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辅酶A系列产品色谱方法说明
发布时间:2020-08-18 13:51 | 点击次数:
关于ZL-23010 – ZL-23078 的辅酶A系列产品,国外建议如下:
辅酶A衍生物不建议使用普通的RP C18色谱柱进行分析,因这类化合物会粘附在色谱柱上,难以从色谱柱中洗脱出来。
色谱条件建议如下:
Chromolith Type色谱柱
洗脱剂:10%溶剂B(yi腈/jia醇)+ 90%溶剂A(0.1%jia酸水溶液)
若您选取的是普通的C18色谱柱,溶剂A务必调整为0.2% 三氟yi酸(TFA) 水溶液即每1L水中三氟yi酸用量2mL (溶剂A)。
梯度洗脱 溶剂A:0.2%TFA水溶液 溶剂B:yi腈/jia醇
0 - 8 min A:B = 80:20
8 - 9 min B = 100%
9 - 10 min A:B = 80:20
国外关于该类产品的原文说明如下:
Please tell the customer he should read the Certificate of analysis with attention,
because the Coenzyme-A-Derivatives can NOT be tested with usual, ordinary RP C18 columns, because the compound is stick (adhere) on the column an it is NOT eluting from the column !!!
For getting elution, the customer need to use Chromolith Type columns with 0.1% Formic Acid in Water and starting HPLC-MS conditions with 10% Solvent B (Acetonitril or MeOH) and 90% Solvent A (Water with 0.1% Formic Acid).
In case of using common RP C18 columns, proceed as following: Use as Solvent A 0.2% TFA (Trifluoroacetic Acid - this is meaning 2ml TFA per Liter Water) and starting HPLC-MS conditions using 20% Solvent B (Acetonitril or MeOH) and 90% Solvent A (Water with 0.2% Trifluoroacetic Acid) than increase in 8 minutes the Solvent B to 100%, keep 1 minute at 100% and go back to 20% Solvent B in 1 Minute keeping this gradient for another minute.