Plant-soaked solution comprising tagatose, and method for producing same

Abstract

The present application relates to a plant-soaked solution comprising sugars containing tagatose and to a method for producing same.

Claims

1. A plant-soaked solution composition comprising a plant and a saccharide containing tagatose, wherein the plant-soaked solution is obtained by: mixing a plant with saccharide containing tagatose; and soaking the plant with the saccharide containing tagatose for 15 days to 6 months to provide the plant-soaked solution composition, wherein the ratio of the saccharide containing tagatose:plant soaked solution is 33 to 67 parts by weight:100 parts by weight of the soaked solution, wherein the tagatose is present in an amount of 1 to 100 parts by weight:100 parts by weight of the saccharide containing tagatose, based on dry solids, and wherein the plant comprises fruit.

2. The plant-soaked solution composition of claim 1, wherein the saccharide does not contain sucrose.

3. The plant-soaked solution composition of claim 1, wherein the fruit comprises lemon.

4. The plant-soaked solution composition of claim 1, which is a health functional food for blood glucose regulation.

5. The plant-soaked solution composition of claim 3, which is a health functional food for antioxidation.

6. The plant-soaked solution composition of claim 1, wherein the fruit comprises plum.

7. The plant-soaked solution composition of claim 1, wherein the fruit comprises one or more fruits selected from the group consisting of Prunus mume, citrus, lemon, citron, grapefruit, lime, quince, schizandra, rubus coreanus, pear, apple, grape, mulberry, blueberry, mango, peach, Prunus salicina, apricot, sweet persimmon, banana, and jujube.

8. A method of preparing a plant-soaked solution, comprising Preparing the plant-soaked solution composition of claim 1, filtering the plant-soaked solution composition, and collecting the filtered solution, wherein the filtered solution is the plant-soaked solution.

9. The method of claim 8, wherein the plant-soaked solution further comprises polyphenol and the content of total polyphenol in the plant-soaked solution is 0.6 mg/ml or more.

10. The method of claim 8, wherein the plant-soaked solution contains at least 0.67 mg/ml polyphenol.

11. A method of extracting polyphenol from fruit in the plant-soaked solution composition of claim 1, comprising adding saccharide containing tagatose to the plant.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is photographs of tissues of Prunus mumes soaked; FIG. 1A shows the tissue of Prunus mume steeped in sucrose for 3 months; and FIG. 1B shows the tissue of Prunus mume steeped in tagatose for 3 months.

(2) FIG. 2 is a graph showing changes in the contents of tagatose and free saccharides (sucrose, glucose, and fructose) in the soaked solutions prepared using sucrose and tagatose, respectively.

(3) FIG. 3 is a diagram illustrating an example of a randomized cross-over design study.

(4) FIG. 4 is a graph comparing the blood glucose value results of Comparative example 3 with those of Experimental example 3.

(5) FIG. 5 is a graph comparing the AUC result of Comparative example 3 with that of Experimental example 3.

MODE FOR CARRYING OUT THE INVENTION

(6) The present invention will be described in detail with reference to the following Examples, but the following Examples are provided by way of illustration, and the present invention is not limited thereto.

Preparation Example 1: Preparation of Prunus mume-Soaked Solution Using Tagatose

(7) Domestic Prunus mume (green mumes) were washed cleanly with distilled water and allowed to dry naturally for one day. Then, stems were removed and 0.95 kg was added to a soaking vessel (glass material, capacity 5 kg) which was heated in boiling water at 100 C. for 10 minutes and naturally dried. Thereafter, 0.95 kg of tagatose (crystalline form, purity of 99% or more, CJ CheilJedang) was added, and the same amount of Prunus mumes and the same amount of tagatose were added in serial order one more time. Thereafter, the resulting products were maintained for 1 month, 2 months, and 3 months in a cold dark place condition (4 C., light-blocked condition, Korean food code), and then filtered with a sieve (8 mesh). The remaining original liquid was collected to prepare Prunus mume-soaked solutions [Experimental example 1 (maintenance for 1 month), Experimental example 2 (maintenance for 2 months) and Experimental example 3 (maintenance for 3 months)].

Preparation Example 2: Preparation of Lemon-Soaked Solution Using Tagatose

(8) Lemon was washed with baking soda, and then, washed with distilled water to remove residual pesticide on the surface of lemon, and dried naturally for one day, and then sliced to a thickness of 0.5 to 2 cm, and seed was removed. 400 g of the resulting lemon was added to a a soaking vessel (glass material, capacity 1 kg) in the same manner as in Preparation example 1, and then 400 g of tagatose was added, and the same amount of lemon and the same amount of tagatose were added in serial order one more time. Thereafter, the resulting product was maintained for 15 days under cold dark condition (4 C., light-blocked condition, Korean food code), and then filtered with a sieve (8 mesh). The remaining original liquid was collected to prepare lemon-soaked solution (Experimental example 4).

