Derivatization (유도체화)

* 시료 유도체화(Derivatization)

 - Reference

  • https://www.sciencedirect.com/science/article/pii/S0168165621001474#bib0080

  • Before GC/MS analysis, we derivatized the extracted metabolites by methoxyamination and trimethylsilylation (Lee and Fiehn, 2008).

  • The methoxyamination step began as we added 10 μL of 40 mg/mL methoxyamine chloride in pyridine (Sigma-Aldrich, St. Louis, MO, US)

     to the samples and then incubated the samples for 90 min at 30 °C, shaking at 200 rpm.

  • Then, we added 45 μL of N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA; Sigma-Aldrich) to the samples for trimethylsilylation and

     incubated for 30 min at 37 °C and 200 rpm shaking.

 

 - 재료 : sample, Methoxyamine hydrochloride, Pyridine, N-methyl-N-trimethylsilyl trifluoroacetamide (MSTFA)

 

 - Process : methoxyamination, trimethylsilylation으로 유도체화

  <Methoxyamination step>

  • ① Methoxyamine hydrochloride 40mg(0.04g)에 Pyridine 1ml를 첨가한다. (한번에 미리 만들어 두면 석출되므로 필요할 때마다 제조한다.)

  • ② ①의 용액 중 10 𝜇L를 sample에 첨가한다.

  • ③ Vortexing, Centrifuge를 진행한 후 30℃ 에서 90분간 1000 rpm으로 shaking한다.

  <Trimethylsilylation step>

  • ④ ③의 용액에 45uL의 N-methyl-N-trimethylsilyl trifluoroacetamide을 추가하고 37℃에서 30분간 1000 rpm으로 shaking한다.

 

* Names of conventional derivatizing agents

 - Reference

  • https://link.springer.com/referenceworkentry/10.1007/978-94-007-7864-1_71-1

 

Method	Reagent (시약)	Common name	통칭
Silylation (실릴화)	BSA	Bistrimethylsilylacetamide	비스트리메틸실릴아세트아마이드
	BSTFA	Bistrimethylsilyltrifluoroacetamide	비스트리메틸실릴트리플루오로아세트아마이드
	HMD	Hexamethyldisilane	헥사메틸디실란
	MSTFA	N-methyl-trimethylsilyltrifluoroacetamide	N-메틸-트리메틸실릴트리플루오로아세트아미드
	MTBSTFA	N-methyl-N-t-butyldimethylsilyltrifluoroacetamide	N-메틸-Nt-부틸디메틸실릴트리플루오로아세트아미드
	TMCS	Trimethylchlorosilane	트리메틸클로로실란
	TMS-DEA	Trimethylsilyldiethylamine	트리메틸실릴디에틸아민
	TMSI	Trimethylsilylimidazole	트리메틸실릴이미다졸
Acylation (아실화)	HFBA	Heptafluorobutyric Anhydride	헵타플루오로부티르산 무수물
	HFBI	Heptafluorobutyrylimidazole	헵타플루오로부티릴이미다졸
	MBTFA	N-Methyl-bis(trifluoroacetamide)	N -메틸-비스(트리플루오로아세트아미드)
	PFBCI	Pentafluorobenzoyl Chloride	펜타플루오로벤조일 클로라이드
	PFPA	Pentafluoropropionic Anhydride	펜타플루오로프로피온산 무수물
	PFPI	Pentafluoropropionylimidazole	펜타플루오로프로피오닐이미다졸
	TFAA	Trifluoroacetic Anhydride	트리플루오로아세트산 무수물
	TFAI	Trifluoroacetylimidazole	트리플루오로아세틸이미다졸
Alkylation (알킬화)	BF3(BCl3)	in methanol or butanol	-
	DMA	dimethylacetal	디메틸아세탈
	PFBBr	Pentafluorobenzyl bromide	펜타플루오로벤질 브로마이드
	TBH	Tetrabutylammonium hydroxide	수산화테트라부틸암모늄