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|Molecular Weight:||121.135||Molecular Formula:||C4H11NO3|
|Appearance:||White Crystal Powder||Storage Conditions:||Room Temperature, Light-proof And Moisture-proof|
77-86-1 Tris Hydroxymethyl Methylamine,
Biological Buffer Tris Hydroxymethyl Methylamine,
77-86-1 Tris Biological Buffer
Tris(hydroxymethyl)aminomethane 77-86-1 Tromethamine use for Biological buffer
How to detect the content of tromethamine (TRIS) in biological products?
Tris(hydroxymethyl)aminomethane (TRIS), also known as tromethamine, is a weak base and is often used as a biological buffer system in the production of biological products. It can cause hypoglycemia, hypotension, nausea, and vomiting. Excessive intake or renal insufficiency can cause alkalemia. It can also inhibit breathing or even stop breathing. Therefore, it is necessary to detect the content of TRIS in biological products. How to detect it?
|PH value required(25℃)||7.10||7.20||7.30||7.40||7.50||7.60||7.70||7.80||7.90||8.00||8.10||8.20||8.30||8.40||8.50||8.60||8.70||8.80||8.90|
|0.1mol/L HCI volume||45.7||44.7||43.4||42||40.3||38.5||36.6||34.5||32||29.2||26.2||22.9||19.9||17.2||14.7||12.4||10.3||8.5||7|
At present, the quantitative detection of TRIS mainly uses acid-base titration, but its limit of quantification is high and it is susceptible to interference from various acid-base groups. Biological products contain a large number of carboxyl groups, amino groups, anions and cations, and it is impossible to quantitatively detect TRIS in products by titration.
To solve the above problems, researchers invented an ion chromatography method to detect TRIS Buffer in biological products. The specific steps are as follows:
(1) Add water for injection to the biological freeze-dried product to dissolve and filter, or directly filter the biological aqueous solution, and take the filtrate as the test solution; add TRIS to the water for injection to dissolve, and obtain solutions of different concentrations as the reference solution after gradient dilution;
(2) Adopt pulsed amperometric detector and post-column fluid replenishment method. The chromatographic column is a CS16 cation exchange column, the mobile phase is 40 mmol/L methanesulfonic acid solution, and the flow rate is 1 mL/min. The post-column replenishment is an alkaline solution of sodium hydroxide or potassium hydroxide with a concentration of 500 mmol/L and a flow rate of 0.3 mL/min;
(3) Quantitative analysis adopts the external standard method to detect the prepared reference solutions of various concentrations, draw the concentration-peak area standard curve of TRIS, and obtain the regression equation of the standard curve; the sample solution is tested under the same conditions, The content of TRIS in the sample is calculated according to the above regression equation. The detection limit was 3.14×10-5mmol/L, the lower limit of quantification was 1.05×10-4mmol/L, the recovery rate was 93.8%~100.1%, and the reproducibility range was 0.10%~0.52%. (RSD, n=5).
Compared with the traditional acid-base titration method, this method has lower detection limits and lower quantification limits, and is not interfered by the acid-base groups carried by the protein, and is not interfered by the anions and cations in the sample, and does not require derivatization and complicated sample preparation. It can detect the content of TRIS (CAS 77-86-1) in different biological products, with good selectivity, and no other peak interference within the TRIS retention time range. With high detection sensitivity and fast analysis speed, it can be applied to the content detection and quality evaluation of TRIS in biological products in related industries.
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