Preparation Example 3: Preparation of Prunus mume-Soaked Solution and Lemon-Soaked Solution Using Sucrose

(9) Prunus mume-soaked solutions [Comparative example 1 (maintenance for 1 month), Comparative example 2 (maintenance for 2 months), and Comparative example 3 (maintenance for 3 months)] and lemon-soaked solution were prepared in the same manner as in Preparation examples 1 and 2 by substituting sucrose (crystalline-type white sucrose, CJ CheilJedang) for tagatose.

Example 1: Evaluation of Prunus mume-Soaked Solution

(10) 1-1. Evaluation of Tissue Change

(11) In the production of the Prunus mume-soaked solution, the moisture in the original material of Prunus mume is drained from the inside to outside of the tissue by the osmosis principle so that the tissue contracts. Therefore, it is usually judged that the more tissue of soaked Prunus mume contracts, the further the soaking progresses. Thus, changes in Prunus mume tissues after 3 months from the start of soaking in Preparation example 1 and Preparation example 3 were visually observed.

(12) As a result, it was found that the tissue of the soaked Prunus mume of Preparation example 1 (FIG. 1B) contracted at the same level as the level of the soaked Prunus mume of Preparation example 3 (FIG. 1A) (FIG. 1).

(13) 1-2. Evaluation of Physical Properties (Solid Content, pH, and T-Color) of Soaked Solution

(14) In order to verify whether the soaking is carried out at the same level as in the Prunus mume-soaked solution using sucrose when tagatose is used for preparation of Prunus mume-soaked solution, physical properties (solid content, pH, and T-color) of Comparative examples 1, 2, and 3, and Experimental examples 1, 2, and 3 were measured and compared.

(15) Specifically, the solid content (g content of solids dissolved in 100 g of soaked solution) was measured using a refractometer (ATAGO, Automatic Digital Refractometer RX-5000a); the pH was measured with a digital pH meter (METTLER TOLEDO, SEVEN COMPACT with InLab Viscous Pro pH); and 1 cm cells were filled with Experimental examples 1, 2, and 3, and Comparative examples 1, respectively, and then the T-color (transmission color) was measured at a wavelength of 420 nm using a spectrophotometer (HITACHI, Double Beam Spectrophotometer U-2900) calibrated with distilled water. Statistical analysis was performed using SAS 9.1 program (SAS Inc., Cary, NC, USA) and the results were analyzed using the t-test method. All analyses were tested at the significance level of p<0.05.

(16) As a result, there was no significant difference in Brix %, pH, and T-color, respectively, between Experimental examples 1, 2 and 3, and Comparative examples 1, 2 and 3 (Table 1). As for the solid content, like the conventional soaked solution, 50 Brix % or more was exhibited. The pH and T-color were not statistically significantly different between Experimental examples and Comparative examples (on the basis of 95% significance probability, p>0.05).

(17) As a result, it could be found out that when the Prunus mume-soaked solution was prepared using tagatose, the soaked solution of the same level, in terms of physical properties, as the Prunus mume-soaked solution using sucrose, was embodied.

(18) TABLE-US-00001 TABLE 1 Solid content (Brix %) pH T-color (%) Comparative Comparative Comparative Comparative Comparative Comparative Comparative Comparative Comparative example 1 example 2 example 3 example 1 example 2 example 3 example 1 example 2 example 3 57.20 57.55 56.50 2.62 2.54 2.65 83.15 80.10 49.90 Experimental Experimental Experimental Experimental Experimental Experimental Experimental Experimental Experimental example 1 example 2 example 3 example 1 example 2 example 3 example 1 example 2 example 3 53.13 53.23 52.90 2.64 2.66 2.71 79.17 68.40 44.50
1-3. Determination of Citric Acid Content in the Soaked Solution

(19) In order to verify whether the soaking is carried out at the same level as in the Prunus mume-soaked solution using sucrose and the useful components are extracted when tagatose is used for preparation of Prunus mume-soaked solution, the content of citric acid, which is one of main useful components of Prunus mume, in the Prunus mume-soaked solution was measured by high performance liquid chromatography (HPLC).

(20) The test solution was prepared by placing 1 g of sample of Experimental examples 1, 2, and 3, and Comparative examples 1, 2, and 3, in a 50 mL constant volume flask, dissolving it in distilled water to adjust the total volume of the solution to 50 mL (20 g/L), and then filtering it through a 0.2 m filter. The standard solution was prepared by placing 0.5 g of citric acid standard material (citric acid monohydrate, Sigma Aldrich) in a 50 mL constant volume flask, dissolving it in distilled water to adjust the total volume of the solution to 50 mL (10 g/L), diluting it to concentrations of about 0.3125 g/L, 0.625 g/L, 1.25 g/L, 2.5 g/L, 5 g/L and 10 g/L, and filtering it through a 0.2 m filter.

