Chapter V Discussion

Among the environmental factors those may contribute to the genesis of human cancer, diet is considered as a major factor. Experimental and epidemiological studies have shown that diet makes a substantial contribution to the increasing burden of human cancer. Various epidemiological studies indicate that the consumption of cooked meat and meat product predisposes individuals to higher risk of cancer, particularly, colon, rectal, stomach and Breast cancer (Doll, 1992; Walters et al., 2004). Dietary factors which may be important in the aetiology of human cancer include heterocyclic amines (HAs), which are formed during cooking of red meat and other non-vegetarian food. A series of heterocyclic amines (HAs) have been isolated as mutagens and carcinogens from various kinds of heated materials including cooked meat and fish, and pyrolysis products of amino acids and proteins. These mutagenic HAs are classified into 2-amino-3methylimidazo[4,5-f]quinoline IQ.-type HAs, having a 2-aminoimidazole moiety as a common structure, and non-IQ-type compounds, having a 2-aminopyridine moiety as a common structure. Among them 10 HAs have been demonstrated to be carcinogenic in rats and mice. Quantification of HCAs in cooked foods and human urine samples has provided unequivocal evidence that humans consuming non-vegetarian food are continuously exposed to HAs in daily life. In fact, 2-amino-3,8-dimethylimidazow4,5-f xquinoxaline MeIQx.-DNA adducts have been found in DNA from human tissues, such as colon, rectum and kidney. Several epidemiological studies are now available, indicating a positive association between consumption of cooked meat and development of colorectal, pancreatic and urothelial cancers. In addition, it has been demonstrated that the highest risk genetically determined phenotype rapid CYP1A2 and rapid NAT-2., combined with a dietary preference for well-done red meat, is associated with a relative odds ratio of 6.45 for colorectal cancer risk. These findings suggest

that even if human exposure to HAs is at microgram levels per day per person, DNA adducts may be formed in human tissues and may significantly contribute to human cancer. Therefore, it is advisable that exposure to HAs should be decreased as far as realistically possible. Several measures reduce the extent of exposure to HAs. They include: 1. wrapping meat and fish in aluminum foil to avoid direct contact with a naked flame; 2. preheating of meat to remove creatinine in a microwave oven; 3. removal of charred parts of grilled meat and fish with a knife and fork, or chopsticks. In addition, supplement with antioxidants and antimutagens is considered to be effective, because free radicals are thought to be involved in HA formation through the Maillard reaction. In fact, the addition of butylhydroxyanisole BHA. to beef reduces its mutagenicity after frying. Mutagenic and carcinogenic MeIQx and 2-amino1methyl-6-phenylimidazow4,5-bxpyridine, PhIP are relatively abundant HAs in cooked foods, their precursors being creatinine, amino acids and sugars in meat and fish. MeIQx and PhIP can be formed by heating mixtures of creatinine, glycine and glucose, and of creatinine, Lphenylalanine and glucose, respectively. As part of a search for effective methods to decrease formation of HAs, the influence of various antioxidants, either food components or food additives, on formation of MeIQx and PhIP were examined by several researcher. Green tea catechins, epigallocatechin gallate (EGCG), quercetin, luteolin and caffeic acid proved capable of markedly reducing the levels of these HAs produced by mixtures of creatine, amino acids and sugars. Possible application of these antioxidants which are abundantly available in variety of spices, in cooking can play as preventive measure in formation HAs or inhibition of formed HAs, thus reducing possible risk of cancer. It is believed that Spices is the reason that brought Romans, Jews and Arabs to India. The search for spices was also the impetus for Christopher Columbuss discovery of America and for Vasco de Gamas voyage from Portugal to India, in the 15th century, along what is now called the Spice Route. The Indonesian island where the nutmeg, cloves, cinnamon, ginger,

turmeric and mace were grown is now called Spice Island. From ancient times, spices have played a major role in the lifestyle of people from certain parts of the world. They have served numerous roles through history, including as coloring agents, flavoring agents, preservatives, food additives and medicine. The active phytochemicals derived from these spices have provided the molecular basis for these actions. A spice is a dried seed, fruit, root, bark or flower of a plant or a herb used in small quantities for flavor, color or as a preservative. Many of these substances are also used in traditional medicines. Globalization has made these spices easily available, and increasing their popularity.

