Flavor-enhanced codonopsis beer and its brewing method

Abstract

This invention relates to the field of food processing technology, and more particularly to a flavor-enhanced Codonopsis pilosula beer and its brewing method. The preparation method includes: malt saccharification, wort boiling, and fermentation; adding Codonopsis pilosula 20-40 minutes before the end of boiling, at a dosage of 0.3-1g per liter of wort; the fermentation includes: inoculating the wort with German wheat yeast and fermenting until the diacetyl content decreases to below 0.09 mg / L. This invention provides a Codonopsis pilosula German wheat beer, which, by introducing Codonopsis pilosula at a specific stage and cooperating with German wheat yeast fermentation, can maximize the harmony of the flavors of both, increase the ethyl acetate content, eliminate any abrupt medicinal taste, and simultaneously retain a large proportion of high-value nutrients such as atractylodes lactone III. This has significant application value in the field of traditional Chinese medicine beer.

Description

Technical Field

[0001] This invention relates to the field of food processing technology, and in particular to a flavor-enhanced Codonopsis pilosula beer and its brewing method. Background Technology

[0002] In recent years, with the increasing health awareness of consumers and the diversification and functionalization of the beer market, health-oriented beers that combine traditional Chinese medicine or food-medicine homologous ingredients with traditional beer brewing techniques have gradually become a research and development hotspot in the industry. Currently, various products with added ingredients such as goji berries, ginseng, and lingzhi mushrooms have appeared on the market. These products aim to retain the traditional flavor of beer while giving it health benefits, in order to meet the diverse needs of different consumer groups for beer quality and functionality.

[0003] In existing technologies, the introduction of Chinese herbal medicine ingredients into the beer brewing process typically includes the following methods: First, direct addition during the saccharification stage, where Chinese herbal medicine powder is added to the saccharification pot along with malt. This method can lead to the starch content of the herbs affecting the wort's sugar content, and some aroma components are lost during prolonged saccharification. Second, adding extracts during the fermentation stage, where Chinese herbal medicine extracts are added during the primary or secondary fermentation stages. While this method can retain some aroma, the extracts may introduce unwanted microorganisms and have poor compatibility with yeast metabolites, easily producing off-flavors. Third, adding in the boiling pot, where Chinese herbal medicines are added during the wort boiling stage. However, most existing solutions use a combination of multiple herbs without considering the synergy and compatibility with the fermentation characteristics and yeast flavor of wheat beer.

[0004] Therefore, existing technologies generally suffer from the following defects: poor flavor coordination and insufficient retention of active ingredients. Most food and medicine homologous beers have an overpowering herbal flavor after the addition of Chinese herbal ingredients, which masks the malt, hop, and yeast aromas of the beer itself, resulting in an uncoordinated taste and low consumer acceptance. Improper processing methods also lead to a significant loss of active ingredients and volatile aroma substances in Chinese herbal medicines. Summary of the Invention

[0005] To address the problems existing in the prior art, this invention provides a flavor-enhanced Codonopsis pilosula beer and its brewing method. By introducing Codonopsis pilosula at a specific stage and combining it with German wheat yeast fermentation, this invention maximizes the harmony of the flavors of both, eliminating any jarring medicinal taste, while also preserving a significant portion of high-value nutrients, resulting in a flavorful beer with both excellent flavor and active ingredients.

[0006] In a first aspect, the present invention provides a method for preparing beer, comprising: malt saccharification, wort boiling and fermentation, wherein Codonopsis pilosula is added 20 to 40 minutes before the end of the boiling process, and the amount of Codonopsis pilosula added is 0.3 to 1 g per liter of wort. The fermentation process includes inoculating the wort with German wheat yeast and fermenting until the diacetyl content is reduced to below 0.09 mg / L.

[0007] Furthermore, fermentation can be carried out until the content of atractylodes lactone III is above 8 μg / L and the content of ethyl acetate is increased to above 30 mg / L.

[0008] Existing medicinal beer products, after adding traditional Chinese medicine ingredients, have an overpowering herbal flavor that masks the beer's natural malt, hop, and yeast aromas, resulting in an unbalanced taste and low consumer acceptance. This invention has identified two key reasons for this: firstly, adding the herbs during the saccharification stage or prolonged boiling leads to a significant loss of active ingredients (such as ginsenosides, polysaccharides, and saponins in Codonopsis pilosula) and volatile aroma compounds; secondly, existing technologies do not consider synergistic design with specific yeast flavors. Therefore, the Codonopsis pilosula beer provided by this invention adds Codonopsis pilosula 20-40 minutes before the end of boiling, and limits the specific amount of Codonopsis pilosula used. This maximizes the preservation of the active ingredients and volatile aroma compounds of Codonopsis pilosula, while allowing its subtle sweetness to complement the aforementioned yeast flavors. Furthermore, the reducing sugars in Codonopsis pilosula can slightly participate in the Maillard reaction, increasing the beer's roundness. The resulting Codonopsis pilosula beer has no jarring herbal taste and retains a large proportion of high-value nutrients, possessing both excellent flavor and the active ingredients of Codonopsis pilosula, making it of significant application value.

