CN112094714A

CN112094714A - Preparation method of mulberry wine

Info

Publication number: CN112094714A
Application number: CN202011059700.XA
Authority: CN
Inventors: 殷浩; 王香君; 吴劲轩; 夏川林; 蒲军; 夏文银
Original assignee: Sericultural Research Institute of Sichuan Academy of Agricultural Sciences
Current assignee: Sericultural Research Institute of Sichuan Academy of Agricultural Sciences
Priority date: 2020-09-30
Filing date: 2020-09-30
Publication date: 2020-12-18

Abstract

The invention relates to the technical field of wine brewing, in particular to a preparation method of mulberry wine. The method adopts whole fruit fermentation process, and the mulberry fruit wine or mulberry brandy prepared by the method has high quality, good sense and high yield. Wherein, the yield of the mulberry fruit wine is 50 percent to 55 percent, and the yield of the brandy is 1 ton of pomace, so that 50L of mulberry brandy can be obtained.

Description

Preparation method of mulberry wine

Technical Field

The invention relates to the technical field of wine brewing, in particular to a preparation method of mulberry wine.

Background

Mulberry is the fruit cluster of Morus alba L of Moraceae. Also named mulberry, sugarcane, mulberry, etc. The fruit is sweet and has much juice, and is one of the fruits which are often eaten by people. The mulberry is harvested when the fruits are ripe in 4-6 months every year, impurities are removed, and the mulberry is dried in the sun or is slightly steamed and then dried in the sun for eating.

Mulberries are rich in a variety of nutrients required by the human body, including amino acids, vitamins and minerals, as well as a variety of active substances. The main components contained in Mori fructus include water, saccharide, protein, lipid, free acid, crude fiber, etc., and contain 6 vitamins such as Vc, Va, sulfa-tin, riboflavin, etc., 19 amino acids (wherein the essential amino acids are 7), and abundant microelements and mineral components.

The mulberry belongs to berry fruits, contains a large amount of reducing sugar and a large amount of anthocyanin, under the action of yeast, the reducing sugar is used as a carbon source, protein is used as a nitrogen source, the reducing sugar in the mulberry juice is oxidized into ethanol and carbon dioxide through a series of metabolic reactions, and the generation process of alcohols, esters, acids and the like is accompanied in the microbial metabolism process. With the improvement of the living standard and the living quality of people in China, the drinking concept and the consumption habit of people are greatly changed. In view of health and nutrition, more people tend to select the fruit wine with rich nutrition and mild and low-degree fermentation, and the mulberry fruit wine is popular with more and more people in recent years as a new variety of fruit wine.

The mulberry fruit has a low density and a low juice yield, and thus has a high demand for a brewing process. In the prior mulberry wine brewing process, various problems still exist, such as the obtained fruit wine has high total acid content, insufficient anthocyanin content, unstable wine body turbidity and the like. These problems reduce the sensory quality of mulberry wine and limit the promotion and development of mulberry wine.

Disclosure of Invention

In view of the above, the technical problem to be solved by the present invention is to provide a method for preparing mulberry wine with good quality, which can obtain mulberry wine or mulberry brandy with high yield.

The preparation method of the mulberry fruit wine provided by the invention comprises the following steps:

crushing: crushing mulberry fruits with reducing sugar content not lower than 8g/100g, and adding 135-150 g of cane sugar to each kg of mulberry fruits to obtain fermentation liquor;

enzymolysis: after the fermentation liquor is sulfurized, carrying out enzymolysis by using pectinase, wherein the addition amount of the pectinase is 20 mg/kg;

main fermentation: controlling the temperature to be 20-25 ℃, adding 0.1-0.3 g/kg of yeast and 0.1-0.3 g/kg of fermentation aid, pressing a wine cap every 12 hours, and circularly stirring for 20-40 min; removing wine caps after 24-48 h, adding cane sugar, continuing fermenting until the alcoholic strength reaches 12% vol and the total sugar content is less than or equal to 1%, and taking supernate;

and (3) after-fermentation: and sealing and standing the supernatant at 15-20 ℃ for 1 month, taking the supernatant, adding sulfur, and ageing for more than 3 months to obtain the mulberry fruit wine.

