A method for simultaneous detection of carboxymethyllysine and carboxyethyllysine in green tea
A technology of carboxymethyllysine and carboxyethyllysine, which is applied in the field of analysis and detection, can solve problems affecting the applicability of the method, and achieve good stability
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Embodiment 1
[0034] (1) Extraction: Weigh 50 mg tea powder sample into a 10 mL centrifuge tube, add 2 mL of n-hexane, shake vigorously for 3 min, then centrifuge at 10,000 rpm for 5 min, discard the solvent, and repeat the above steps twice. After degreasing with n-hexane, the sample was blown dry with nitrogen until it recovered to a powder state. Add 1mL Na to the sample 2 B 4 o 7 Buffer (0.2mol / L) and 0.5mL NaBH 4 (1mol / L, prepared with 0.1mol / L NaOH), adjust pH=9.2, and reduce at 4°C overnight (about 8h). Next, add 1 mL of chloroform / methanol (2:1, v / v), centrifuge at 10,000 rpm for 10 min, discard the supernatant; add 5 mL of HCI (6 mol / L) to the obtained precipitate, and place it in an oven at 110 ° C , Hydrolyzed for 12h. After the hydrolysis was completed, the hydrolyzed sample was taken out and cooled to room temperature (25°C), and the volume was adjusted to 10 mL with ultrapure water, then filtered with qualitative filter paper and the filtrate was collected.
[0035] (2) ...
Embodiment 2
[0044] (1) Extraction: Weigh 50 mg of tea powder (same as in Example 1) and place it in a 10 mL centrifuge tube, add 2 mL of n-hexane, vibrate vigorously for 3 min, then centrifuge at 5000 rpm for 10 min, discard the solvent, and repeat the above steps 3 times. After degreasing with n-hexane, the sample was blown dry with nitrogen until it recovered to a powder state. Add 1mL Na to the sample 2 B 4 o 7 Buffer (0.1mol / L) and 0.5mL NaBH 4 (0.8mol / L, prepared with 0.1mol / L NaOH), adjust pH=8.5, and reduce at 4°C for 10h. Next, add 1 mL of chloroform / methanol (1:1, v / v), centrifuge at 5000 rpm for 10 min, discard the supernatant; add 5 mL of HCI (4 mol / L) to the obtained precipitate, and place in an oven at 100 ° C , Hydrolyzed for 30h. After the hydrolysis was completed, the hydrolyzed sample was taken out and cooled to room temperature (25°C), and the volume was adjusted to 10 mL with ultrapure water, then filtered with qualitative filter paper and the filtrate was collecte...
Embodiment 3
[0054] (1) Extraction: Weigh 50 mg of tea powder (same as in Example 1) and place it in a 10 mL centrifuge tube, add 2 mL of n-hexane, vibrate vigorously for 3 min, then centrifuge at 12000 rpm for 4 min, discard the solvent, and repeat the above steps twice. After degreasing with n-hexane, the sample was blown dry with nitrogen until it recovered to a powder state. Add 1mL Na to the sample 2 B 4 o 7 Buffer (0.3mol / L) and 0.5mL NaBH 4 (1.2mol / L, prepared with 0.1mol / L NaOH), adjust pH=10, and reduce at 6°C for 2h. Next, add 1 mL of chloroform / methanol (3:1, v / v), centrifuge at 12000 rpm for 4 min, discard the supernatant; add 5 mL of HCI (8 mol / L) to the obtained precipitate, and place in an oven at 120 ° C , Hydrolyzed for 12h. After the hydrolysis was completed, the hydrolyzed sample was taken out and cooled to room temperature (25°C), and the volume was adjusted to 10 mL with ultrapure water, then filtered with qualitative filter paper and the filtrate was collected. ...
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