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Under these conditions, the measured TPC was 26.6 mg GAE/L in maceration, 29.8 mg GAE/L in HAE and 32.7 mg GAE/L in UAE, which was in agreement with the predicted values, while TFC was 14.3 mg CE/L, 12.4 mg CE/L and 16.7 mg CE/L for maceration, HAE and UAE, respectively.
For maceration extraction, the material was placed in an Erlenmayer flask, the corresponding amount of solvent was added (details in Table 1) with the sample left to macerate in the dark for 72 h at room temperature.
Stem segments approximately 10 mm in length and 3 mm in diameter were used for maceration.
For maceration, dried powdered leaves (250 g) was extracted with methanol, ethanol, methanol (50%%), ethanol (50%%) and water in triplicate at 25 °C for 72 h.
The dried leaves were further chopped into small pieces and reduced to powder using electronic miller and made suitable for maceration extraction procedure with 80%% methanol.
Small fragments of tissue were treated for maceration procedure (Riva et al, 1999) and studied in a ZEISS Supra40 HR SEM.
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For macerations, small shavings were placed in vials filled with Franklin's solution: glacial acetic acid and 6%% hydrogen peroxide in proportions of 1 : 1 (Franklin 1945).
Crushed shade-dried leaves (500 g) were kept for cold maceration in chloroform: water (1 99) for 4 days at room temperature.
These yields are comparable to the ones recently published for dynamic maceration, but with the advantage of shorter extraction times.
The amount of pigments extracted in the base wine was low as a consequence of the necessity for short maceration time (48 h) and low alcohol content (< 3.5% v/v).
Methods for the maceration of pellets in buffer and plating are detailed in [63].
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