[0009] Adding Codonopsis pilosula for more than 40 minutes or less than 20 minutes will result in a significant loss of volatile aroma compounds and active substances in the Codonopsis pilosula, or prevent their full release, which will further disrupt the subsequent fermentation system. The resulting beer will have an abrupt medicinal taste and an uncoordinated flavor, while the content of active substances in Codonopsis pilosula will be limited.

[0010] Furthermore, add Codonopsis pilosula 25-35 minutes before the end of the boiling process, at a dosage of 0.4-0.6g per liter of wort.

[0011] Furthermore, the German wheat yeast is selected from one or more of Saccharomyces cerevisiae WB-06, Saccharomyces cerevisiae WLP300, or Saccharomyces cerevisiae WY3068; Preferably, the German wheat yeast is brewer's yeast WB-06.

[0012] Furthermore, when the German wheat yeast is *Saccharomyces cerevisiae* WB-06, the inoculum size is 1.5 × 10⁻⁶. 5~8 cells / mL; the fermentation conditions include: Ferment at 18-20℃ for 15-25 days.

[0013] In this invention, brewing yeast WB-06 was fermented in the presence of Codonopsis pilosula. Besides producing typical banana (isoamyl acetate) and clove (4-vinylguaiacol) flavors, an unknown sesquiterpene compound (retention index approximately 1520, characteristic ion m / z 161, 105, 91) was detected by two-dimensional gas chromatography-tandem mass spectrometry (GC×GC-MS / MS). This compound was not detected in ordinary German wheat beer or Codonopsis pilosula aqueous extract. This indicates that certain precursors in Codonopsis pilosula (such as glycosides) are hydrolyzed by yeast enzymes during fermentation and converted into new volatile terpenes, thus imparting a unique, delicate herbal aroma to Codonopsis pilosula beer—a flavor component not found in WB-06 fermentation alone or in Codonopsis pilosula extract alone.

[0014] Furthermore, BU analysis revealed that Codonopsis pilosula beer has a higher concentration of bitter substances compared to regular beer. However, when using an electronic tongue to simulate human tasting, the perceived bitterness of Codonopsis pilosula beer was actually lower than that of regular beer. This may be because the polysaccharides and sweet amino acids (such as glycine and alanine) in Codonopsis pilosula mask or antagonize bitterness. On the one hand, they physically cover bitter taste receptors; on the other hand, they suppress bitterness perception by enhancing the richness of sweetness and umami (the electronic tongue showed that the sweetness SCS of Codonopsis pilosula beer was 7, higher than 5.4 in regular beer, and the umami richness CPS was 6.9, higher than 5.5). Therefore, the Codonopsis pilosula beer prepared by the method provided in this invention achieves a unique "low bitterness and sweet aftertaste" taste, which is completely consistent with the feedback from human sensory evaluation that "Codonopsis pilosula beer has a lower bitterness."

[0015] Furthermore, this invention discovered that ordinary beer is high in typical umami amino acids (such as glutamic acid). In Codonopsis beer, these substances are transformed into other components, making the taste complex composed of sweetness, umami (even at low levels), peptides, polysaccharides, etc., more complex and harmonious, resulting in a "rich and layered" taste.

[0016] Furthermore, bitter flowers are added 8-15 minutes after the wort begins to boil, and fragrant flowers are added 8-15 minutes before the boiling ends. Preferably, the bitter hops are Saaz hops, and the amount added is 0.2~0.4g per liter of wort; The aromatic hops are Simco hops, added at a rate of 0.1-0.3g per liter of wort.

[0017] Existing beer products containing both food and medicinal ingredients often suffer from poor bitterness control. Furthermore, Codonopsis pilosula itself contains trace amounts of saponin bitterness, which, if not properly processed, can produce an unpleasant bitterness, affecting the overall taste of the beer. Therefore, this invention further employs specific bitter and aromatic flower treatments to further harmonize the overall flavor and promote the expression of aroma compounds.

[0018] Further, the maltization includes: After mixing malt and water at a mass ratio of 1:(3~5), the mixture was treated at 65~72℃ for 50~70 minutes, and then filtered.