(21) The prepared test solutions and standard solutions were analyzed using HPLC (Alliance, Waters, e2695 Separation Modules, USA/Waters column Heater Module/RI detector Water 2414/Empower Software) under the conditions shown in Table 2 below.

(22) TABLE-US-00002 TABLE 2 Mobile phase 5 mM H.sub.2SO.sub.4 Column 300 mm 7.8 mm Amninex 87H (Bio Rad) Flow rate 0.6 mL/min Temperature 35 C. Injection volume 20 L Detector DAD (Diode Array Detector)

(23) After HPLC analysis, a calibration curve was prepared with the citric acid content (g/L) as the abscissa axis and the area of the chromatogram as the ordinate axis. The citric acid areas Experimental examples 1, 2 and 3 and Comparative examples 1, 2 and 3 were read, and the citric acid contents were calculated from the calibration curve. As a result, it was found out that there was no significant difference in the contents of citric acid in Experimental examples 1, 2 and 3 and Comparative examples 1, 2 and 3, respectively (Table 3). As a result, it could be found out that when the solution was prepared using tagatose, the soaked solution of the same level, in terms of useful components, as the soaked solution using sucrose was embodied.

(24) TABLE-US-00003 TABLE 3 Citric acid content (g/L) Comparative Comparative Comparative example 1 example 2 example 3 16.73 21.69 17.55 Experimental Experimental Experimental example 1 example 2 example 3 18.25 19.82 16.08
1-4. Organoleptic Evaluation

(25) Samples of Experimental example 3 and Comparative example 3 were diluted in lukewarm water at a ratio of 4:6, and each of the diluted samples was subjected to organoleptic evaluation by a trained panel of 17 evaluators for five preference attributes (flavor/color/sour taste/sweet taste/overall preference).

(26) [Definition of Organoleptic Evaluation Terms]

(27) 1) Flavor preference: Personal preference for intrinsic flavor of soaked solution 2) Sweet taste preference: Personal preference for sweet taste 3) Sour taste preference: Personal preference for sour taste 4) Color preference: Personal preference for lightness and darkness of color of soaked solution 5) Overall preference: Overall personal preference

(28) As a result, it was found that color preference for Experimental example 3 was significantly higher than that for Comparative example 3, and other preference attributes except for color preference were similar (Table 4).

(29) TABLE-US-00004 TABLE 4 Comparative Experimental 5 point scale example 3 example 3 p-value Flavor 2.6 2.5 0.234 preference Sweet taste 2.6 2.5 0.891 preference Sour taste 2.6 2.3 0.396 preference Color 1.9 2.8 0.001* preference Overall 2.4 2.2 0.425 preference *p < 0.05
1-5. Determination of Tagatose Decomposition

(30) In order to verify whether the calorie reduction effect of tagatose and the inherent functionality were maintained in the soaked solution prepared using tagatose, it was determined whether tagatose used in the preparation decomposed or not in the soaked solution.

(31) Specifically, the contents of tagatose and free saccharides (sucrose, glucose, and fructose) in Experimental examples 1, 2, and 3, and Comparative examples 1, 2, and 3 were measured by using high performance liquid chromatography (HPLC). The test solution was prepared by placing 1 g of sample of Experimental examples 1, 2, and 3, and Comparative examples 1, 2, and 3, in a 50 mL constant volume flask, dissolving it in distilled water to adjust the total volume of the solution to 50 mL (20 g/L), and then filtering it through a 0.2 m filter. The standard solutions were prepared by placing 1 g of each standard material, sucrose (SigmaS7903, CAS No. 57-50-1), glucose (SigmaG7528, CAS No. 50-99-7), fructose (SigmaF0127, CAS No. 57-48-7) and tagatose (Sigma75935, CAS No. 87-81-0) in a 50 mL constant volume flask, dissolving it in distilled water, diluting it to concentrations of about 0.625 g/L, 1.25 g/L, 2.5 g/L, 5 g/L, 10 g/L and 20 g/L, and filtering it through a 0.2 m filter.

(32) The prepared test solutions and standard solutions were analyzed using HPLC (Alliance, Waters, e2695 Separation Modules, USA/Waters column Heater Module/RI detector Water 2414/Empower Software) under the conditions shown in Table 5 below.