In the invention, the mulberry varieties are as follows: more than ten kinds of Guangdong mulberry, Deguo No. 1, Jialing 30, Jialing 40, Hongguo No. 2, Taiwan four seasons mulberry, white jade king or pearl white. Preferably, the variety of the mulberry is Guangdong mulberry.

In some embodiments, the fragmenting comprises: crushing mulberry once, separating juice and residue, crushing the crushed residue twice to obtain mulberry juice and mulberry residue, and mixing the mulberry juice and mulberry residue obtained by crushing twice.

In the present invention, the marc is not separated after crushing, and then the cap is removed the next day of the main fermentation. In this way, the bioactive components in the fruit residues and the residual fruit juice in the fruit residues can be fully dissolved; however, researches show that if only fruit juice is used for fermentation, the fruit residues are generally discarded, so that resource waste and great environmental pollution are caused; if the fruit residues are fermented into the fertilizer, local fermentation is needed, the fermentation effect is different, the waste of land resources is caused, and the stacking and fermentation of the fruit residues still have certain influence on the environment; the whole fruit is fermented, the pomace is fermented together, the time and the cost of intermediate fermentation of fertilizer producers are saved, meanwhile, mulberry fermentation liquor obtained by further centrifuging the pomace obtained by fermentation can be processed into mulberry brandy, and the centrifuged pomace is fermented uniformly, so that the fermentation effect is consistent, the mulberry brandy can be directly blended into fertilizer, the operation is easier, the mulberry resource is utilized to the maximum extent, and the green cyclic utilization of the mulberry resource is formed.

In some embodiments, in the enzymolysis step, the amount of sulfur added into the fermentation broth is 60mg/kg, leaching is performed for 12 hours at 4-6 ℃ after sulfur is added, and then heating to 20 ℃ is performed for enzymolysis for 4 hours by adding pectinase.

In some embodiments, the addition of pectinase comprises: dissolving pectinase in 10 times of purified water, and directly adding into Mori fructus fermentation broth.

In some embodiments, the yeast is a fruit wine yeast

D254^TMThe addition amount is 0.2 g/kg; the fermentation auxiliary agent is

Vitapermanent, added in an amount of 0.2 g/kg.

In some embodiments, the yeast is activated prior to addition; the activation of the yeast comprises: adding yeast into 5% of sucrose water 10 times of the weight of the yeast, activating for 10-20 min at 35-36 ℃, cooling to 25 ℃, standing for 5min, then adding the yeast to the surface of the fermentation liquor, standing for 5min, and mixing with the fermentation liquor.

In some embodiments, the post-fermentation is completed, and the supernatant is filled into an aging tank to enter an aging stage, and the aging stage is inverted once every 3 months to remove wine lees; after the aging, the wine body is cooled to 2-4 ℃, kept stand and clarified for 1d, and then filtered by a 0.22 mu m membrane.

In some embodiments, the alcoholic cap removed in the main fermentation step is distilled and aged to obtain morous brandy;

the temperature of the distilled cooling water is 25-35 ℃, and the distillation comprises the following steps:

cutting off the wine head: removing wine with the alcoholic strength of more than 55% vol;

middle-stage liquor: collecting wine with the alcoholic strength of 40-55 vol%;

cutting the tail of wine: and collecting wine with the alcoholic strength of 20-40 vol% and then re-steaming.

Before the distillation, the method comprises the step of centrifuging the wine cap to obtain wine liquid.

The post-fermentation and ageing stages are all in a low-temperature environment, the clarification method adopts a physical clarification method combining static clarification, low-temperature clarification and membrane filtration, the losses of components such as anthocyanin and pigment of the fruit wine are small, the light transmittance is high, and the fruit wine is clear and bright. In some embodiments, the aging comprises aging wine with the alcoholic strength of 40-55 vol% for 5 months at 15-20 ℃ in the presence of oak chips, removing the oak chips, aging for 1-2 years at 15-20 ℃ in a sealed manner, and filtering with a 0.22-micron membrane to obtain the mulberry brandy. The oak is a heavy oak chip.

The mulberry fruit wine or mulberry brandy prepared by the method is provided.