[0019] Further, by weight, the malt comprises: 50-70 parts wheat malt, 30-50 parts barley malt, and 3-10 parts roasted malt; and / or, The filtration is carried out at a temperature of 75~80℃.

[0020] This invention employs a specific ratio of malt blend (50-70 parts wheat malt, 30-50 parts barley malt, and 3-10 parts caramel malt) and a single-step infusion saccharification method (68℃, 60 min). The resulting wort contains approximately 12-14°P of fermentable sugars (mainly maltose and glucose) and approximately 150-200 mg / L of free amino nitrogen (FAN). The addition of Codonopsis pilosula further provides polysaccharides and trace elements (such as Zn). 2+ Mg 2+ The experiment found that, compared with the control group which only used maltose without Codonopsis pilosula, the Codonopsis pilosula group showed an approximately 15% increase in sugar consumption rate during the initial fermentation stage (0-48h), a shortened fermentation cycle of approximately 2 days (20 days vs. 22 days), and a 58% increase in final ethyl acetate production. Simultaneously, GC-MS analysis showed that the total ester content in Codonopsis pilosula beer (152.3 mg / L) was significantly higher than that in the control group (98.7 mg / L). This indicates that the nutrients in Codonopsis pilosula and the maltose products formed a better yeast nutritional balance, offsetting the possible slight inhibitory effect of Codonopsis pilosula saponins on yeast and promoting yeast growth and ester metabolism.

[0021] Furthermore, it also includes: post-ripening; The post-fermentation process includes: after fermentation, cooling the temperature to 0-6℃ at a rate of 0.4-0.8℃ / h, and performing post-fermentation treatment for 5-8 days under CO2 saturation.

[0022] Secondly, the present invention provides a beer prepared by the aforementioned preparation method; Preferably, it also includes fruit; More preferably, the fruit includes one or more of plums, citrus fruits, strawberries, or grapes.

[0023] Thirdly, the present invention provides a beer, the raw materials of which include: Codonopsis pilosula, brewing yeast WB-06, wheat malt, barley malt, caramel malt, Saaz hops and Simco hops; Preferably, by weight, the raw materials include: 3-10 parts of Codonopsis pilosula, 800-1600 parts of wheat malt, 400-1200 parts of barley malt, 80-120 parts of roasted malt, 2-4 parts of Saaz hops and 1-3 parts of Simco hops.

[0024] The present invention has the following beneficial effects: This invention provides a Codonopsis pilosula German-style wheat beer. By introducing Codonopsis pilosula at a specific stage and combining it with German wheat yeast fermentation, the loss of active ingredients and flavor substances can be prevented to the greatest extent. At the same time, the flavors of the two are coordinated, resulting in a flavor beer without any abrupt medicinal taste, with enhanced flavor (ethyl acetate content significantly increased, for example, by more than 50%), while retaining most of the active ingredients of Codonopsis pilosula (such as atractylodes lactone III ≥ 8.0 μg / L), which has high economic value. Attached Figure Description

[0025] To more clearly illustrate the technical solutions in this invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0026] Figure 1 This is a brewing process diagram of Codonopsis pilosula German-style wheat beer provided in an embodiment of the present invention.

[0027] Figure 2 The results are obtained from the electronic tongue detection of Codonopsis pilosula beer, ordinary German wheat, and Codonopsis pilosula extract provided in this embodiment of the invention.

[0028] Figure 3 The results of electronic tongue principal component analysis of Codonopsis pilosula beer, ordinary German wheat, and Codonopsis pilosula extract provided in the embodiments of the present invention are shown.

[0029] Figure 4 The results of electronic tongue discriminant factor analysis of Codonopsis pilosula beer, ordinary German wheat, and Codonopsis pilosula extract provided in the embodiments of the present invention are shown.

[0030] Figure 5 This is the first part of the volcano diagram results of the volatile flavor compounds of Codonopsis pilosula beer and ordinary beer provided in the embodiments of the present invention.

[0031] Figure 6 This is the second part of the volcano diagram results of the volatile flavor compounds of Codonopsis pilosula beer and regular beer provided in the embodiments of the present invention.

[0032] Figure 7 This is the first part of the volcano diagram results of the volatile flavor compounds of Codonopsis pilosula extract and ordinary beer provided in the embodiments of the present invention.

[0033] Figure 8 This is the second part of the volcano diagram results of the volatile flavor compounds of Codonopsis pilosula extract and ordinary beer provided in the embodiments of the present invention.

[0034] Figure 9 These are the main component analysis results of volatile flavor substances in Codonopsis beer, ordinary German wheat, and Codonopsis extract provided in the embodiments of the present invention.

[0035] Figure 10 The partial least squares discriminant analysis results of volatile flavor compounds in Codonopsis beer, ordinary German wheat, and Codonopsis extract provided in the embodiments of the present invention are as follows.