Indian food are most commonly cooked along with variety of spices as a mixture (blend) of spices popularly called as Garam Masala. Eleven different dried spices were blend to prepare Garam Masala, containing Black pepper with 40% and nutmeg with 0.4%. These spices known to contains various phytochemicals which posses medicinal potentials as well as antioxidants, antimutagens and anticarcinogens. Sinces, non-vegetarian foods (meat and fish etc.) are cooked with rich of spices, Garam Masala, it was of our interest to find out the possible antimutagenic/inhibitory/modulatory effects on the mutagenicity induced by 9 different HAs (IQ, MeIQ, MeIQx, MeAC, Glu-p-1, Glu-p-2, PhIP, Trp-p-1 and Trp-p-2) in TA98 tester strain of Salmonella typhimuriums in the presence of S9 mix. Garam Masala Ethanol Extract (GMEE) first evaluated for possible mutagenic potential and found non-mutagenic in all 5 tester strains, TA97a, TA98, TA100, TA102 and TA104 of Salmonella typhimurium both in the presence and absence of metabolic activation. Results revelas clearly that GMEE do not possess any phytochemical that can induced mutagenic potential in an in vitro Ames test. However, GMEE shows clear modulatory effects when it was evaluated against 9 different HAs. This modulatory effects of GMEE was observed as antimutagenic (protective) as well as co-mutagenic against certain HAs. GMEE at different

doses was found to be clearly antimutagenic against only one HAs, PhIP. GMEE at lower doses found to be co-mutagenic by enhancing the original mutagenic activity of 4 HAs, IQ, MeIQx, Glu-p-1 and weak co-mutagenic effects against Glu-p-2. GMEE at higher doses, 1 and 2 mg/plate could able to inhibit the mutagenic response of these 4 HAs, showing clear antimutagebnic effect. GMEE did not show any anitumtagenic effects against 4 HAs, MeIQ. MeAC, Trp-p-1 and Trp-p-2, rather found to be comutagenic enhancing the mutagenic potential of all these HAs than their original mutagenic activity. It is reported that some the agents are know to play the role of aantimutagen, mutagenc and co-mutagens at certain conditions against ceratin chemical mutagens and carcinogens, such agents are also called as Janus agents (Holm et al., 1988). GMEE shows similar kind of effects as antimutagn by inhibiting the mutagenicity of certain HAs and co-mutagen by enhancing the mutagenic activity of certain mutagenic HAs. All HAs require exogenous metabolic activation, phase I cytochrome P450 enzymes (S9 mix.) in the in vitro assays like Ames test to show their mutagenic response. Has are mainly metabolized first by cytochrome P450 (CYP) 1A2 in rodents as well humans. Other P450 isozymes including CYP1A1, 1B1 and 3A4 are also able to to some extent, for oxidation of the exocyclic primary amino group to a hydroxyamino group (Sugimura et al., 2004). These HAs are pro-mutagns and are converted by CYP microsomal enzymes in the form of active mutagenic and carcinogenic metabolites. Many phenolic compounds are known to inhibits the activity of microsomal cyp activity blocking the metabolic conversion of HAs and other activation dependent carcdinogens (Kassie et al., 2003; Schwab et al., 2000; Shishu and Kaur, 2002; 2008). Schwab et al (2000) evaluated the database on the dietary constituents that protect against DNA damage and cancer induction by heterocyclic amines (HAs). These compounds are well described as cooked food mutagens and carcinogens are formed from meat and fish upon prolonged cooking at high temperature. More than 80% of the antimutagenicity studies