The mulberry fruit wine or mulberry brandy prepared by the method has higher quality and good sense. Especially, the mulberry fruit wine has advantages in sense and high yield. Wherein, the yield of the mulberry fruit wine is 50 percent to 55 percent, and the yield of the brandy is 1 ton of pomace, so that 50L of mulberry brandy can be obtained.

This is inseparable from the appropriate brewing conditions. The mulberry fruit wine preparation method provided by the invention selects varieties with high anthocyanin content, high sugar content and high juice yield, and selects mulberry varieties with good fruit wine flavor and quality in fermentation experiments of different varieties of mulberries. In the brewing process, secondary squeezing is adopted, the juice yield of the mulberry is improved, and after sulfur dioxide and pectinase are added to mulberry fermentation liquor obtained by fermenting the whole mulberry and extracting the mulberry at low temperature, substances such as juice, pigment, anthocyanin and the like in the mulberry are further dissolved out, so that the anthocyanin and dry extract content in the fruit wine is higher than that obtained by fermenting the whole mulberry. The anthocyanin in the mulberry fermentation liquid is better reserved by adopting temperature-controlled fermentation. The removal time and the method of a large amount of pomace are optimized, and the situation that anthocyanin and dry extract are not fully dissolved out due to too short retention time of the pomace is ensured; and the fruit wine has poor flavor due to long retention time. The removed pomace is distilled and processed into mulberry brandy, so that the yield of fruit wine is further improved, and the economic benefit is improved.

Drawings

FIG. 1 shows the detection report of the whole juice fermented (temperature controlled) fruit wine, wherein 1-a-1-c sequentially show 1-3 pages of the detection report;

FIG. 2 shows a detection report for whole fruit fermented (temperature controlled) fruit wine, wherein 2-a-2-c sequentially show 1-3 pages of the detection report;

FIG. 3 shows the test report for the whole fruit fermented (temperature-uncontrolled) fruit wine, wherein 3-a to 3-c show 1 to 3 pages of the test report in sequence;

Detailed Description

The present invention provides nucleic acid sequences and their use as promoters, which can be achieved by one skilled in the art with the appropriate modification of process parameters in view of the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.

The test materials adopted by the invention are all common commercial products and can be purchased in the market.

Wherein the sulfur dioxide effervescent tablet is prepared by adding

5EFFERVESCENT release sulfur dioxide; the pectinase is pectinase

Enzymic; the active dry yeast for brewing wine is fruit wine yeast

D254^TM(ii) a The fermentation auxiliary agent is

Vitaverment, which comprises vitamin B1 and ammonium salt; the oak chips are French heavy oak chips FGJK-CFFR +.

The invention is further illustrated by the following examples:

example 1 study of processing quality of different varieties of Morus alba

The juice yield, the pH value and the sugar acid content of different varieties of mulberries are detected, and the results are shown in table 1:

TABLE 1 quality index of different varieties of Mori fructus

Note: in the table, multiple comparisons are performed by a Duncan method, wherein the shoulder marks of the same row with different capital letters indicate that the difference between groups is significant (p is less than 0.01), the lower-case letters corresponding to the capital letters indicate that the difference between groups is significant (p is less than 0.05), and the same capital letters indicate that the difference between groups is not significant (p is more than 0.05).

Wherein, the highest and lowest juice yield are respectively dozens of Guangdong mulberry (75.25%) and No. 1 Deguo (54.55%).

Fruit wine, fruit juice and other products require high sugar content, and the total sugar content plays an important role in influencing the flavor of the fruit. The varieties with the total sugar and reducing sugar content of more than 10g/100g include Dadeca Yuehei, Deguo No. 1 and pearl white.

The specific gravity is an important parameter for fermenting the fruit wine, and the processing of the fruit wine is influenced by the specific gravity. The specific gravity of 8 varieties except the red fruit No. 2 is close to 1.000kg/L, and the specific gravity of the other varieties is more than 1.000 kg/L.