[0036] Figure 11 These are the PLS-DA variable importance results for volatile flavor compounds in Codonopsis beer, ordinary German wheat, and Codonopsis extract provided in the embodiments of the present invention.

[0037] Figure 12 The results are heatmap clustering analysis of volatile flavor compounds in Codonopsis pilosula beer, ordinary German wheat, and Codonopsis pilosula extract provided in the embodiments of the present invention. Detailed Implementation

[0038] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of this invention, not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0039] Unless otherwise specified, the experimental methods involved in the following embodiments are conventional methods in the art. For example, you can refer to the experimental manual in the art or follow the conditions recommended in the manufacturer's instructions.

[0040] Unless otherwise specified, all experimental materials and reagents used in the following examples are commercially available.

[0041] The raw materials used in the following examples (barley malt, wheat malt, caramel malt, Czech Saaz hops, American Simco hops, WB06 yeast, etc.) can all be purchased commercially (e.g., ordered from Weifang Lanqiao Craft Beer Raw Material Factory).

[0042] Example 1 I. Brewing process of Codonopsis pilosula German-style wheat beer (process as follows) Figure 1 (As shown).

[0043] 1. Raw material preparation.

[0044] Malt: 60% wheat malt, 40% barley malt, plus 5% roasted malt.

[0045] Water: Soft water that meets drinking water standards.

[0046] Hops: Saz hops (bitter hops), Simco hops (fragrant hops).

[0047] Yeast: German Ale WB06 yeast (brewing yeast).

[0048] Codonopsis pilosula: Three-year-old Codonopsis pilosula from Wushan, sliced ​​and dried for later use.

[0049] 2. Saccharification and filtration.

[0050] The single-step saccharification method was adopted: pulverized malt (12 kg of wheat malt, 8 kg of barley malt, and 1 kg of caramel malt, totaling 21 kg) was mixed with water at a ratio of 1:3.5, 73.5 kg of water was added, and the temperature was raised to 68℃ and kept at that temperature for 60 min to complete starch saccharification.

[0051] The mixture was heated to 78°C and filtered to separate approximately 100 L of wort and lees.

[0052] 3. Boil and add Codonopsis pilosula.

[0053] Boil 100 L of the wort for 90 minutes.

[0054] Add Czech Saz hops (bitter hops, 0.3 g / L, or 30 g) 10 minutes after boiling begins.

[0055] Add the Codonopsis pilosula slices 30 minutes before the end of boiling (dosage: 0.5 g / L wort, i.e., 50 g).

[0056] Add American Simco hops (fragrant hops, 0.2 g / L, or 20 g) 10 minutes before the end of boiling.

[0057] After boiling, transfer to a whirlpool and let stand for 20 minutes to remove heat-coagulated material.

[0058] 4. Cooling and fermentation.

[0059] The wort is cooled to 18°C ​​using a plate heat exchanger.

[0060] After oxygenation, inoculate with WB06 yeast (inoculation amount 1.5 × 10⁶). 7 (cells / mL).

[0061] The primary fermentation temperature was controlled at 18-20℃, and the fermentation time was 20 days. During this period, sugar content, diacetyl, and yeast count were monitored daily.

[0062] Cooling begins when the diacetyl content drops below 0.09 mg / L.

[0063] 5. Post-ripening and aging.

[0064] After the primary fermentation is completed, the temperature is lowered to 4℃ at a rate of 0.5℃ / h, and then allowed to ripen for 7 days.

[0065] During the post-ripening period, maintain the tank pressure at 0.15 MPa to saturate it with carbon dioxide.

[0066] 6. Packaging.

[0067] Bottled products are produced using an aseptic filling line.

[0068] The finished beer is naturally cloudy (containing active yeast) and has a foam retention of ≥150 seconds.

[0069] 2. Codonopsis and Plum Beer.

[0070] Based on the Codonopsis pilosula beer, Wushan crisp plum puree (10% v / v) is added during the post-fermentation stage, and secondary fermentation is carried out for 5-7 days to obtain a flavor beer with a blend of fruit and ginseng aroma.

[0071] Experimental Example 1 This experimental example focuses on the Codonopsis pilosula beer obtained in Example 1. The contents of diacetyl, atractylodes lactone III, and ethyl acetate were detected by liquid chromatography-tandem mass spectrometry; the color and bitterness were detected by spectrophotometer; and the alcohol content and original wort concentration were detected by densitometer.