against HAs are carried out with an in vitro Ames Salmonella/microsome mutagenicity test (Schwab et al., 2000). In these experiments, exogenous liver homogenate containing phase I enzymes is added in order to reflect the bioconversion of HAs to DNA reactive electrofiles in vivo. However the activation capacity of the S9 mix can be modified by different factors like changes in molarity and pH value of the S9 mix caused by a putative antimutagns reduce the activation capacity of the enzyme mix. However, critical survey was that conventional in vitro tests used in many of the studies do not fully reflect the complex activation and detoxification processs which take place in mammals. Several dietary constituents which were DNA protective under the in vitro conditions in experiments with indicator cell which require addition of exogenous activation mixture were inactive in in vivo rodent assays or even led to enhancement of the DNA damaging properties of HAs. But certain conventional in vivo experiments with laboratory animals are not found to be adequate tools to identify protective constituents in human foods since only marginal or negative effects are obtained in these models with HAs (IARC, 1993; Kassie et al., 2003). In the present investigation GMEE could able to modulate the mutagenic activity of all 9 mutagenic/carcinogenic HAs either by inhibiting or promoting the mutagenic activity of certain HAs. These HAs are generated during cooking of non-vegetarian food, meat and fish and their quantity of formation is depend on several condition like type of meat, cooking time and temperature (Sugimura et al., 1977, Sugimura and Sato, 1983; Surh, 1999; Spingarn et al., 1980). HAs are formed at nano or miro-gram level during cooking, but these HAs are strongly mutagenic at these level. Some of most commonly formed HAs, like MeIQ, PhIP, IQ etc are 100 and 1000 fold more mutagenic than very well known mutagens and carcinogens, benzo(a)pyrene and aflatoxin B1, respectively (Sugimura et al., 1993).

Generally, in most of the antimutagenesis studies against certain known mutagens and carcinogens, one of the optimum dose of mutagens is selected with different doses of test agent/possible antimutagens, of interest. Initially, we have used single optimum dose of HAs with 4 different doses of GMEE. Later 3 diffrent doses of HAs as low, medium and high are used with 4-5 doses of GMEE, to study the antimutagenic and co-mutagenic response. Interestingly, comutagenic effects of GMEE was very strong at low doses of HAs, like Glu-p-1, MeIQ, MeAC, Trp-p-1 and Trp-p-2. GMEE at 0.1 mg/plate could able to potentiate (promote) the mutagenic activity by 318%, 268% and 26% of MeIQ at the dose of 0.001, 0.01 and 0.1 g/plate, respectively (Tabloe 4.5). MeIQ at 0.1 g/plate induced very high number of revertant colonies, 6745 and GMEE at 0.01 mg enhanced revertant number to 8786 resulting 30% increase. MeIQ at median dose of 0.01 g/plate, revertant colonies were 1293 which is further enhanced to 5077 by GMEE at 0.5 mg/plate, resulting 292% increase. Whereas, MeIQ at low dose of 0.001 g/plate induced revertant colonies were 200 which was further enhanced to 837 by GMEE at 0.1 mg, resulting to 318% increase (Table 4.5). Similar co-mutagenic effects of GMEE was observed with other HAs, Glu-p-1 (Table 4.8), Trp-p-1 (Table 4.10), Trpp-2 (Table 4.11) and MeAC (Table4.12). Co-mutagenic effects of GMEE was observed at high extent at low and median doses of HAs. Issue of co-mutagenicity with Has has been also reported by Sugimura et al. (2004). During purification of Has from tryptophan pyrolysate, a sudden loss of mutagenicity was observed in certain fractionation steps. Remixing of the fractions restored the original mutagenic activity at accelerated extent with strong comutagenicity. Mutagenic HAs having quinoxaline, indole and methyl methylpyridol imidazole moity in MeIQx, MeAC, Trp-p-1, Trp-p-2 and Glu-p-1 are found to enhance their mutagenic potential at low doses by GMEE in the presence of CYP in rat liver S9 mix. Mutagenic activity of HAs having pyridine and imidazole moity in PhIP and Glu-p-2 are only inhibited by GMEE at all low and high doses, used in this investigation.

