Ultrahigh performance liquid chromatography-tandem mass spectrometry determination of glutamine in human assisted reproductive medium
10.3969/j.issn.2095-4344.1617
- VernacularTitle:超高效液相色谱-串联质谱法测定辅助生殖用培养液中谷氨酰胺的含量
- Author:
Linnan KE
1
;
Yuanli HUANG
;
Yu FANG
;
Chunren WANG
Author Information
1. 中国食品药品检定研究院
- From:
Chinese Journal of Tissue Engineering Research
2019;23(10):1594-1598
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Ultrahigh performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for analyzing amino acids in biological samples is simple in sample preparation with a short analysis time, and has high sensitivity and specificity. Until now, it is only applied in analyzing glutamine contents in dialysate, urine and plasma. OBJECTIVE: To establish a method for determining glutamine concentration in human assisted reproductive media by UPLC-MS/MS. METHODS: The UPLC separation was performed on a SUPELCO Discovery HS F5-3 column (15 cm×2.1 mm, 3 μm) at 40 oC. The mobile phase was acetonitrile and water (both containing 0.1％ formic acid) in a gradient elute mode. The flow rate was 0.35 mL/min. Electrospray ionization with a negative-ion and multiple reaction monitoring mode was used. RESULTS AND CONCLUSION: The linearity was achieved in the range of 0.123 7-24.74 mg/L for glutamine (r=0.999 7). The recoveries were 102.9％-108.2％ with the range 2.3％-4.9％ for the relative standard deviation. The limit of qualification was 9.76 μ g/L. The fertilization culture medium containing glutamine was incubated at 37 oC for 96 hours, in which the glutamine content declined 6％ at 24 hours and 15％ at 96 hours respectively compared with initial content. Therefore, the method is simple, specific, accurate and sensitive without sample derivation, and the test time is short. It is suitable for the quality control of human assisted reproductive media and useful for the risk study related to the degradation of glutamine.
