Description
Salicylate Assay Kit (BA0225) (BA0225)
The Salicylate Assay Kit (SKU: BA0225) provides a convenient and reliable colorimetric method for the quantitative determination of salicylate. Salicylate is a salt or ester of salicylic acid found naturally in some plants and is also a metabolic byproduct of aspirin, so its concentration is often measured in blood or urine in cases of suspected overdose. In the assay, salicylate complexes with ferric chloride to create a coloured compound measured at 560 nm. The assay is simple, sensitive and adaptable to high-throughput screening, with a linear detection range of 0.8 mM (10.9 mg/dL) to 20 mM (274.2 mg/dL) salicylate using 20 microlitres of sample in a 96-well plate. It can be used with a wide variety of samples including serum, plasma, urine, beauty products and mouthwash, and the add-mix-read format can be readily automated for thousands of samples per day.
| Product Name: | Salicylate Assay Kit (BA0225) |
| SKU: | BA0225 |
| Detection Method: | Colorimetric; read at 560 nm (500-600 nm) |
| Detection Range: | 0.8 mM (10.9 mg/dL) to 20 mM (274.2 mg/dL) salicylate (96-well) |
| Sample Type: | Serum, plasma, urine, beauty products, mouthwash |
| Species Reactivity: | All |
| Assay Time: | Approximately 20 minutes |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | Store kit at 2 degrees C to room temperature. |
| Shelf Life: | 12 months after receipt |
| Shipping: | Room Temperature |
Salicylate is a salt or ester of salicylic acid and can be found naturally in some plants. It is also a metabolic byproduct of aspirin (acetylsalicylic acid), and salicylate concentrations are often tested in blood or urine in cases of suspected overdose. Salicylic acid is commonly used in skincare products as an exfoliating ingredient and in other consumer products as a preservative. In this assay, salicylate complexes with ferric chloride to create a coloured compound measured at 560 nm; the assay can be used with a variety of samples and is simple, sensitive and adaptable to high-throughput screening.
- Fast and sensitive: linear detection range of 0.8 mM (10.9 mg/dL) to 20 mM (274.2 mg/dL) salicylate with 20 microlitres of sample in a 96-well format.
- Convenient: the procedure involves adding a single working reagent.
- High-throughput: add-mix-read type assay that can be readily automated as a high-throughput 96-well or 384-well plate assay for thousands of samples per day.
- Direct assays of salicylate in biological samples (e.g. serum, plasma, urine).
- Determination of salicylate in consumer products such as beauty products and mouthwash.
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
| Step | Procedure |
| 1 | For sample preparation, ensure samples are transparent and precipitate-free; if cloudy or containing precipitates, centrifuge 5 minutes at 14,000 x g and use the clear supernatant. Serum, plasma, urine and other liquid samples can be used directly, but high-protein samples may precipitate due to the acidity of the Reagent, in which case combine the sample and Reagent in a microcentrifuge tube, mix well, centrifuge and use the supernatant. |
| 2 | Prepare 200 microlitres of 20 mM Premix by mixing 40 microlitres of the 100 mM Standard with 160 microlitres of distilled water, then dilute standards in 1.5 mL centrifuge tubes as described in the standard preparation table. |
| 3 | Transfer 20 microlitres of standards into separate wells of a clear flat-bottom 96-well plate, transfer 20 microlitres of a sample into a single well and 20 microlitres into another well for the sample blank. |
| 4 | Add 180 microlitres of Reagent to each Standard and Sample well, add 180 microlitres of deionised water to the Sample Blank well and tap the plate lightly to ensure the contents are mixed evenly. |
| 5 | Read optical density at 560 nm (500-600 nm). |
| 6 | Cuvette procedure: dilute standards three-fold by adding 40 microlitres of standard to 80 microlitres of water in tubes, transfer 100 microlitres of standards and samples to cuvettes, add 300 microlitres of Reagent and 600 microlitres of water to a final volume of 1000 microlitres in each cuvette, then read optical density at 560 nm (500-600 nm). Note: with a 1 mL cuvette the linear detection range changes to 0.4 mM - 10 mM because cuvettes have higher pathlengths. |
Subtract the Blank value (Standard #4) from the standard values and plot the delta-OD against standard concentrations. Determine the slope and calculate the salicylate concentration as: [Salicylate] = [(ODSAMPLE - ODSAMPLE BLANK) / (ODSTANDARD - ODSAMPLE)] x ([Standard]/4) x n (mM), where ODSAMPLE and ODSAMPLE BLANK are the OD560 readings of the Sample and Sample Blank respectively and n is the dilution factor. Note: if the calculated salicylate concentration is higher than 20 mM, dilute the sample in water and repeat, multiplying the result by the dilution factor n. Conversions: 1 mM Salicylate is 137.11 ppm, 13.7 mg/dL, 0.014% w/v.
| Component | Quantity | Storage |
| Reagent | 20 mL | 2 degrees C to RT |
| Standard (100 mM salicylate) | 200 uL | 2 degrees C to RT |