Fifty years of research on carcinogenesis have identified multiple cancer risk factors in the human environment, including food. Attempts to counteract carcinogenesis have led to discoveries of many components of plant foods that display biochemical and biological activities capable of preventing cancer. Anticarcinogenic food components have become the subject of interest of a relatively new cancer research field called cancer chemoprevention. Although many data on carcinogenic and anticarcinogenic food components have been collected, our understanding of causal mechanisms linking diet and cancer is still evolving.

The link between diet and cancer is rather complex and definitely not unidirectional. The majority of food products are neutral in relation to carcinogenesis. Only some exhibit proor anticarcinogenic properties. Obviously, an abundance of procarcinogenic food constituents is as harmful as a lack of anticarcinogens in the diet. The point, however, is that such a categorization is easily applicable only to a single chemical compound, while dietary products are almost exclusively complex mixtures. Hence, there are numerous situations when a single food product contains, at the same time, some substances that are harmful and some that are beneficial. Garam Masala shows similar kind of effects against HAs evaluated in this investigation. GMEE shows both antimutagenic as beneficial and comutagenic as harmfull effect, this is dose dependent as found in this in vitro investigation.

Garam Masala used in this investigation contains 11 different spice material at different proportion, like black pepper (40%), black cardamom, caraway, clove, cinnamon (8%), bay leaf, stone flower, mesua ferrea, Mace, Green cardamom, and nutmeg (0.4%, Table 3.1). Each of spices contains several phytochemicals which are reported as antioxidant, antimutagenic and anticarcinogenic against certain chemical mutagens and carcinogens. As GMEE shows both antimutagenic as well as co-mutagenic effects against certain HAs, which of the GM spice is responsible became our interest. Since black pepper used as major spice ingredient (40%) of

Garam masala, extract was prepared and along with pure piperine also evaluated for their possible antimutagenic and co-mutagenic activity against 3 HAs, IQ, MeIQ and PhIP. Black pepper ethanol extract (BPEE) and piperine was also found co-mutagenic promoting the mutagenic activity of MeIQ (0.01 g/plate) by 460% and 520%, respectively (Table 4.14). GMEE at similar dose of MeIQ promoted mutagenic activity by 268% (Table 4.5). BPEE and piperine found to have higher co-mutagenic activity compared to GMEE. Compare to GMEE, BPEE and piperine also shows stronger co-mutagenic effects against another HA, IQ (Table 4.6 and 4.16). However, like GMEE, BPEE also exhibited antimutagenic activity against PhIP (Table 4.7 and 4.15). Pure phytochenical of black pepper, piperine did not exhibited antimutagenic activity agsint PhIP rather found weak co-mutagenic activity. By increasing the bioactivity of other anti-carcinogenic spices, black pepper dramatically increase their potency and effectiveness against cancer. In addition to this, black pepper also counteracts cancer development directly. Its principle phytochemical, piperene inhibits some of the pro-inflammatory cytokines that are produced by tumour cells. In so doing it interferes with the signalling mechanism between cancer cells, there by reducing the chances of tumour progression. Collectively, this property makes black pepper one of the most important spices for preventing cancer.