[0072] Using standard German wheat beer as a control (compared to the brewing method provided in Example 1, only without the addition of Codonopsis pilosula, all other components and processes are the same; also the subsequent standard beer and standard blank beer), the results are shown in the table below: Table 1. Results of characteristic tests on Codonopsis pilosula beer

[0073] The results above show that the content of ethyl acetate in Codonopsis beer is significantly higher than that without Codonopsis, which indicates that the addition of Codonopsis effectively increases the content of aroma and flavor substances in beer.

[0074] Experiment Example 2 This experimental example further uses GC×GC-MS / MS, electronic nose, and electronic tongue to detect the Codonopsis pilosula beer obtained in Example 1, and the following results are obtained: The results of the electronic tongue are shown in Table 2 and Figures 2-4 As shown, Y1 = regular beer, Y2 = Codonopsis pilosula beer, and Y3 = Codonopsis pilosula extract.

[0075] Table 2 Electronic tongue detection results

[0076] Figure 3 The principal component analysis (PCA) results of the electronic tongue shown indicate that: The cumulative variance contribution rate of PC1 and PC2 reached 96.65%, with a discrimination index of 87, indicating excellent sample differentiation performance. The figure shows that parallel sample points of the same sample are closely clustered, indicating good experimental repeatability and stable and reliable data. Sample points from different treatment groups are distributed in different regions, with the distance between the Codonopsis pilosula beer sample and the regular beer sample being relatively large, indicating a significant difference in their overall taste characteristics. The three groups of samples are orderly distributed along the PC1 axis. The taste characteristics of Codonopsis pilosula beer are between those of regular beer and Codonopsis pilosula extract, proving that during the brewing process, the bitterness, astringency, and aftertaste of Codonopsis pilosula are effectively transferred to the beer system, achieving flavor fusion.

[0077] To further distinguish the taste characteristics of different samples, discriminant factor analysis (DFA) was used to classify the data. Figure 4 DF1 contributed 73.434%, DF2 contributed 26.566%, and their combined contribution was 100%, which can fully characterize the overall taste differences among the three groups of samples.

[0078] The three groups of samples were completely independent and separated without any overlap. Y1 (plain blank beer, red): concentrated in the left area; Y2 (Codonopsis pilosula beer, blue): Located on the far right; Y3 (Codonopsis pilosula extract, green): Distributed in the upper area.

[0079] The close clustering of parallel samples within each group indicates high stability of taste perception and excellent experimental repeatability.

[0080] Discriminant factor analysis (DFA) results showed that DF1 and DF2 could fully explain the taste differences between samples. Ordinary blank beer, Codonopsis beer, and Codonopsis extract could be completely distinguished by the electronic tongue. The samples within the group showed high aggregation and the separation effect between the groups was significant, proving that the taste characteristics of Codonopsis extract can be effectively integrated into beer, giving Codonopsis beer a unique taste profile that is different from ordinary beer and pure Codonopsis extract.

[0081] This demonstrates that the electronic tongue can accurately distinguish between ordinary blank beer, Codonopsis beer, and Codonopsis extract, with good data repeatability and high reliability. The taste of Codonopsis beer is not simply an additive combination, but rather forms a new and stable flavor between pure beer and pure Codonopsis extract, achieving a synergistic flavor between Codonopsis and German wheat beer.

[0082] Bitterness (PKS): Codonopsis beer (5.9) was significantly lower than control beer (6.3). Combined with chemical bitter substances (BU: Codonopsis beer 11.1 vs control 10.25), it indicates that the polysaccharides and sweet amino acids in Codonopsis effectively masked the bitterness, achieving "low bitterness and sweet aftertaste".

[0083] Differences in Umami (NMS) and Umami Richness (CPS): Notably, the control beer had a high NMS of 8.5, significantly higher than the Codonopsis beer's 3.9; however, the Codonopsis beer's CPS was 6.9, significantly higher than the control's 5.5. This is likely because the NMS sensor primarily responds to typical umami amino acids such as glutamic acid and aspartic acid. In regular beer, yeast metabolism produces large amounts of these amino acids, hence the high NMS value. In Codonopsis beer, these amino acids may be reduced due to their participation in the Maillard reaction (with Codonopsis sugars) or their use by yeast in other metabolic pathways.

[0084] The CPS sensor (Umami Richness, also known as the "Koku" sensor) is a comprehensive response to the complexity and harmony of taste. It is sensitive to small molecule peptides, organic acids, sweet substances, and the complex effects of umami and sweet / bitter tastes. Codonopsis pilosula is rich in polysaccharides, small molecule peptides, and sweet amino acids (such as glycine and alanine). During fermentation: ① Polysaccharides degrade into oligosaccharides, increasing the body of the beer; ② Codonopsis pilosula protein hydrolysis produces small molecule peptides, directly increasing the CPS value; ③ The high sweetness of Codonopsis pilosula beer (SCS 7.0 vs 5.4) synergistically with residual umami substances to form a more complex and harmonious taste complex. Therefore, although the concentration of typical umami amino acids decreases, the "richness" and "fullness" of the taste are actually enhanced.

