Objective: To establish a quantitative detection method of bacterial endotoxin in umbilical cord mesenchymal stem cell suspension and verify its feasibility. Methods: According to the standards for photometric determination of bacterial endotoxin in General notice 1143 of Chinese Pharmacopoeia part Ⅳ (2020 edition). A preliminary detection method of bacterial endotoxin content in umbilical cord mesenchymal stem cell suspension was established from two aspects: standard curve establishment, reliability verification, sample interference screening and interference detection. Bacterial endotoxins in three batches of umbilical cord mesenchymal stem cell suspensions were quantitatively detected by this method. Results: In the establishment of standard curve and reliability verification test, the linear regression equation was lgT=2.778 0-0.281 4lgC, r=-0.999 8,|r|>0.980, negative control time>3 600 s (standard curve low average response of time was 2 690 s), repeated reaction tube RSD were 10% or less, the standard curve was established successfully. The results of interference screening test showed that the stem cell suspension had no interference on limulus test at the dilution ratio of 20, 40 and 80, and the recovery rate was close to 100% at the dilution ratio of 40, indicating that 40 times was the best interference dilution ratio without interference. The results of the three batches of samples showed that the endotoxin content of the three batches of samples were less than 0.5 EU·mL-1 under 40 times dilution, and the RSD was less than 10%. The positive recovery was 86.8%-101.2%, which complied with The Chinese Pharmacopoeia 2020 edition. Conclusion: This method can quickly and quantitatively detect the content of endotoxin in umbilical cord mesenchymal stem cell suspension, and has strong anti-interference ability, which can meet the determination of bacterial endotoxin in cell suspension.
YUAN Chao-lu, ZHANG Si-shi, CHEN Yi-ru, LI Jia-hui, TANG Jia
. Dynamic chromogenic method for detection of bacterial endotoxin content in umbilical cord mesenchymal stem cell suspension[J]. Chinese Journal of Pharmaceutical Analysis, 2022
, 42(12)
: 2110
-2115
.
DOI: 10.16155/j.0254-1793.2022.12.06
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