Since Garama Masla Extract and Black pepper as major component of GM shows almost similar kind or slightly higher co-mutagenic activity against IQ and MeIQ and antimutagenic activity against PhIP, fresh GMEE was prepared with and without black pepper evaluated for antimutagenic and co-mutagenic activity against 3 HAs, MeIQ, IQ and PhIP. GMEE with and without black pepper as well as BPEE shows similar level of co-mutagenic effects against MeIQ (Table 4.17). GMEE with and without black pepper shows co-mutagenic

at two lower doses and antimutagenic at two higher doses and clear antimutagebic effects against PhIP (Table 4.18). Results of the present finding shows that Garam masala with and without black pepper shows both antimutagenic as beneficial and co-mutagenic as adverse effects against certain carcinogenic HAs. In our cooking style when meat and fish are cooked with garama masala or other kind of spices, it may help in inhibiting the mutagenic risk of certain HAs and it could also might play harmful role by promoting mutagenic and carcinogenic activity of certain HAs. It is well knows from several findings that certain plant extract or phytochemical found antimutagenic and anticarcinogenic against certain mutagen/carcinogen, same may show no antimutagenic effects as well as co-mutagenic effect against different clas of chemical mutagen/carcinogen (Furguson, 1994; Surh, 1999). With this view, GMEE was evaluated for its antimutagenic and co-mutagenic activity against 6 different activation dependent mutagens/carcinogens, Aflatoxin B1 (AFB1), Benzo(a)pyrene (BaP), 7,12-dimethylbenz anthracene (DMBA), cyclophosphamide (CP), 2-aminofluorene (2-AF) and 20-

methylcholanthrene (MC) in TA100 tester strain in the presence of S9 fraction. Except, 2-AF, GMEE found to have very good antimutagenic effects against all 5 mutagens and carcinogens (Table 4.22 and 2.23; Fig. 4.16). N-acelation is an essential step in metabolism of 2-AF, and there is possibility of GMEE failing to inhibit the N-acylation resulted in negative antimutagebic effects against this mutagens. No comutagenic effects of GMEE was found against all 6 mutagens at both lower and higher dose, unlike effects observed with HAs. HAs like many carcinogens and mutagens, Aflatoxin B1, benzo(a)pyrene etc. require metabolic activation in order to extert its mutagenicity and this is highly dependent upon cytochrome P450 (CYP) 1A2, 1A1. Further metabolism is through O-acetylation, catalysed by N-acetyltranferase (NAT), but the N-acetoxy ester derivative form is unstable and spontaneously converted to a reactive epectrophilic arynitronium ion. This ultimate metabolite

is able to bind covalently to DNA and form adducts that may cause mutation and lead to induction of cancer. Most of the plant phenoclic and polyphenolic phytochemicals shows antimutagenic and anticarcinogenic effects by inhibiting the cytochrome P450 (CYP) isozymes or by blocking the metabolically activated mutagens and carcinogens. Antimutagenic activity of GMEE against certains HAs, PhIP, Glu-p-2, IQ, MeIQx and other carcinogens, AFB1, BaP, CP, DMBA and 20-MC might be due to inhibiting the Cytocrome P450 enzymes required for metabolic activation or by blocking the metabolically activated mutagenic species of these carcinogens in the in vitro assays like Ames test, used in the present investigation. Garama Masala ethanol extract (GMEE) shows very good antimuagenic effects against 2 HAs, PhIP, Glu-p-2 and other 5 carcinogens. Some of the phytochemical present in GMEE posses beneficial effects of antimutagenesis and might help in preventing the formation cancer in rodent as well as human at appropriate condition. However, GMEE has also ability to exhibit adverse effects by promoting the original mutagenic activity of few HAs. Use of Garam masala (spices blend) in cooking of vegetarian and non-vegetarian food might be helpful in inhibiting the mutation of certain carcinogens as well harmful in promoting the mutagenic activity of certain carcinogenic HAs. Human metabolic polymorphisms in the enzymes catalyzing the activation and/or detoxification pathways of carcinogen metabolism may account for differences in susceptibility to carcinogens between individuals. For example, CYP1A2 and N-acetyltransferase-2 (NAT2) are polymorphically distributed in humans and differences in the activities of these enzymes are postulated to be risk factors for cancer on exposure to heterocyclic amines (HAs). The rapid CYP1A2 phenotype combined with the rapid NAT2 phenotype was found to give an odd ratio of 2.79 for the risk of developing colorectal cancer in normal subjects and an 8.8-fold in smokers preferring well-done red meats (Le Marchand et al., 2001). In many western countries as well in modern Indian food style, the well-done meats and fish are some of the most favored

