of these enzymes by ingested gold, but
whether this inhibition occurred in the body or during analytical procedureis not
clear. Alternatively the ingested gold may have caused decreased turnover of
heart tissue leading to reduced release of heart-specific enzymes in the

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circulation. If this possibility is substantiated , it may open a new area of research
for protection of heart. I fervently appeal to researchers to take up this project
for further studies.
COPPER: Copper is an essential trace element, has important role in iron
metabolism and hemoglobin synthesis (16,31). In Ayurved, copper bhasma and
various preparations containing it are widely used. So experiments, similar to
those using silver bhasma(24), wereconducted on chicks to know the effect of
copper bhasma on essential trace elements (31). Theingested copper bhasma
notably affected iron , significantly increasing it in plasma, whole blood and liver
of experimantal group chicks.
In another study, copper bhasma supplementation in ration of chicks
exhibited decrease in total lipids, phospholipids, triglycerides, totalcholesterol
but increase in HDL- cholesterol (32). Theseresults are surprisingly similar to what
we had observed with not only silver bhasma but other preparations of silver as
well (26). This similarity of biological effects reinforces the contention that
elements of same periodic group exhibit similar chemical as well as biological
properties. Perhaps, it is the wisdomof our ancient people that they advocated
drinking of water kept in copper and silver utensils in order to keep the blood
lipids in check.
STRONTIUM: The pearly white 32 sparkling teeth are everybody’s dream.
But this dream is marred by dental caries which is one of the commonest
affliction encountered by Dentists.
We know that calcium and strontium are members of sameperiodic group
2a (Fig.1) , so they sharesimilar properties. Strontium being heavier element can
displace calcium . While teaching in dental college, I had an occasion to do
research on dental caries.
First, we analyzed carious and non-carious teeth for 35 inorganic elements
by inductively coupled plasma (ICP) techniqueand found that strontiumwas the
only element found to be presentin significantly lower amountin carious teeth,
thus strongly suggesting that its deficiency may help formcaries (33).

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In second study (34), wholeblood of people having multiple caries versus
sound teeth was analyzed for these 35 elements to determine any possible
relation between chemical composition of a person’s blood to formation of caries.
Thirteen elements were not detected in blood by this technique. Of remaining 22
elements, the result of only seven elements (Fe,P,B,V,Sr,Sn, and F) were
significantly different between the two groups. Themost remarkablefinding of
this study was significantly decreased amount of phosphorus, strontiumand
fluorine and perhaps increased boron in the blood of persons with caries (34).
In the third study (35), the rate of calcium release fromextracted carious as
well as non-carious teeth on immersion in 1% lactic acid was determined before
after strontiumchloride treatment for one month. Itwas found that (i) The rate of
calcium release from non- carious teeth was significantly higher than carious
teeth, possibly because there was more calcium present, (ii) The rate of calcium
release was almost halved after strontium treatment in both groups of teeth, (iii)
the Vicker’s micro – hardness of non- carious teeth was higher than those of
carious teeth. Thus strontiumtreatment may be beneficial in reducing loss of
calcium fromintact teeth__ noncarious as well as carious (35).
The readers will appreciate that there is great scopeof quality research
work in dental biochemistry, even in relation to strontium. The interested readers
are referred to a recent review on strontiumand caries (36).
CALCIUM: Renal stone formation is very common in North India . These stones
mainly consistof calcium oxalate indicating abnormalmetabolism of calcium or
oxalate or both. Since large doses of ascorbic acid are advised for various
indications , and it forms oxalate in the body , it was decided to see if long
continued ingestion of large doses of ascorbic acid induces calcium oxalate
deposition in rats (37).
Itwas found that neither stone nor calcium oxalate crystaldeposition was
observed in the kidneys of rats after ingestion of 60 mg L-ascorbic acid daily for
three months , although urinary excretion of stone inhibitors (magnesiumand
citrate) was decreased and oxalate increased . The highly acidic pH of urine and
reduced calcium excretion might haveprevented their deposition (37).

