Objective: To establish the first national standard for oncolytic activity assay of herpes simplex virus type 1(HSV-1). Methods: According to the requirements in Chinese Pharmacopoeia(Volume Ⅲ, 2020 edition), the liquid and freeze-dried standard for oncolytic activity of HSV-1 were prepared and tested,of which the stability were evaluated by thermal acceleration test. The oncolytic activity of the standard was calibrated collaboratively by U-2 OS cells/CCK-8 method in 3 laboratories. The liquid standard was compared with the freeze-dried standard, and the more suitable one was selected as the national standard. Results: The prepared standard substance was all qualified, among which the moisture content of the freeze-dried standard was 1.09% and the dispensing accuracy was 0.15%. The results of stability test were calculated by Arrhenius formula. It was preliminarily predicted that it would take 7.7 years for the oncolytic activity of liquid standard to decrease by 10% at -70 ℃, and it would take 6.1×105 years for the oncolytic activity of lyophilized standard to decrease by 10% at -70 ℃. Compared with liquid standard, the stability of lyophilized standard was greatly improved. Twenty-one times of collaborative calibration tests by 3 laboratories showed that the oncolytic activity liquid standard was 7.08×104 U·mL-1 and the oncolytic activity liquid standard was 1.82×104 U·vial-1. After lyophilized, the oncolytic activity of the bulk of HSV-1 standard decreased from 7.08×104 U·mL-1 to 3.03×104 U·mL-1. However, the good S-shaped dose-response curve still appeared after 100 times of pre-dilution, which did not affect the requirements of its use as a standard. When the liquid standard was used as the sample and the freeze-dried standard was used as the standard for calibration, the geometric coefficient of variation (GCV) of the results of the 3 laboratories decreased from 64.4% to 29.2%, and the precision of the experiment was greatly improved. Conclusion: The batch of freeze-dried HSV-1 standards for oncolytic activity assay meets the relavant requirements, and is more suitable for use as a national standard than liquid standards. Its oncolytic activity is assigned a value of 1.82×104 U·vial-1.
QIN Xi, HU Jin-pan, LI Yong-hong, SHI Xin-chang, DING You-xue, BI Hua, HAN Chun-mei, ZHENG Hong-mei, RAO Chun-ming, LIANG Cheng-gang
. Preparation of the candidate first Chinese national standard for oncolytic activity assay of herpes simplex virus type 1*[J]. Chinese Journal of Pharmaceutical Analysis, 2024
, 44(4)
: 663
-670
.
DOI: 10.16155/j.0254-1793.2024.04.13
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