Full
Outline
To determine of γ-aminobutyric acid (GABA) in suancai fermentation broth by high performance liquid chromatography with diode array detection (HPLC-DAD) coupled with pre-column derivatization with dansyl chloride.
In this study, HPLC-DAD was used to perform real-time online spectral scanning of derivatives. Combined with single factor experimental method, the derivatization reaction conditions and chromatographic separation parameters were systematically optimized.
Using NaHCO3-Na2CO3 as buffer system, 45 ℃ water bath derivatization reaction for 5 min, and without ammonia, the derivatization effect was the best, but the sample pH still needed to be adjusted to more than 10 when determining the suancai fermentation broth. The mobile phase with 0–15 min, 30%–42% acetonitrile, 70%–58% 20 mmol/L phosphoric acid could not only achieved a stable baseline, but also made the target and interferents baseline separation. when the organic mobile phase changed from methanol to acetonitrile, the derivative λmax changed from 219 nm to 222 nm, that was the λmax shifted red. At the wavelength of λmax(222 nm), coefficient of determination reached 0.99999, with the highest sensitivity and the lowest limit of detection (0.023 mg/L), which revealed that λmax was the optimal detection wavelength. The method validation results showed that the optimized method exhibited a good linear relationship within the mass concentration range of 0.05–100.00 mg/L. The limit of detection of the method was 0.074 μg/mL, and the limit of quantitation was 0.241 μg/mL. The recovery rate was 87.02%–107.25%, with a relative standard deviation of 2.34%–3.49%. Furthermore, it was found that GABA had poor stability and should not be stored for extended periods even under refrigeration.
The optimized method can be used to determine GABA in suancai fermentation broth with complex matrices, demonstrating good sensitivity, precision and reproducibility. It is also confirmed that ammonia can induce derivative side reactions, changes in the organic mobile phase can cause changes in λmax, and the GABA has poor stability.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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