dishes. These foods are likely to produce large amounts of heterocyclic amines. These heterocyclic amines formed in fried, broiled or grilled meats and fish are metabolized to carcinogens by CYP1A2. CYP1A2 causes N-hydroxylation of the heterocycline amines which is followed by O-acetylation of NAT2 in the liver and then the intermediate moves to the target site to form nitrenium ion and then attacks DNA to initiate carcinogenesis. Therefore, an individual with a rapid phenotype for CYP1A2 will be susceptible to colorectal cancer in rapid NAT2 acetylators and to lung adenocarcinoma in slow NAT2 acetylators when enjoying welldone meats and fish. As GMEE found to have co-mutagenic activity agisnt few HAs, it might be due to the increasing the activity of microsomal CYP1A2 present is S9 mix by enhancing the N-hydroxylation of HAs, like, IQ, MeIQ, MeAC, Glu-p-1, Trp-p-1 and Trp-p-2. If this be the case also for in vivo condition, spices may also enhance the carcinogenic activity of HAs in animal and human exposed or consumed appropriate amount of well done meat and fish products. Antimutagenic effects of few pure phytochemicals, ellagic acid, chrlorophyllin, curcumin, carvacrol, eugenol, cinnamic acid, cinnamaldehyde and cinnamon oil was evaluated against two HAs, MeIQ and PhIP. Since GMEE shows co-mutagenic and antimutagenic activity against MeIQ and PhIP, respectively, possible antimutagenic activity of 7 known antimutagenic and anticarcinogenic pure phytochemicals were evaluated only against these two HAs. All these phytochemicals shows clear co-mutagenic activity against MeIQ. Even very well document curcumin used as coloring and flavouring agent of our daily food also shows comutagenic activity against MeIQ. Except ellagic acid and curcumin, all other 5 phytochemicals shows good antimutagenic activity aginst PhIP. Some of these phytochemicals, like carvacrol, eugenol, cinnamic acid cinnamaldehyde are present in some of the spice components, caraway, clove and cinnamon, respectively used in preparation of Garama Masala Extract.

Many researchers have discovered that like many spices clove contains antioxidants. Antioxidants help prevent the cell damage that scientists believe eventually causes cancer. On the other hand, in laboratory tests, the chemical eugenol, has been found to be a weak tumour promoter, making clove one of many healing herbs with both pro and anticancer effects. Preliminary studies have suggested the chemoprevention potential of clove for lung cancer and to reduce the formation of skin cancer. One dietary polyphenol which has been of focus of considerable attention is curcumin. It is the major constituent of spice turmeric, obtained from dried powdered rhizome of Curcuma longa (Aggarwal et al., 2007). The turmeric or its major yellow pigment curcumin (C) has been reported to possess antioxidant, anti-inflammatory inhibitory activity against chemically-induced carcinogenesis in several experimental models. Curcumin has also been shown to be a potent inhibitor of several environmental mutagens requiring metabolic activation (Nagabhushan, et al, 1987). Curcmin was also reported to be as antimutagenic against several HAs in an in vitro Ames Salmonella mutagenicity test (Shishu and Kaur, 2002 and 2008). However, our findings are contradictoary as curcumin found to be co-mutagenic against MeIQ and not antimutagenic against PhIP. Antimutagenic activity of any plant extract, phytochemical or pure synthetic chemnical found in the in vitro mutagenicity bioassays need to be evaluated using an in vivo rodent cytogenetic test like mouse bone marrow micronucleus test. When one of the HA, IQ was evaluated for cytogenetic activity, no clear statistically or biologically significant induction of micronuclei was observed in PCEs at any dose of IQ treated animals. All HAs shows very strong mutagenic activity in an in vitro Ames Salmonella/microsome mutagenicity test, but not much data available as positive mutagtenic response of HAs in the in vivo rodent mutagenicity tests (Sugimura et al., 2004). Since, failure to induce clear mutagenic effect in an in vivo mose bone marrow micronucleus test by IQ, antimutagenic effects could be taken up with GMEE.