[0085] This data perfectly corresponds to the sensory evaluation description of "smooth and mellow with a distinct aftertaste," proving that Codonopsis beer is not just a single, intense umami flavor, but rather a more sophisticated and complex mellow taste created through the fermentation and transformation of Codonopsis.

[0086] Experimental Example 3 The present invention further analyzed the ordinary German wheat (sample 1), the Codonopsis pilosula beer (sample 2) prepared in Example 1, and the Codonopsis pilosula extract (sample 3) by GC×GC-MS / MS.

[0087] 1. Overview of overall differences.

[0088] Table 3 Overview of volatile components in samples 1-3

[0089] 2. Analysis of differences in unique components.

[0090] (1) Unique components of sample 1 (147 kinds).

[0091] Typical examples: Esters: such as 1,3-propanediol diacetate and dibutyl phthalate.

[0092] Alcohols: such as 1,2-dihydrolinalool, 2-nonanol, and 2-octanol.

[0093] Alkanes: such as pentadecane.

[0094] Silanes / siloxanes: such as methyltriacetoxysilane and decamethylpentasiloxane.

[0095] Characteristics: Mostly esters, alcohols, and siloxanes produced during beer fermentation, with some being flavor compounds or packaging migrations.

[0096] (2) Unique components of sample 2 (130 kinds).

[0097] Typical examples: Alcohols: such as 2-decyl alcohol.

[0098] Esters: such as ethyl butyrate.

[0099] Alkenes, ketones, and phenols: such as (E)-β-farnesene, α-juniperol, and p-tert-butylphenol.

[0100] Features: Contains some terpenes and phenols derived from Codonopsis pilosula, while retaining characteristic components of beer fermentation.

[0101] (3) Unique components of sample 3 (Codonopsis pilosula extract) (165 kinds).

[0102] Typical examples: Alcohols: such as 1-nonanol, 1-pentanol, and isoborneol.

[0103] Ketones: such as 2-nonanone and 2-octanone.

[0104] Aldehydes: such as heptanal and pentanal.

[0105] Acids: such as nonanoic acid and ethyl nonanoate.

[0106] Terpenes: such as camphor, linalool, and limonene.

[0107] Features: It has the richest composition, containing terpenes, alcohols, ketones, and aldehydes from natural plant sources, which are highly correlated with the medicinal components of Codonopsis pilosula.

[0108] (4) There are a total of 69 species in samples 1 and 2.

[0109] (5) There are a total of 17 species in samples 1 and 3.

[0110] (6) Codonopsis pilosula related characteristic components (shared by samples 2 and 3, absent in sample 1).

[0111] Table 4. Common characteristic components of samples 2 and 3

[0112] These components were not detected in regular beer, indicating that they are closely related to the addition of Codonopsis pilosula or the fermentation process.

[0113] in conclusion: Sample 1 (regular beer) has a relatively simple composition, mainly consisting of fermentation byproducts and ester flavor compounds.

[0114] Sample 2 (Codonopsis pilosula fermented beer) While retaining the characteristic components of beer, it introduced some terpenes, phenols and other compounds derived from Codonopsis pilosula.

[0115] Sample 3 (Codonopsis pilosula extract) has the most complex composition and is rich in natural plant metabolites, which is the main source of Codonopsis pilosula's active ingredients.

[0116] Thirteen characteristic components of Codonopsis pilosula are important markers for distinguishing whether Codonopsis pilosula has been added.

[0117] 3. Volcano map.

[0118] (1) Codonopsis beer and regular beer.

[0119] The results are as follows Figure 5 and Figure 6 As shown, a total of 165 compounds were identified (18+13+134), of which 13 were significantly upregulated (red), 18 were significantly downregulated (blue), and 134 showed no significant difference (gray).

[0120] The overall changes were mild, with post-fermentation differences concentrated in a few key compounds.

[0121] Approximately 81% of the compounds (134 / 165) showed no significant difference between Codonopsis beer and regular beer (|log2FC|<1 and p>0.05), indicating that Codonopsis beer still maintains the typical flavor structure of German wheat beer.

[0122] (2) Codonopsis pilosula extract VS regular beer.

[0123] The results are as follows Figure 7 and Figure 8 As shown in the results, there are 52 cases with significant upregulation (red), 36 cases with significant downregulation (blue), and 91 cases with no significant difference.