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LEAD : Environmentalpollution of lead has been and is still a serious public health
hazard. Itoccurs through mining, smelting and refining of its ore galena (lead
sulfide) , burning of coal and petroleum fuels containing lead additives (38). A
common man is exposed to alkyl-lead compounds presentin motor exhausts.
Hence traffic policeman are expected to be worstaffected. Therefore, we had
undertaken study of blood lead concentration of traffic policemen as well as
activity of enzyme 5-aminolevulinate dehydratase( which is inhibited by lead)
and compareit with people from much less exposed areas (39). Theresults
showed that average lead content was one-and-half times in exposed group
(traffic policemen) and average AL AD was less than two-third of non-exposed
persons. Itconfirmed that ALAD activity is a good marker of lead exposure__
much easier to determine than blood lead (39).
SELENIUM: Selenium is placed in group 6a justbelow sulphur (Fig. 1) so its
biological chemistry resembles that of sulphur (38). Our interestin selenium arose
fromreported protective effect of selenium against mercury toxicity. Therefore ,
we conducted experiments using goldfish kept in aquarium to see the effect of
sodiumselenite and L-selenomethionine on the toxicity of mercuric chloride and
methylmercuric chloride . Our results showed that these selenium compounds
were not protective againstthese two mercury compounds (40).
Itis pertinent to mention that high selenium intake by cattle as well as
humans has been found to cause brittleness of the hair and hoofs in the former
and loss of hairs and nails in the humans. This has been explained on the basis of
selenium forming its compounds selenocystineand selenomethionine , analogous
to sulphur containing cystine and methionine. As hairs and nails are rich in these
amino acids , their replacement by selenium containing amino acids may cause
loss of hairs and nails (40). This could be a challenging but very rewarding
research projectas hair loss is a very common problemafflicting millions of men
and women.
NEWER ANALYTICAL METHODS

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In true spirit of laboratory biochemist, I havedeveloped newer analytical
methods of estimating certain inorganic elements (metals). These are mentioned
below.
(i) A new colorimetric method for determination of serum iron using
dimethylglyoxime (41).
(ii) A new atomic absorption spectrophotometric method for direct
determination of mercury in blood (without any pretreatment ) using
borohydridereduction (42).
(iii) A new turbidimetric method for determining urinary calcium(43).
MERCURY ONTHE CHEEKS , NICKEL ONTHE LIPS
Lastly, let me discuss two research areas which will definitely attract young
Biochemists. These belong to cosmetic industry which is a billion dollar industry in
the world.
Indians areobsessed with ‘fair skin’ and this obsession has reached to the
level of madness, especially in girls, with the result that several ‘fairness creams’
are available in the market . These are used with or without doctor’s advice. It
may be noted that “fairness creams” essentially contain mercury, a toxic element.
A survey conducted by “Center for Science and Environment“ in India (44) found
severalharmfulmetals (mercury, nickel, cadmium, chromium etc. ) in mostof the
creams , even of international brands. Sometimes their level approached the
dangerous levels, causing allergy and permanent scar.
Another dangerous cosmetic is “lipstick” which is used by almost every
woman, whether rich or poor. Its useby women stems fromtheir innate desire to
make their lips attractive, colored and juicy, withoutrealizing that they are
actually making them poisonous for themselves as well as their partner because
lipsticks not only contain nickel, but several other toxic metals (chromium,
aluminum, cadmium, manganese, lead, etc.) . These metals are absorbed by oral
mucosa and also swallowed with saliva thus ultimately proving harmful to the