TABLE 2 comparison of procyanidin content in different varieties of Mori fructus

Index (I)

Ten big Guangdong mulberry

De fruit No. 1

Jialing 30

Jialing 40

Haw No. 2

Taiwan mulberry leaf

White jade king

Pearl white

Procyanidin content g/100g

0.674

0.627

0.474

0.361

0.335

0.384

＜0.015

Procyanidin is natural antioxidant of plant polyphenols, can be heated under acidic condition to generate anthocyanidin, and has effects of scavenging free radicals and inhibiting lipid peroxidation. As can be seen from Table 2, the mulberry has abundant procyanidine content, the content is 0-0.68 g/100g, the content difference of procyanidine of different varieties is large, and the varieties ranked in the first three are Guangdong mulberry, Deguo No. 1 and Jialing 30.

The cluster analysis of the processing characteristics among different mulberry varieties results in that: the three types have 1 variety, the comprehensive score of Guangdong mulberry and Guangdong mulberry is the highest (0.75), the processing is most suitable, the main characteristics are high juice yield, high procyanidine content, high soluble total sugar and reducing sugar, and the titratable acid content is 8.33 g.L^-1The mulberry is suitable for being processed into fruit juice and fruit wine.

Example 2

Optimizing a fermentation process by taking Dadeca yunnanensis as a target, wherein the optimization process mainly comprises the following steps of treating pomace and fruit juice in a main fermentation stage:

1. selecting mulberry wine raw materials: selecting fruits with reducing sugar content of more than 8g/100g and no decay and deterioration;

2. crushing: crushing by a crusher, separating juice and residue, and crushing the primary crushed residue in the crusher for the second time to obtain mulberry juice and mulberry residue respectively;

3、

3.1 fermenting the whole fruit: mixing Mori fructus juice and Mori fructus residue in fermentation tank to obtain Mori fructus fermentation broth;

3.2 fermenting the whole juice: the mulberry juice obtained by two-time crushing is transferred into a fermentation tank to be used as fermentation liquor.

4. Component blending: adding white granulated sugar with theoretical alcoholic strength reaching 12% vol, and uniformly stirring by an intelligent stirring system;

5. the low-temperature leaching and the addition of the sulfur dioxide effervescent tablets are carried out simultaneously: through an intelligent water circulation system, mulberry fermentation liquor is subjected to static low-temperature leaching for 12 hours at the temperature of 4-6 ℃, sulfur dioxide effervescent tablets with the dosage of 60mg/kg are added at the beginning of leaching, and the mulberry fermentation liquor is subjected to sealing treatment, and has the effects of sterilization, clarification, oxidation resistance, dissolution of pigments and tannin substances and the like.

Sulfur dioxide effervescent tablets: directly placing the sulfur dioxide effervescent tablet into the mulberry fermentation liquor prepared by the components, immediately sealing, and uniformly stirring;

6. and (3) treating pectinase: after the leaching is finished, the fermentation liquor is heated to 20 ℃, and pectinase is added for enzymolysis for 4 hours, wherein the dosage is 20 mg/kg.

And (3) pectinase: dissolving pectinase in 10 times of purified water, directly adding into Mori fructus fermentation liquid, stirring, improving Mori fructus soaking effect, squeezing, and rapidly clarifying Mori fructus juice;

7. temperature-controlled fermentation is accompanied by yeast and fermentation auxiliary agent addition, and the whole process of the main fermentation stage is as follows: after the temperature of the mulberry fermentation liquor is raised to 20-25 ℃, adding activated brewing active dry yeast and fermentation auxiliary agent, wherein the dosage is 0.2g/kg, sealing, and uniformly stirring;

activation of active dry yeast for brewing: putting active dry yeast into 5% sugar water which is 10 times of the weight of the active dry yeast, activating at 35-36 ℃, fully stirring and dissolving until a layer of creamy substance appears on the surface of the yeast liquid, successfully activating for about 15min, adjusting the temperature of the activated yeast liquid to 25 ℃ by using circulating water, standing for 5min, adding the activated yeast liquid to the surface of mulberry fermentation liquid, standing for 5min, and uniformly stirring.

Fermentation aid: and (3) fully dissolving the fermentation auxiliary agent by using part of mulberry fermentation liquor, adding the dissolved fermentation auxiliary agent into the mulberry fermentation liquor, and fully and uniformly stirring.

