ABSTRACT
Objective:To detect the colony number of bacteria, yeasts and molds in fermentation process of Pinelliae Rhizoma Fermentata (PRF), microbial flora species, and quantitatively analyze the dynamic changes of four dominant microorganisms at different fermentation time points of PRF, so as to provide experimental basis for exploring the processing mechanism of PRF. Method:According to Pharmaceutical Standard Preparation of Traditional Chinese Medicine Prescription of Ministry of Health of the People's Republic of China (the 10th volume), PRF was processed. The samples at five different fermentation time points (0, 30, 60, 90, 120 h) of PRF were taken, the culturing, isolation and purification of bacteria, yeasts and molds were carried out with selective media, and the colonies were counted. Fluorescence quantitative polymerase chain reaction (PCR) technique was employed to conduct absolute quantification of Bacillus subtilis, Paecilomyces variotii, Byssochlamys spectabilis and Aspergillus niger. The recombinant plasmids of these 4 microorganisms were used as the standard substances, and the standard curves were prepared after dilution of multiple ratios, quantitative analysis was performed on these 4 microorganisms in five samples at different processing time points (0, 30, 60, 90, 120 h) of PRF. Result:During the fermentation process of PRF, the number of bacteria was low with smooth change, while molds and yeasts grew dramatically at the late stage of fermentation and reached 1×106 CFU·mL-1 at the end of fermentation. At 5 different fermentation time points, the copy numbers of Bacillus subtilis were 3.53×105, 7.56×104, 1.58×105, 1.90×106, 1.85×106 copies·g-1, the copy numbers of Paecilomyces variotii were 0, 0, 0, 3.45×107, 4.15×108 copies·g-1, the copy numbers of Byssochlamys spectabilis were 0, 0, 0, 1.04×108, 2.28×108 copies·g-1, the copy numbers of Aspergillus niger were 0, 0, 9.48×105, 1.47×106, 7.56×106 copies·g-1, respectively. Conclusion:The change trend of microflora in the fermentation process of PRF can be reflected by the dynamic change of four dominant microorganisms, and molds may play an important role in the processing of PRF. Fluorescence quantitative PCR technique has the advantages of rapidity, sensitivity, good repeatability and high specificity, it is suitable for exploring processing mechanism of PRF.
ABSTRACT
Objective: To study the physiological and biochemical characteristics of four dominant microorganisms and the yellow pigment content of Pinelliae Rhizoma Fermentata (PRF), and provide basis for exploring the mechanism of PRF processing. Methods: The optimum growth temperature and pH value of the four dominant microorganisms Bacillus subtilis, Paecilomyces variotii, Byssochlamys spectabilis, and Aspergillus niger were studied. The ability of producing acidase, amylase, protease, and yellow pigment were determined. The yellow pigment content of each sample at different fermentation time points in process of PRF was determined. Results: The most suitable growth temperatures for B. subtilis, P. variotii, B. spectabilis, and A. niger were 35 ℃, 29 ℃, 29-31 ℃, and 39 ℃; And the optimum pH were 7.0, 7.0, 7.5, and 7.0, respectively. Four kinds of microorganisms had the ability to produce amylase and protease. P. variotii and B. spectabilis had the ability to produce yellow pigment. The content of yellow pigment were 69.875, 69.875, 71.750, 119.500, and 137.875 μg/g in the samples at different time points. Conclusion: Four kinds of dominant microorganisms may play an important role in fermentation process of PRF.
ABSTRACT
OBJECTIVE:To compare the composition changes of Aurantii fructus before and after fermentation processing and optimize its fermentation processing technology. METHODS:UPLC was conducted to compare the raw and fermentation processed products of same batch of Aurantii fructus,and ensure the chromatographic peaks after fermentation processing. Using peak areas of 4 chromatographic peaks and mildew characteristics of samples as index,fermentation temperature,humidity and time as factor, L9(34)orthogonal test was designed to optimize the fermentation processing technology,and verified it. RESULTS:After fermenta-tion processing,Aurantii fructus obviously showed 2 new monosaccharide glycosides components;the optimized fermentation tech-nology was as follows as fermentation temperature of 30 ℃,humidity of 70% and time of 7 d;verification test results showed RSD of each indicator of decoction pieces prepared by optimized fermentation technology in 3 tests were lower than 2.0%(n=3). CONCLUSIONS:Fermentation processing may lead obvious chemical composition changes in Aurantii fructus;the optimized fer-mentation processing technology can increase the contents of characteristic peaks.
ABSTRACT
To explore the processing mechanism of Aurantii Fructus decoction pieces used in Guangdong province and Hong Kong by analysing the chemical variation between raw and processed Aurantii Fructus with different methods based on UHPLC-Q-TOF-MS. The total ion chromatograms detected in positive and negative ion modes, and ion peak area ratio before and after processing were taken as variation indexes in the comparison. The results indicated that fermented Aurantii Fructus could produce three new ingredients, namely eriodictyol-7-glucoside, hesperetin-7-O-glucoside and 5-demethylnobiletin. At the same time, it could significantly increase the content of naringenin and hesperetin components, and could increase the content of such limonin derivatives as sudachinoid A, obacunoic acid and limoninand nomilinic acid. This suggests that the fermentation processing method of Aurantii Fructus decoction pieces used in Guangdong province and Hong Kong is of important significance for enhancing biological activity and bioavailability, and improving the clinical efficacy of Aurantii Fructus decoction pieces, and so is worth further protection and promotion.
ABSTRACT
Objective: To optimize the fermentation processing technology for fermented soybeans, and to identify that the secondary fermentation process is an important link in fermentation processing of fermented soybeans. Methods: According to Chinese Pharmacopoeia 2010, combined with the ancient way to produce fermented soybeans, the contents of total isoflavones, daidzein, and genistein were as chemical indexes, the finished product properties (color, smell, degree of shrinkage, cross section, and hardness) were as organoleptic quality indexes, and the processing parameters before and within the secondary fermentation process, including cooking time, fermentation temperature, fermentation time, secondary fermentation temperature and time were optimized. Results: The optimal processing technology was as follows: soybeans were boiled for 1.5 h after absorbing drug juice, and then fermented for 6-8 d at (30 ± 2)°C until yellow cladding. After washing off yellow cladding, soybeans were placed in the container, sealed using water, and put into secondary fermentation process. The container was in secondary fermentation for 12-15 d at (30 ± 2)°C. During the period of secondary fermentation, the soybeans were poured out every 3 d with stiring and slightly drying for four to five times, and at last was slightly steamed and dryed. The quality of fermented soybeans after secondary fermentation had more obvious advantage than that without secondary fermentation. The finished product had aromatic odor with light color and grain soft. The cross section color was brownish black and hand skin shrivel. The contents of total-isoflavones, daidzein, and genistein were at the highest value. Conclusion: The secondary fermentation process is an important link in fermentation processing of fermented soybeans and the key factor to affect the change of main chemical composition content and the finished product properties of fermented soybeans. The fermentation processing technology of fermented soybeans after optimization would lay the foundation of the regulation of production and the research of fermentation processing mechanism.