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person using it. For example, cadmium may causecancer while lead is a
neurotoxin . The level of lead will be more in lipsticks of dark shades (44).
REFERENCES
1. Nadkarni AK . Dr. K.M.Nadkarni’s Indian Materia Medica, Vol.2, 1986,
Popuar Prakashan, Bombay .
2. Chopra RN, Chopra IC, Handa KL, Kapur LD. Chopra’s Indigenous
Drugs of India, 2nd ed., 1958 , UN Dhur & Sons Pvt. Ltd., Calcutta.
3. Sharma PV . Siddha Medicine. In ‘History of Medicine in India, (Ed.) PV
Sharma, The Indian National Science Academy, New Delhi,1992,
pp.445-450.
4. Boyd H. Introduction to Homeopathic Medicine. 1981, Beaconsfield
Publishers Ltd. , Beaconsfield.
5. Sharma, DC. Analysis of trace elements. Ind. J. Clin.Biochem.,1988, 3,
Spl. Suppl., 128-134.
6. Varian Techtron . Basic Atomic Absorption Spectroscopy, 1975, Varian
Techtron, Springvale.
7. Thiers RE . Contamination in Trace Element Analysis and its Control. In “
Methods of Biochemical Analysis”, Vol. 15, ( Ed.) D Glick, Interscience,
New York,1957, pp. 273-335.
8. Gorsuch, TT. The Destruction of Organic Matter. 1971. Pergamon
Press, Oxford.
9. Schwarz K, Milne DB, Vinuyard E (1970) . Growth effects of tin
compounds in rats maintained in a trace element – controlled
environment. Biochem. Biophys. Res. Commun. 40, 22-29.
10. Schroeder HA. The Trace Elements and Man: Some Positive and
Negative Aspects . 1973. Elsvier, Amsterdam.

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11. Underwood EJ . Trace Elements in Human and Animal Nutrition , Vol.
1 and 2 , 4th
ed., 1977, Academic Press, New York.
12. Mertz W (ed.). Trace Elements in Human and Animal Nutrition, 5th
ed.
1989 , Academic Press , New York .
13. Gray T . The Elements. 2012. TheBlack Dog & Leventhal Publishers,
New York .
14.Sharma DC . In vivo relationship of iron with certain metals and the
effect of iron deficiency on enzymes. Ph.D. Thesis . 1972. University of
Udaipur , Udaipur.
15.Sharma DC, Sharma M, Rathore AS, Gupta OP , Dube MK , Simlot MM
(1980). Effectof silver, gold and mercury colloids on erythrocyteand
iron metabolism. Ind. J.Exptl.Biol. 18, 1309-1311.
16.Sharma DC, Mathur R , Singh PP (1993). Iron metabolism: A review ,
Ind. J.Clin. Biochem. 8, 80-101.
17. Sharma DC ,Mathur R (1995) Correction of anemia and iron deficiency
in vegetarians by administration of ascorbic acid. Ind. J. Physiol.
Pharmacol. 39, 403-406.
18. Sharma DC ,Chandiramani D, Riyat M , Sharma P . (2007). Scientific
evaluation of someAyurvedic preparations for correction of iron
deficiency and anemia. Ind. J. Clin. Biochem. 22, 123-128.
19. Sharma DC . Biochemical studies on someaspects of toxicity and
metabolism of mercurials. Ph. D. Thesis, 1982, University of Rajasthan
, Jaipur .
20.Sharma DC (1987). Biochemicalbasis of the toxicity of mercury. Med.
Hypotheses, 23, 259-263.
21.Sharma DC, Davis PS , Sharma PK. (1981). Studies in search of
modifiers of the toxicity of mercurials and speculations on its
biochemical mechanism. Biochem. Pharmacol. 30, 3105-3107.
22.Sharma DC, Sharma PK , Singh PP. (1986). Detection of mercury
poisoning by altered electrophoretic mobility of blood proteins . Clin.
Chem. 32, 1595-96.