[0124] Table 5. Differences in volatile substances among different comparison groups

[0125] The results from different comparison groups show that, before fermentation, there are significant differences in the volatile component profiles of Codonopsis pilosula extract and beer. The compounds significantly upregulated in the Codonopsis pilosula extract (log2FC>1, p<0.05) are mainly characteristic terpenes of Codonopsis pilosula (such as atractylodes lactone III, atractylone, β-caryophyllene, etc.) and some aldehydes and furans. The compounds significantly downregulated are mainly beer-specific esters, higher alcohols, and hop-derived terpenes. The absolute value of log2FC can reach as high as 6.0 (corresponding to a fold change of 64 times), and the p value is extremely low (-log10p reaches as high as 5.8). This indicates that the volatile component profile of Codonopsis pilosula extract itself is fundamentally different from that of the beer matrix, and direct mixing would lead to flavor conflicts.

[0126] In contrast to the above results, after fermentation, approximately 80% of the volatile components in Codonopsis pilosula beer and regular beer showed no significant difference. Specific differences were observed in compounds such as ethyl acetate and atractylodes lactone III. This result indicates that by adding Codonopsis pilosula before the end of boiling and fermenting with German wheat yeast as described in this invention, the volatile components in Codonopsis pilosula that were originally incompatible with beer were effectively transformed or metabolized, resulting in a final product with a volatile component profile highly similar to regular German wheat beer, without introducing any abrupt medicinal flavor. Simultaneously, the non-volatile components (polysaccharides, sweet amino acids, etc.) brought by Codonopsis pilosula significantly improved the beer's taste quality (low bitterness, sweet aftertaste, and full-bodied flavor).

[0127] 4. Principal component analysis (PCA).

[0128] The present invention further performed principal component analysis (PCA) on the peak areas of all obtained compounds, and the results are shown in the figure. Among them, the contribution rate of the first principal component (PC1) is 51.8%, the contribution rate of the second principal component (PC2) is 12.8%, and the cumulative contribution rate reaches 64.6%, which can effectively characterize the overall volatility differences among samples.

[0129] like Figure 9 As shown, the Codonopsis pilosula extract (sample 3) is located in the positive region of PC1, showing significant separation from the two beer samples in the PC1 direction, indicating that the volatile component spectrum of the Codonopsis pilosula extract itself is fundamentally different from that of the beer matrix. Both regular beer (sample 1) and Codonopsis pilosula fermented beer (sample 2) are located in the negative region of PC1, and are relatively close to each other, verifying the conclusion that their flavor skeletons are similar. Furthermore, further observation in the PC2 direction reveals a slight separation between the Codonopsis pilosula fermented beer and regular beer, presumably due to the introduction of characteristic terpenoid components of Codonopsis pilosula (such as atractylodes lactone III) and changes in the content of some esters.

[0130] In summary, the PCA results show that there are significant differences between Codonopsis pilosula extract and beer matrix. However, after adding Codonopsis pilosula before the end of boiling and fermenting with German wheat yeast as described in this invention, the overall profile of volatile components in Codonopsis pilosula beer is highly similar to that of ordinary beer, successfully achieving a harmonious integration of Codonopsis pilosula and beer flavor and avoiding an abrupt medicinal taste.

[0131] 5. Partial Least Squares Discriminant Analysis (PLS-DA).

[0132] like Figure 10 As shown, this invention establishes a PLS-DA model and performs leave-one-out cross-validation. The results are shown in the Partial Least Squares Discriminant Analysis (PLS-DA). When using two principal components, the model accuracy reaches 100%, R0 2 =1.00, Q 2 =0.90; Q = 0.90 when there are 3 principal components 2 =0.95. Two principal components are sufficient to perfectly distinguish the three types of samples, and the predictive ability is excellent. This further verifies the aforementioned conclusion.

[0133] 6. PLS-DA Variable Importance (VIP) Plot.

[0134] like Figure 11 As shown, the VIP (Variable Importance in Projection) value reflects the degree of contribution of each variable to sample classification in the PLS-DA model. Generally, variables with VIP>1 are considered to have a significant impact on distinguishing different samples.

[0135] Only Ethyl 9-decenoate had a VIP value greater than 1.5, while the VIP values ​​of the remaining compounds were all less than 1. This result indicates that at the volatile component level, there is no significant difference in VIP values ​​between Codonopsis pilosula beer and regular beer, meaning their volatile component profiles are similar. The high-VIP Ethyl 9-decenoate and other compounds with VIP values ​​close to 1 (such as 2-methoxy-4-vinylphenol, decanoic acid, and octanoic acid) primarily contribute to distinguishing the Codonopsis pilosula extract from the two types of beer, reflecting the essential differences between the extract and the beer matrix. This result is corroborated by PCA and volcano plot analysis, demonstrating that the fermentation process described in this invention successfully eliminates significant incompatible components in the Codonopsis pilosula extract, making the final product's volatile component profile converge with that of regular German wheat beer, achieving a harmonious flavor fusion.

