- The document discusses nicotine testing methods and their use in various contexts like insurance, employment, schools, and research. It also covers nicotine metabolism and the differences between nicotine and its metabolite cotinine. - The Nano-Check Rapid Nicotine Test is described as an immunoassay that detects cotinine in urine at cutoff levels of 200ng/ml or 500ng/ml to determine tobacco use. It provides results in 5-10 minutes and is intended for professional use. - The test works by detecting cotinine in a urine sample via monoclonal antibodies on a test strip, with positive or negative results depending on whether or not a test line appears within the timeframe.

1. NANO-CHECK TM RAPID NICOTINE TEST NANO-DITECH CORPORATION

5. Latest Tobacco Trends (continued)

15. Procedure of the Test 1 . Remove the test device from the sealed pouch immediately before use 2. Put 80 ul of urine sample using enclosed dropper or pippette into the sample well 3. Wait 5 to 10 min for the result

16. Interpretation of the Results Negative : Two colored lines will appear. The line in the test region (T) is the drug probe line; the line in the control region (C) is the control line, which is used to indicate proper performance of the device. The color intensity of the test line may be weaker or stronger than that of the control line. Negative result does not indicate that the absolute absence of drugs in specimen, it only indicates the specimen does not contain the drug at a concentration equal or above the cut off level in qualitative terms. Positive : One colored line appears on the control region ( C ), the absence of test line ( T ) is a presumptive positive result for cotinine; i.e. the specimen may contain cotinine or its analogous compounds at concentration above the cutoff level. Invalid : If no colored line in the control region, the test result is invalid. Under no circumstance should a positive sample be identified until the control line appears in the viewing area. If the colored line does not appear, the test result is inconclusive and the assay should be repeated with a new test device. Negative Positive Invalid

17. Sensitivity The Nano-Check TM Rapid Nicotine test detects cotinine in human urine at the cut-off concentrations of 200ng/ml or 500ng/ml. The cutoff value was determined using control urines spiked with the specified compounds at the indicated concentration; 25%, 50 % below cut off level, and 25%, 50% above cut off level. Cotinine Level (ng/ml) Negative. Positive Coincidence (%) 100 25 0 100 150 25 0 100 200 15 10 N/A 250 4 21 84 500 15 10 N/A

18. Accuracy The Nano-Check TM Rapid Nicotine test detects cotinine in human urine at the cut-off concentrations of 200ng/ml or 500ng/ml. The cutoff value was determined using control urines spiked with the specified compounds at the indicated concentration; 25%, 50 % below cut off level, and 25%, 50% above cut off level. GC/MS Results Positive Negative Total Nano-Check TM Rapid Nicotine Result Positive 49 0 49 Negative 1 100 101 Total 50 100 150 Agreement 98.0 % 100 % 99.3 %

19. Specificity & Cross Reactivity The following compounds were tested to determine cross reactivity at the cut off concentration of target drug with Nano-Check TM Rapid Nicotine test . Compounds Cutt-off (ng/ml) Cross Reactivity Nano-Check TM Rapid Nicotine Result Cotinine 200 100 Nicotine > 100,000 < 0.2 Niacinamide > 100,000 < 0.2 Nicotinic Acid > 100,000 < 0.2

20. Thank you for your attention Contact Pammvi Group of Companies for more information about pricing and availability C 501 Remi Bizcourt Off Veera Desai Andheri West Mumbai 400053 India Telephone nos 91 22 26731904 / 26731052 / 26732760 Fax 91 22 26731901 Website www.pammvi.com email id [email_address]