8. A main fermentation stage:

8.1 controlling the temperature to ferment, controlling the temperature to 20-25 ℃ in the whole process, pressing a wine cap every 12 hours 2 days before fermentation, and then intelligently stirring for 30 min; removing pomace from the upper part of the fermentation tank when the mulberry is fermented for the next day, supplementing white granulated sugar, continuing sealed fermentation until the total sugar content in the mulberry fermentation liquor is below 1% and the alcoholic strength is about 12% vol, ending the main fermentation, taking supernate to obtain a fermentation product, controlling the fermentation temperature in the whole main fermentation process to be 20-25 ℃, and jointly completing temperature-controlled fermentation through a temperature and humidity detection control system and an intelligent circulating water system.

8.2 fermenting without controlling the temperature, pressing a wine cap every 12h 2 days before fermenting, and intelligently stirring for 30 min; removing fruit residues through the upper part of the fermentation tank when the fermentation is carried out for the next day, supplementing white granulated sugar, continuing sealed fermentation until the total sugar content of the mulberry fermentation liquor is below 1% and the alcoholic strength is about 12% vol, ending the main fermentation, and taking supernatant to obtain a fermentation product.

9. And (3) post-fermentation stage: transferring the fermentation product into a ceramic tank of a wine cellar at 15-20 ℃ through a transfer pump, filling and sealing the ceramic tank, and performing after-fermentation; standing for one month, collecting supernatant, filling in aging tank, adding sulfur dioxide effervescent tablet with dosage of 30mg/kg, fermenting, sealing, and aging;

10. and (3) ageing stage: standing for clarification, and pouring out every 3 months and 6 months to remove wine lees to obtain clarified wine;

11. and (3) post-treatment: blending the clarified wine, cooling to 2-4 ℃, standing for clarification for 24 hours, and filtering and sterilizing by a 0.22 mu m membrane filter to obtain the fermented mulberry fruit wine with the yield of 50-55%.

TABLE 3 product testing data

Index (I)	Full juice fermentation (temperature control)	Whole fruit fermentation (temperature control)	Whole fruit fermentation (temperature-controllable)
				The total amount of anthocyanidin is mg/kg	133.69	352.78	178.73
Total acid g/L	5.1	8.5	8.1
				g/L of volatile acid	0.6	0.6	0.8
g/L of dry extract	11.6	21.9	20.8

The obtained mulberry wine finished product: the alcohol content (vol) is more than or equal to 11.5 and less than or equal to 12.4, the total acid (g/L, counted by tartaric acid) is more than or equal to 8 and less than or equal to 9, the volatile acid (g/L) is more than or equal to 1, the dry extract (g/L) is more than or equal to 20, salmonella and staphylococcus aureus (CFU/mL) are not detected, and the total anthocyanin amount (mg/kg) is more than or equal to 300.

Wherein, the detection result of the full juice fermented (temperature controlled) fruit wine is shown in figure 1;

the detection result of the whole fruit fermented (temperature controlled) fruit wine is shown in figure 2;

the detection results of the whole fruit fermented (temperature-controlled) fruit wine are shown in figure 3.

According to the results shown in table 3 and fig. 1-3, the fruit wine prepared by whole fruit fermentation (temperature control) has the best quality, and the fruit wine is used as the optimal scheme of fermentation for further research.

Example 3

Adopting the preferred range in the embodiment 2, different mulberry varieties are brewed with fruit wine, and the method comprises the following steps:

3. fermenting the whole fruit: mixing Mori fructus juice and Mori fructus residue in fermentation tank to obtain Mori fructus fermentation broth;

8. A main fermentation stage: pressing a wine cap every 12h 2 days before fermentation, and intelligently stirring for 30 min; removing pomace from the upper part of the fermentation tank when the mulberry is fermented for the next day, supplementing white granulated sugar, continuing sealed fermentation until the total sugar content of the mulberry fermentation liquor is below 1% and the alcoholic strength is about 12% vol, ending the main fermentation, taking supernate to obtain a fermentation product, controlling the fermentation temperature in the whole main fermentation process to be 20-25 ℃, and jointly completing temperature-controlled fermentation through a temperature and humidity detection control system and an intelligent circulating water system.