[0136] 7. Heatmap clustering analysis.

[0137] like Figure 12As shown, this invention performs Z-score normalization on all detected volatile components, generates heatmaps, and clusters them. Sample clustering shows that regular beer and Codonopsis pilosula beer initially cluster together, followed by Codonopsis pilosula extract. Component clustering shows that beer-characteristic components such as esters, higher alcohols, and phenols are highly expressed in both types of beer but lowly expressed in Codonopsis pilosula extract; characteristic terpenes of Codonopsis pilosula are highly expressed in Codonopsis pilosula extract but lowly expressed in both types of beer (among which atractylodes lactone III was detectable in Codonopsis pilosula beer but not in regular beer). The heatmaps visually confirm the elimination effect of fermentation on incompatible volatile components and the high similarity in flavor skeleton between Codonopsis pilosula beer and regular beer.

[0138] The above multivariate statistical analysis results consistently show that there are significant differences in the volatile components between Codonopsis pilosula extract and the beer matrix. However, after adopting the process described in this invention (adding Codonopsis pilosula and German wheat yeast for fermentation before the end of boiling), the volatile component profile of Codonopsis pilosula beer is basically consistent with that of ordinary German wheat beer, successfully achieving a harmonious integration of flavors while retaining characteristic markers of Codonopsis pilosula such as atractylodes lactone III. This invention provides an effective means to solve the technical problem of flavor incompatibility in beer made from food and medicine homologous ingredients.

[0139] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing beer, comprising: Malt saccharification, wort boiling and fermentation, characterized in that Codonopsis pilosula is added 20 to 40 minutes before the end of the boiling process, and the amount of Codonopsis pilosula added is 0.3 to 1 g per liter of wort; The fermentation process includes inoculating the wort with German wheat yeast and fermenting until the diacetyl content is reduced to below 0.09 mg / L.

2. The preparation method according to claim 1, characterized in that, Add Codonopsis pilosula 25-35 minutes before the end of the boiling process, at a rate of 0.4-0.6g per liter of wort.

3. The preparation method according to claim 1 or 2, characterized in that, The German wheat yeast is selected from one or more of brewer's yeast WB-06, brewer's yeast WLP300 or brewer's yeast WY3068; Preferably, the German wheat yeast is brewer's yeast WB-06.

4. The preparation method according to claim 3, characterized in that, When the German wheat yeast is Saccharomyces cerevisiae WB-06, the inoculum size is 1.5 × 10⁻⁶. 5~8 cells / mL; the fermentation conditions include: Ferment at 18-20℃ for 15-25 days.

5. The preparation method according to any one of claims 1-4, characterized in that, Add bitter flowers 8-15 minutes after the wort begins to boil, and add fragrant flowers 8-15 minutes before the end of the boiling process. Preferably, the bitter hops are Saaz hops, and the amount added is 0.2~0.4g per liter of wort; The aromatic hops are Simco hops, added at a rate of 0.1-0.3g per liter of wort.

6. The preparation method according to any one of claims 1-5, characterized in that, The malt saccharification includes: After mixing malt and water at a mass ratio of 1:(3~5), the mixture was treated at 65~72℃ for 50~70 minutes, and then filtered.

7. The preparation method according to claim 6, characterized in that, By weight, the malt comprises: 50-70 parts wheat malt, 30-50 parts barley malt, and 3-10 parts roasted malt; and / or, The filtration is carried out at a temperature of 75~80℃.

8. The preparation method according to any one of claims 1-7, characterized in that, Also includes: Post-ripening; The post-fermentation process includes: after fermentation, cooling the temperature to 0-6℃ at a rate of 0.4-0.8℃ / h, and performing post-fermentation treatment for 5-8 days under CO2 saturation.

9. A beer, characterized in that, Prepared by the preparation method according to any one of claims 1-8; Preferably, it also includes fruit; More preferably, the fruit includes one or more of plums, citrus fruits, strawberries, or grapes.

10. A beer, characterized in that, The ingredients include: Codonopsis pilosula, brewing yeast WB-06, wheat malt, barley malt, roasted malt, Saaz hops and Simco hops; Preferably, by weight, the raw materials include: 3-10 parts of Codonopsis pilosula, 800-1600 parts of wheat malt, 400-1200 parts of barley malt, 80-120 parts of roasted malt, 2-4 parts of Saaz hops and 1-3 parts of Simco hops.

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