Quantitative Estimation of Fungicide , Pesticide Residues from Pepper ( Piper nigrum L . ) as Obtained in the Selected Area of Kodagu District , Karnataka

Kodagu the one of the major pepper cultivated district in Karnataka state. The present study was carried out in the different villages such as Tuchumaleri, Chikkamandur, Mattur, Nallur, Kunda, Balaji, Kottur, and Mallanakere etc of Ponnempet, Kodagu district. During the pepper cultivation the farmers are used the pesticides in each steps, for high yielding purpose. This study was revealed that to measure the amount of pesticides present in the final product. Aflatoxin is produced by the fungi aspergillusflavus and it having properties of nephrotoxic, teratogenic, carcinogenic, and immunosuppressive. It can be determined by the reliable method like HPLC (column symmetry C-18; 4.6X250 mm) used with detection by florescence (excitation 364 nm, emission 465 nm). Like that the organo phosphors also determined by GC (column summery RTX-5; 30m) the level can be indicated in ppb and ppm respectively.


INTRODUCTION
Pepper (Piper nigrum L.) belongs to the family Piperaceae, the crop grow well in a temperature range between 25°-40°C and rainfall around 1250 mm.In India, Karnataka, Kerala, Tamilnadu states are play a major role in the production of pepper.As per the 2013-14 report 21,250 tones of black pepper worth Rs.94,002 lakhs products were exported from India to various countries.
Pepper is one of the important spice used for flavor, and aroma of food and beverages, is produced from the still-green, unripe drupes of the pepper plant.Pepper contains volatile oil so it makes an important contribution to sensory qualities 1 In Indian ayurveda system the pepper are used as a medicine since 5000 years, as a diet and herbal remedies, it is a source of important drugs such as atropine, codeine, digoxin, morphine, quinine and vincristine 2 .While emphasizing the body, mind and spirit in disease prevention and treatment 3 .
Piperine has been found to enhance the therapeutic efficacy of many drugs, vaccines and nutrients by increasing oral bioavailability by inhibiting various metabolizing enzymes 10 .The consumption of black pepper increases the orocecal transit time. 11and also decreased pressure in arteries, it inhibited high K + pre contractions due to blockade Ca 2+ channel 12 , apart from that it having an anti diarrheal property hence in most of the developing countries local peoples, herbal industries, and herbal practioners formulate the peppercorn for diarrhea for all ages 13 .
Nerolidol is a natural compound isolated from piper species, having pesticidal activity against various mites.And another compound pipene isolated from volatile oil of peppercorn it is used as an odorant 14 .

Collection of sample
Kodagu is one of the major pepper cultivated district in Karnataka state hence the pepper (Piper nigrum L) samples are collected from the different villages of ponnempet.The samples were collected in a zip polythene bags.And preserved in deep freezer.

Survey in farming practices
Questionnaires contained four sections to find out the details like Physical inspection (location, soil type and weather data) general information (type of crop, aware of pesticide effect and recommended dosage) cultivation steps (name of the verity, land preparation, spacing, month of plantation, intercropping, duration and common diseases) pesticide information (name of pesticide used, method of application etc.)

Stock and working standard
Stock and working standard of organo phosphorous pesticides of appropriate concentration in acetone (standard from EPA of equivalent).Store the standards under refrigeration.(Shelf life for stock standard is one year and that for working standard is six months) Florosil Cleanup for Organo phosphorous • Place activated florosil (about 4") in the column; add about 0.5" sodium sulphate pre wet column with 50 ml petroleum ether.

Fig. 2: Aflatoxin standard
• Transfer the concentrate to sintered column using 8ml HPLC grade acetone, and rinse walls of chromatographic tube with additional small portion of acetone.Drain with 100 ml of petroleum ether and collect and collect it in a 500 ml beaker.Make up to 4ml with acetone.This is ready for GC analysis 16 .

RESULTS AND DISCUSSION
Kodagu, one of the major pepper cultivated district of the state Karnataka.A total of ten farmers having plantations with pepper crop from different villages of ponnempet, kodagu district were interviewed.They were majorly cultivating pepper plants.Distance of 10-15 feet of panniru verity with a support of forest trees or gliricidia plant, and coffee is an intercropping in sand mixed and black soil.Most of the farmers not aware of pesticide effect on human health.Some of them were engaged in Fig. 3: Aflatoxin sample the application of pesticides for the high yielding purpose and revealed via interview, the generally used pesticides in this district were copper sulphate, bodomixture, trichoderma, copper oxichloride etc. applied through power/hand spraying for controlling black and yellow wilt diseases. 17.This study shows the quantitative estimation of fungicide, pesticide residual content in the collected pepper samples

CONCLUSION
As per the sur vey and quantitative estimation of fungicide(Aflotoxin), pesticide(Organo phosphorous) analysis results shows the samples collected from the different villages of Ponnempet of Kodagu district Karnataka was within the EU limits hence it was free from fungicides, pesticides contents.

Table 3 : Amount of Pesticides present in sample no. 3
Fig.1: Aflatoxin blankAnalysis of Aflatoxin by HPLC