TABLE 4 fermentation experimental data of different varieties of mulberry and mulberry wine

Index (I)	Fermentation stage	Ten big Guangdong mulberry	De fruit No. 1	Jialing 30	Jialing 40	Haw No. 2	Taiwan mulberry leaf	White jade king	White pearl
										pH	End of main fermentation	3.90	4.23	3.95	3.87	3.46	3.50	3.63	3.66
Titratable acid g/L	End of main fermentation	7.48	6.11	17.14	8.87	13.15	12.66	8.94	9.75
										Alcohol content%	End of main fermentation	10.90	10.55	7.50	11.15	9.55	11.65	10.25	10.60
g/L of volatile acid	End of main fermentation	1.39	0.85	8.15	2.13	1.12	0.63	0.69	1.60

According to the data, the titratable acid, the alcoholic strength and the volatile acid of the mulberry wine fermented by the Dadeca yuen are in ideal ranges, and the titratable acid of the fruit wine is in an ideal range of 7-9 g/L; the wine fermented by the jialing 30 has too low alcoholic strength (white granulated sugar is added according to 12% vol alcoholic strength), high titratable acidity and high volatile acidity, and is not suitable for drinking.

Therefore, by combining the test data, pilot test in each stage, cost of finished mulberry and detection report in examples 2-3, the Dadeca Yuexian was selected as the mulberry variety for the fruit wine fermentation, and the brewing scheme was selected for full juice fermentation (temperature control).

The foregoing is only a preferred embodiment of the present invention, and it should be noted that it is obvious to those skilled in the art that various modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.

Claims

1. A preparation method of mulberry fruit wine is characterized by comprising the following steps:

and (3) after-fermentation: and sealing and standing the supernatant at 15-20 ℃ for 1 month to obtain the supernatant, adding sulfur into the supernatant, and ageing the supernatant for more than 3 months to obtain the mulberry fruit wine.

2. The method for preparing mulberry fruit wine according to claim 1, wherein the varieties of mulberry fruit are: more than ten kinds of Guangdong mulberry, Deguo No. 1, Jialing 30, Jialing 40, Hongguo No. 2, Taiwan four seasons mulberry, white jade king or pearl white.

3. The method for preparing morous fruit wine according to claim 1, wherein the crushing comprises: crushing mulberry once, separating juice and residue, crushing the crushed residue twice to obtain mulberry juice and mulberry residue, and mixing the mulberry juice and mulberry residue obtained by crushing twice.

4. The preparation method of mulberry fruit wine according to claim 1, wherein in the step of enzymolysis, the amount of added sulfur in the fermentation broth is 60mg/kg, the fermentation broth is extracted at 4-6 ℃ for 12h after the sulfur is added, and then the temperature is raised to 20 ℃ and pectinase is added for enzymolysis for 4 h.

5. The method for preparing morous fruit wine according to claim 1, wherein the yeast is fruit wine yeast

D254^TMThe addition amount is 0.2 g/kg; the fermentation auxiliary agent is

Vitapermanent, added in an amount of 0.2 g/kg.

6. The method for preparing morous fruit wine according to claim 1 or 5, wherein the yeast is activated before being added; the activation of the yeast comprises: adding yeast into 5% of sucrose water 10 times of the weight of the yeast, activating for 10-20 min at 35-36 ℃, cooling to 25 ℃, standing for 5min, then adding the yeast to the surface of the fermentation liquor, standing for 5min, and mixing with the fermentation liquor.

7. The method for preparing morous fruit wine according to claim 6, wherein the supernatant after the post-fermentation is completed is filled in an aging tank and enters an aging stage, and the aging stage is inverted every 3 months to remove wine lees; after the aging, the wine body is cooled to 2-4 ℃, kept stand and clarified for 1d, and then filtered by a 0.22 mu m membrane.

8. The process for preparing morous alba wine according to claim 1, wherein the caps removed in the main fermentation step are distilled and aged to obtain morous alba;

middle-stage liquor: collecting wine with the alcoholic strength of 40-55 vol%;

9. The method for preparing mulberry fruit wine according to claim 8, wherein the aging comprises aging a wine liquid with an alcohol content of 40-55 vol% at 15-20 ℃ for 5 months in the presence of oak chips, removing the oak chips, aging under sealed conditions at 15-20 ℃ for 1-2 years, and filtering with a 0.22 μm membrane to obtain mulberry brandy.

10. Mulberry fruit wine or Mulberry brandy obtained by the method of any one of claims 1 to 9.