Chromium Assay Kit, BioAssay(TM)

Chromium is widely used in various industries such as electroplating, leather tanning, chrome paint, dying, hardened steel, ceramic and glass industry. Chromium exists in two stable oxidation states, hexavalent Cr(VI) and trivalent Cr(III). Cr(VI) is produced solely by industrial processes, whereas in nature, chromium exists in its trivalent form. Cr(III) is generally regarded as nontoxic due to poor absorption. Cr(VI) is considered a pulmonary carcinogen and has tested positive in genotoxicity tests. It is one of the most serious pollutants in many water streams due to its carcinogenic potential. Most countries apply a legal limit of 50-100 ug/L Cr in drinking water.\nChromium Assay Kit provides a simple one-step colorimetric means to directly measure Cr(VI) in a sample. In the assay, Cr(VI) forms a stable complex with a specific chromogenic dye. The optical density at 480nm is directly proportionate to the Cr(VI) concentration in the sample. Cr(III) can be converted to Cr(VI) with nitric acid/hydrochloric acid, thus allowing the determination of Cr(III) or total Cr [Cr(III)+Cr(VI)] in the sample. The assay is sensitive with a detection limit of 20 ug/L Cr.\n\nKey Features:\nSensitive and accurate. Linear detection range of 20-2000 ug/L\nChromium.\nConvenient and high-throughput. Homogeneous "mix-incubate-measure" type assay. No wash and reagent transfer steps are involved. Can be readily automated for processing thousands of samples per day.\n\nApplications:\nDetermination of chromium in biological (serum, plasma etc), environmental (water, soil etc), food and beverage samples.\n\nKit Contents: (sufficient for 250 tests in 96-well format)\nReagent A: 300ul Reagent B: 20ml\nCr(VI) Standard: 300ul 40 mg/L\nStorage conditions: store Reagent A at -20 degrees C and other reagents at 2-8 degrees C. Shelf life of 6 months after receipt.\nPrecautions: reagents are for research use only. Normal precautions for laboratory reagents should be exercised while using the reagents. Please refer to Material Safety Data Sheet for detailed information.\n\n96-Well Assay Procedure:\nUse clear flat-bottom plates. Prior to assay, bring all reagents to room temperature. Unused Reagent A should be stored at -20 degrees C. Samples should be clear, colorless and free from particles or precipitates. Substances that may potentially interfere with the assay include: azide, Ba2+, Pb2+, Fe3+, Gold(III), Sn(II), Ti (IV). If necessary, water samples can be concentrated by evaporation. If determination of Cr(III) or total Cr [Cr(III)+Cr(VI)] is desired, please refer to the General Sample Treatment Procedure.\n1. Standards. Prepare 600ul 2000 ug/L Cr(VI) Standard Premix by mixing 30ul Standard and 570ul deionized water dH2O (>18 megaohm).\nDilute standard as follows.\nNo Premix+dH2O Standard (ug/L)\n1 300ul+0ul 2000\n2 150ul+150ul 1000\n3 75ul+225ul 500\n4 0ul+300ul 0\nTransfer 250ul standards into separate wells of the plate.\nSamples: transfer 250ul of each sample into separate wells of the plate.\n2. Assay. Prepare enough Working Reagent, for each well, by mixing 1ul Reagent A and 55ul Reagent B. Add 50ul Working Reagent to each well. Tap plate to mix. Incubate for 20 min at room temperature. Read optical density at 480 nm (430-505nm).\n\nCuvette Assay Procedure:\nThe cuvette Assay Procedure: is essentially the same as the 96-well plate assay. The Working Reagent is prepared by mixing 4ul Reagent A and 220ul Reagent B. Assay by mixing 1000ul Standard or Sample with 200ul Working Reagent.\n\nCalculation:\nSubtract the blank control OD (#4) from the OD values of the standards. Plot the Standard Curve and determine its Slope. Cr(VI) concentration of a Sample is calculated as,\n[Cr(VI)] =\nODSAMPLE

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SPECIFICATIONS

Catalog Number

C5069-95W

Size

1Kit

References

1. Barceloux, DG (1999). Chromium. J Toxicol Clin Toxicol. 37(2): 173-94.\n2. Greenberg, AE, Clesceri, LS, Trussell, RR, Eds. (1995) Standard Methods for the Examination of Water and Wastewater, American Public Health Association, 19th ed., Washington, DC, 3-59.\n3. De, AK (2000). Environmental Chemistry, 4th ed.; New Age International, New Delhi, 229, 2000.