Endotoxin Detection – from Limulus Amebocyte Lysate to Recombinant Factor C

Author(s):  
Jeak Ling Ding ◽  
Bow Ho
2020 ◽  
Vol 16 (2) ◽  
pp. 204-213 ◽  
Author(s):  
Melissa A. Vetten ◽  
Mary Gulumian

Background: Endotoxin-free engineered nanoparticle suspensions are imperative for their successful applications in the field of nanomedicine as well as in the investigations in their toxicity. Gold nanoparticles are known to interfere with various in vitro assays due to their optical properties and potential for surface reactivity. In vitro endotoxin testing assays are known to be susceptible to interference caused by the sample being tested. Objective: This study aimed to identify a preferred assay for the testing of endotoxin contamination in gold nanoparticle suspensions. Methods: The interference by gold nanoparticles on three assays namely, the commonly used limulus amebocyte lysate chromogenic assay, the limulus amebocyte lysate gel-clot method, and the less common recombinant Factor C (rFC) assay, was tested. Results: Possible interference could be observed with all three assays. The interference with the absorbance- based chromogenic assay could not be overcome by dilution; whilst the qualitative nature of the gel-clot assay excluded the possibility of distinguishing between a false positive result due to enhancement of the sensitivity of the assay, and genuine endotoxin contamination. However, interference with the rFC assay was easily overcome through dilution. Conclusion: The rFC assay is recommended as an option for endotoxin contamination detection in gold nanoparticle suspensions.


2008 ◽  
Vol 43 (4) ◽  
pp. 291-303 ◽  
Author(s):  
Ronald Gehr ◽  
Santiago Parent Uribe ◽  
Isabel Fatima Da Silva Baptista ◽  
Bruce Mazer

Abstract Endotoxins are a component of most Gram negative bacteria, and some cyanobacteria. They may be toxic to humans when inhaled or injected, but the effects are unclear when they are ingested. In fact, low concentrations may protect children against certain allergies. Data for endotoxins in Quebec waters are unavailable, hence this study mapped levels in the waters around Montreal, using two commercial test methods. The recently developed factor C method had a greater linear range and was more convenient to use than the widely used Limulus amebocyte lysate (LAL) method. Although the methods gave endotoxin values of the same order, a consistent relationship between the two could not be established. Endotoxin concentrations in the untreated waters varied from 32 to 1,188 EU/mL, comparable in the literature from pristine waters to wastewaters. Values were generally lower in the summer. Filtration is known to be partially effective at removing endotoxins, but the effects of disinfection are not well established. Accordingly, chlorination, ozonation, and ultraviolet light were tested for the destruction of endotoxins in water, at doses found during drinking water disinfection. While chlorine and ultraviolet light had minimal effects on endotoxin levels, ozone could achieve up to 60% reductions at Ct values (concentration x contact time) as low as 2.5 mg•min/L.


2020 ◽  
Vol 3 (2) ◽  
pp. 115
Author(s):  
Andrea Santosa ◽  
Ida Bagus Made Artadana ◽  
Mariana Wahjudi

Peningkatan kualitas produk industri farmasi mendorong tersedianya uji yang akurat dan efisien untuk mendeteksi adanya pirogen. Endotoksin merupakan salah satu dari pirogen yang umumnya ditemukan sebagai kontaminan pada produk obat. Hingga saat ini pengujian pirogen yang digunakan secara umum di Indonesia adalah metode Rabbit Pyrogen Test (RPT) dan Limulus Amebocyte Lysate (LAL). Namun, kedua metode ini memiliki kekurangan yang hampir sama, yaitu penggunaan hewan hidup sebagai bahan uji. Untuk alasan ini, saat ini sedang dikembangkan metode in vitro yang lebih menguntungkan untuk uji pirogen, yaitu dengan Recombinant Factor C (rFC) dan Monocyte Activation Test (MAT). Tujuan dari review ini adalah memberikan pemaparan mengenai kelebihan dan kekurangan metode uji pirogen dan endotoksin yang sesuai dengan kebutuhan konsumen, serta kajian penggunaan metode alternatif pada industri farmasi di Indonesia. Diharapkan dengan pemaparan dan kajian yang diberikan dapat digunakan sebagai pertimbangan penggunaan metode uji pirogen dan endotoksin yang lebih efektif dan efisien.


Author(s):  
Maik Luu ◽  
Rossana Romero ◽  
Jasmin Bazant ◽  
Elfadil Abass ◽  
Sabrina Hartmann ◽  
...  

2012 ◽  
Vol 50 (01) ◽  
Author(s):  
C Trierweiler ◽  
K Willim ◽  
HE Blum ◽  
P Hasselblatt

2012 ◽  
Vol 224 (03) ◽  
Author(s):  
I Kuznetsova ◽  
K Welte ◽  
J Skokowa

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