Sasaki et al., (1997) evaluated mutagenicity of HAs, Trp-p-1, Trp-p-2, IQ, MeIQ, MeIQx and PhIP in different tissues of exposed mice. Some of the HAs induced DNA damage in mose liver, lung, kidney but failed to induced DNA damage in bone marrow (Sasaki et al., 1997). Present study also revela that when IQ exposed to mice, did not induced DNA damage in the form of micronucleated PCEs in mose bone marrow. Few chemicals like quinolone drugs are also reported to be strong mutagenic response in the in vitro test but failed to induce mutagenic response in the in vivo tests. Pattern of absorption, distribution and metabolism of chemical carcinogen in the rodent might be playing some role in failing to achive mutagenic response in the in vivo rodent bioassays. HAs are also reported to be strongly mutagenic in vitro Ames Salmonllea test but failed to induce mutation in the mouse bone marrow micronucleus test. HAs on the other hand reported to be carcinogenic in the long term rodent bioassay and also found to be associated in the development of human cancer risk in peoples consuming high amount of non-vegetarian food (Sugimura et al., 2004). It has been well documented that the use of herbs in Indian and Chinese medicine as a popular tradition for local remedies, have been practiced for the last 2000 years or longer. In actuality many of the present day drugs have evolved from meticulous Indian and Chinese herbal concoctions. The majority of these herbal therapies or Herbal Cancer Treatments are predominantly used, to rejuvenate and strengthen the body's natural immune system. These preparations are essentially in the form of complex and surreptitious formulas, essentially consisting of medicinal herbal cocktails and other ingredients, which are normally dried up and ground into pill or capsule form. These powerful herbal cancer treatments are also typically made in concoctions of liquid form, ideally containing two or more main ingredients to target specific maladies such as Cancer. Where some components are meant to act chiefly on the infirmity, while the others cancel out any harmful toxicity that may be present within the basic formula itself.

Regardless of these disappointing truths; at present scores of mainstream researches such as microbiologists, pharmacologists, botanists and chemists who endorse genuine natural remedies, are combing the entire earth's surface in search of herbs and plants with distinctive phytochemicals qualities. This is done with the greatest of enthusiasm in the hope that whatever new discoveries are made, it could possibly be developed to treat and cure diverse diseases. Moreover, many conventional campaigners clandestinely acknowledge that although Herbal Cancer Treatments usually take effect only over a period of time, it supplies crucial immunological and nutritional sustenance to the body which chemical-based pharmaceutical drugs are just unable to offer. Garam Masala prepared in this investigation contains 11 different spices, dried part of spice plants. Each of the spices contains phytochemicals many of them are reported to have antioxidants, antimutagenic and anticarcinogenic potentials against certain chemical mutagens and carcinogens. Results of the in vitro finding on the modulatory effects of Garam Masala and few pure phytochemicals clealy shows its antimutagenic as well as co-mutagenic activity against heterocyclic amines (HAs). Use of spices while cooking o meat and fish may some times help in the inhibiting the mutation of certain HAs as well as promoting the formation as well as potentiating the mutation induced by certain HAs. Similarly, few phytochemicals and particularly, caffeine was reported to be anticarcinogenic against PhIP induced rat mammary gland carcinogenesis, but acts as a co-carcinogen inducing the cancer rate in colon cancer (Takeshita et al., 2003). Therefore, further, long term cancer bioassays may be very helpful to know the anticarcinogenic and co-carcinogenic activity against HAs induced cancer in mice or rats.