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23.Sharma DC , Jha J , Sharma P, Gaur BL (2001). Evaluation of safety and
efficacy of a gold containing Ayurvedic drug. Ind. J. Exp. Biol. 39, 892-
896.
24.Sharma DC , Dadheech G , Fiza B, Mathur M, Riyat M ,Sharma P. Effect
of oral ingestion of different forms of silver on tissuecontent of some
essential elements in chicks. Ind. J. Clin. Biochem.2009 : 24, 202-204.
25.Hirasawa F, Sato M, Takizawa Y. (1994). Organ distribution of silver
and the effect of silver on copper status in rats. Toxicol. Lett. 70,193-
201.
26.Sharma DC ,Budania R , Shah M, Jain P, Gaur BL (2004). Hypolipidemic
activtity of silver preparations in chicks, Gallus serregineus. Ind. J.Exptl.
Biol. 42,504-507.
27.Sharma DC , Sharma P, Sharma S (1997). Effect of silver leaf on
circulating lipids and cardiac and hepatic enzymes. Ind.J.Physiol.
Pharmacol. 41,285-288.
28.Jha J . A biochemical study of toxicity and metabolism of gold. PhD.
Thesis.1999, University of Rajasthan, Jaipur.
29. Sharma DC, Jha J, Sharma P , Gaur BL (2001). Evaluation of safety and
efficacy of a gold containing Ayurvedic drug. Ind.J.Exp.Biol. 39,892-896.
30.Sharma DC , Sharma P (2013). A preliminary study of the effect of
ingestion of gold on blood chemistry. Ind.J. Clin. Biochem. 28,303-304.
31. Sharma DC, Kochar B, Bhardwaj, A , Riyat M, Sharma P (2009). Effect
of ingestion of copper bhasmon red cell indices, iron parameters and
essential elements in chicks. Ind. J. Clin. Biochem. 24,245-249.
32.Choudhary P ,Chadda A Kochar B , Sharma DC (2007). Effectof copper
bhasma (an Ayurvedic preparation ) supplementation on circulating
lipids of chicks. Ind.J.AnimalSci. 77, 566-568.
33.Riyat M , Sharma DC(2009). Analysis of 35 inorganic elements in teeth
in relation to caries formation . Biol. Trace Elem. Res. 129, 126-129.
34.Riyat M , Sharma DC (2010). Significanceof trace element profile of
blood of persons with multiple caries versus sound teeth . Biol. Trace
Elem. Res. 134,174-179.

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35.Riyat M , Sharma DC(2010). An experimental study of the effect of
strontiumpre-treatment on calcium release fromcarious and non-
carious teeth. Biol. Trace Elem. Res. 133,251-254.
36.Lippert F, Hara AT (2013). Strontiumand caries : A long complicated
relationship. Caries Res. 47, 34-49.
37. Singh PP, Sharma DC, Rathore V, Surana SS (1988). An investigation
into the role of ascorbic acid in renal calculogenesis in albino rats. J Urol
139, 156-157.
38. Sharma DC (1995). Essentiality and toxicity of certain toxic elements : A
Strategy, In “ EnvironmentalStrategies “, (Ed.) RM Lodha , Himanshu
Publications, Udaipur, pp. 17-27.
39.Sharma DC, Seervi N, Rawtani J (2000). Effectof environmental lead
pollution on hemoglobin and erythrocyteALAD activity. Ind J Physiol
Pharmacol 44, 117-118.
40. Sharma DC , Davis PS (1980). Effectof sodiumselenite and
selenomethionine on the accumulation and acute toxicity of mercuric
and methylmercuric chloride in the goldfish (Carassiusauratus). Ind J
Exptl. Biol. 18, 82-84.
41. Sharma DC , Singh PP, Khalsa JK (1969). Determination of serumiron
with dimethylglyoxime. Clin.Biochem. 2,439-445.
42. Sharma DC, Davis PS (1979). Directdetermination of mercury in blood
by use of sodiumborohydridereduction and atomic absorption
spectrophotometry. Clin.Chem. 25,769-772.
43. Sharma DC, Rathore V , Kiran R, Singh PP (1987). A simple and rapid
turbidimetric method for determining urinary calcium. Ind. J. Exp. Biol.
25,855-858.
44.Center for Science and Environment. Presenceof heavy metals in
cosmetics. 2014. A CSE Study Press Release, New Delhi , January 15,
2014.

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