Abstract: The content of ferulic acid, protocatechuic aldehyde and anthocyanin in sorghum grains was determined by high-performance liquid chromatography (HPLC). The content of total phenol, total flavonoids and condensed tannin was determined by colorimetry; Multiplication establishes a multivariate calibration model between NIR spectroscopy and HPLC and colorimetric analysis values ​​to predict the content of the main phenols in sorghum grains.
1 Introduction
Phenolic compounds are an important secondary metabolic substance in sorghum seeds, mainly phenolic acids, flavonoids and condensed tannins (proanthocyanidins) [1]. Sorghum phenols are not only an excellent natural antioxidant [2], but also have a variety of health care functions and pharmacological effects [3,4]. The conventional quantitative detection methods of phenols include chemical determination and high pressure liquid chromatography [5]. The former has low analysis accuracy and cannot determine the content of single phenols; the latter has high analysis accuracy, but the analysis process is complicated, the speed is slow, and the sample is chemically polluted, which cannot meet the requirements of online component content monitoring during the production process. Near-infrared spectroscopy is a fast and non-destructive analysis technique, which can complete the detection of multi-components of substances within 1 min [6]. Near-infrared spectroscopy (NIR) is mainly the frequency-doubled and combined-frequency absorption spectrum of organic molecules. The frequency-doubled and combined-frequency absorption of the fundamental frequency vibrations of groups such as CH, NH, OH, and CO in material molecules are absorbed in the near infrared region, so Near-infrared technology is more suitable for the analysis of components that have a direct or indirect relationship with these groups [7]. Its outstanding advantages are fast analysis speed, high accuracy, and good stability. The polyphenols in sorghum grains contain a large number of functional groups such as OH, CH, OH, CH2, CHOH, and COROH, which have obvious absorption in the near infrared spectrum. The near-infrared mathematical model of polyphenols in sorghum grains can be established by chemometrics, which can quickly determine the content of polyphenols in sorghum grains and reduce the cost of detection. The near infrared prediction model of sorghum phenols has wide application value in high quality breeding and quality analysis of sorghum. This experiment used near infrared spectroscopy to quantitatively analyze the content of polyphenols in sorghum grains, and established total phenols, total flavonoids, condensed tannin, ferulic acid, protocatechuic aldehyde, anthocyanins and other phenols in sorghum. The mathematical model of substance content has higher analysis accuracy.
2 Experimental part
2.1 Collection and selection of experimental samples
The experimental materials were 33 sorghum varieties, 8 Sudan grass varieties and 19 sorghum Sudan grass hybrid varieties in Shanxi, Liaoning, Gansu, Xinjiang, Tianjin, Inner Mongolia and other places. The collected materials have a wide genetic source, and there are significant differences in tannin content among different materials and different parts of grains. Samples were taken from different parts of 60 test material seeds, including seed coat, hulled kernels, endosperm, embryo and whole seeds, to form 300 experimental samples. The sample was crushed by a pulverizer (Beijing Liuyi Factory), passed through a 0.18mm aperture sieve, and crushed a second time by a CyclotecTM 1093 cyclone mill (Foss), placed in a ziplock bag, kept at room temperature and protected from light, to be tested.
2.2 Determination of 6 phenolic substances in the sample
Reference [8], using Folin
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