Characteristics

• • Pyrosep^® adsorbs and concentrates endotoxins in the sample, so it is possible to specifically measure very small amounts of endotoxins without being affected by endogenous materials.
• • The test can be applied to fatty materials such as fat-soluble vitamins if the material is soluble in ethanol because Pyrosep^® works as affinity resin to endotoxin even in ethanol.
• • The test is no waste because LAL reagent is efficiently used, depending on the numbers of samples to be assayed.

Measurement principle

Endotoxin in the sample is specifically concentrated by passing the sample through Pyrosep^® (An affinity adsorbent in which histidine is bound to a water-insoluble carrier via a spacer, specifically adsorbing endotoxin) packed in glass capillary column. Absorbed endotoxin on the resin is directly conducted to LAL ES reagent and the coagulation level specifically induced by endotoxin is measured with a Toxinometer^®.

Content

1. 1. LAL ES reagent, Limulus amebocyte lysate lyophilized 20 vials （for 0.3 mL）
2. 2. Pyrosep^® Suspension （containing in phosphate buffer） 20 vials （0.77 mL）
3. 3. LAL reconstitution solution 2 vials （ 7 mL）
4. 4. Washing solution 4 vials （11 mL）
5. 5. Sample Dilution Buffer Solution 4 vials （10 mL）
6. 6. Capillary Endotoxin Free （having a filter at the connected end with silicon tube, endotoxin free） 20 pieces

※This kit does not contain syringe, 3 way stopcock and syringe fitting

Method

Application example

A. Application to the measurement of endotoxin in dialysis fluid

After adding USP standard endotoxin to 5.0 mL of each of the commercially available peritoneal dialysis fluid Dianeal PD4 (Baxter Japan K.K.) and SUBLOOD^® A and B Substitution Fluid for Hemofiltration BSG (Fuso Pharmaceutical Industries, Ltd. ), the pH was adjusted to around 5 using 0.1 M acetic acid to prepare the samples. The endotoxin concentration of these samples was measured using the Pyrosep^® method, and the recovery rate of the added endotoxin was determined.
Using the Pyrosep^® method, endotoxin in the dialysis fluid could be detected at a concentration of 0.1 EU/L.

B. Application of endotoxin measurement in fat-soluble vitamins

After dissolving the fat-soluble vitamins, vitamin D, E and K, in ethanol and adding USP standard endotoxin,the endotoxin concentration was measured using the Pyrosep^® method, and the recovery rate of the added endotoxin was determined.
By using the Pyrosep^® method, it is now possible to measure endotoxin in fat-soluble vitamins, which was previously impossible using Limulus reagents.

C. Other application examples

Ethanol, silicone oil, soybean oil, olive oil, stearic acid, egg yolk lecithin, oil adjuvant vaccines, albumin formulations, coagulation factor formulations (Factor VIII), interferon formulations, antithrombin III formulations, immunoglobulin formulations, and other samples that can be dissolved in ethanol can be measured.