Citation: LONG Zhen, WEI Chen, GUO Zhimou, MA Xiao, LI Yueqi, YAO Jinting, JI Feng, LI Changkun, HUANG Taohong. Determination of tracheal cytotoxin in pertussis and diphtheria tetanus acellular pertussis vaccines using liquid chromatography-tandem mass spectrometry[J]. Chinese Journal of Chromatography, ;2019, 37(2): 155-161. doi: 10.3724/SP.J.1123.2018.08006 shu

Determination of tracheal cytotoxin in pertussis and diphtheria tetanus acellular pertussis vaccines using liquid chromatography-tandem mass spectrometry

LONG Zhen ^1, ,
WEI Chen ² ,
GUO Zhimou ³ ,
MA Xiao ² ,
LI Yueqi ¹ ,
YAO Jinting ¹ ,
JI Feng ¹ ,
LI Changkun ¹ ,
HUANG Taohong ¹

1.
Shimadzu Scientific Instrument Company, Beijing 100020, China;
2.
National Institutes for Food and Drug Control, Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products, Beijing 102629, China;
3.
Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China

Received Date: 22 August 2018

Tracheal cytotoxin (TCT) is a toxic glycopeptide, which contribute to the adverse effects of pertussis toxin (PT) and related vaccines. Although pharmacopeias limit the amount of TCT in PT product, there is no recommended TCT determination method in any pharmacopeia. In this study, a liquid chromatography-tandem mass spectrometry method was developed to determine TCT. Chromatographic conditions, including column-type and mobile-phase composition, were optimized. According to the literature reports, both reversed-phase liquid chromatography (RPLC) and hydrophilic interaction liquid chromatography (HILIC) can provide a good retention for TCT. A large amount of organic solvent is usually used for protein precipitation, which may affect the RPLC mode, leading to peak distortion, while such effects were not observed in HILIC mode. Thus, HILIC mode was used to analyze TCT in this study. The developed method had a wide linear range (5.76-369 ng/L), good precision (no more than 3.9%), satisfied recoveries in various matrices (96.4%-102.5%). The limit of quantification (LOQ) of the developed method was 1279 times lower than the one required by Chinese Pharmacopeia, wherein the required amount of TCT should be less than 2 pmol per dose. The developed method was used to detect TCT in pertussis vaccine (acellular component), pertussis vaccine (acellular, co-purified), co-purified diphtheria tetanus pertussis vaccine, and component diphtheria tetanus acellular pertussis vaccine. As a result, TCT was not detected in any of the selected samples indicating the safety of these vaccines and PT products.
- liquid chromatography (LC),
- mass spectrometry (MS),
- tracheal cytotoxin (TCT),
- diphtheria tetanus pertussis acellular vaccine (DTaP),
- pertussis vaccine (PT)
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