(33) TABLE-US-00005 TABLE 5 Mobile phase Distilled water (HPLC Grade) Column 7.8 mm 300 mm Amninex HPX87C (Bio Rad) Flow rate 0.6 mL/min Temperature 80 C. Injection volume 20 L Detector Differential refractometer (RID: Refractive Index Detector)

(34) After HPLC analysis, a calibration curve was prepared with the contents of tagatose and free saccharides (sucrose, glucose, and fructose) (g/L) as the abscissa axis and the area of the chromatogram as the ordinate axis. The areas of tagatose and free saccharides of test solutions of Experimental examples 1, 2 and 3, and Comparative examples 1, 2 and 3 were read, and the contents of tagatose and free saccharides were obtained from the calibration curve (error range: 5%).

(35) As a result, it could be found that in Comparative example, sucrose decomposed into glucose and fructose, and in particular, in Comparative example 3, about 89.6% of the initial sucrose content at the soaking decomposed into glucose and fructose, but in all Experimental examples 1, 2, and 3, tagatose did not decompose and remained intactly (Table 6 and FIG. 2). Therefore, it could be found that in the conventional soaked solution prepared using sucrose, sucrose decomposed to glucose and fructose during the soaking period, which, when consumed, caused a rapid rise in blood glucose and provided high calorie (4 Kcal/g), but the soaked solution prepared using tagatose did not exhibit change in components during the soaking, and thus, could maintain the intrinsic calorie reduction effect (1.5 Kcal/g) and functionality of tagatose.

(36) TABLE-US-00006 TABLE 6 Sucrose content Glucose content Fructose content Tagatose content Compar- Compar- Compar- Compar- Compar- Compar- Compar- Compar- Compar- Compar- Compar- Compar- ative ative ative ative ative ative ative ative ative ative ative ative example 1 example 2 example 3 example 1 example 2 example 3 example 1 example 2 example 3 example 1 example 2 example 3 30.8 8.4 3.2 15.4 33.0 38.0 13.0 22.1 24.0 Experi- Experi- Experi- Experi- Experi- Experi- Experi- Experi- Experi- Experi- Experi- Experi- mental mental mental mental mental mental mental mental mental mental mental mental example 1 example 2 example 3 example 1 example 2 example 3 example 1 example 2 example 3 example 1 example 2 example 3 55.3 56.7 57.7 (Unit: g/100 g of soaked solution)
1-7. Determination of Total Polyphenol Content in the Soaked Solution

(37) In order to verify the functionality of Prunus mume-soaked solution, the content of polyphenol in the soaked solution was measured using Microplate Reader (Powerwave XS, BioTek, USA).

(38) 2% Sodium carbonate reagent is prepared by placing 2 g of sodium carbonate (Sigma 223484, CAS No. 497-19-8) in a 100 mL constant volume flask and adding distilled water thereto to adjust the total volume to 100 mL Folin-Ciocalteu's phenol reagent (Sigma F9252-1L) and distilled water were mixed at a ratio of 1:1 to prepare 50% Folin-Ciocalteu's phenol reagent. It was wrapped with aluminum foil to prevent light from being transmitted.

(39) As for the test solutions, each of Experimental examples 1 to 3 and Comparative examples 1 to 3 was diluted with distilled water at a ratio of 1:1, and 0.1 mL of each solution was mixed with 0.1 mL of 50% Folin-Ciocalteu's phenol reagent and 2 mL of 2% sodium carbonate. Then, the test solutions were allowed to stand in a dark place for 30 minutes and absorbance was measured at 750 nm.

(40) The standard solution was prepared by taking 0.4 g of gallic acid (Sigma G7384, CAS No. 149-91-7) in a 100 mL contant volume flask, adding distilled water thereto to adjust the total volume of the solution to 100 mL, and diluting the solution to concentrations of 31.25 ppm, 62.5 ppm, 125 ppm, 250 ppm and 500. The standard solution was mixed with the reagents in the same manner as the test solutions, and allowed to stand in a dark place for 30 minutes and then, absorbance was measured at 750 nm.

(41) After the absorbance measurement, a calibration curve was prepared with the absorbance of the standard solution as the abscissa axis and the concentration of the standard solution as the ordinate axis. The total polyphenol content of each of Experimental examples 1 to 3 and Comparative examples 1 to 3 was calculated by using Equation 6 below.
Total polyphenol content (mg/mL)=(ABC)/D[Equation 1] *A: Total amount of test solution (mL), B: Dilution factor, C: Total polyphenol concentration in the test solution (mg/mL), D: Collected sample amount (mL)

(42) As a result, it was found that the total polyphenol contents in Experimental examples 1 to 3 were significantly larger than those in Comparative Examples 1 to 3, which were stored for the same period of time (Table 7).

(43) Therefore, it could be found that in the preparation of Prunus mume-soaked solution, by using tagatose rather than sucrose, the soaked solution of higher content of polyphenol can be prepared.

(44) TABLE-US-00007 TABLE 7 Comparative Experimental Classification example 1 example 1 p value Total 0.45 0.67 0.023* polyphenol (mg/mL) Comparative Experimental example 2 example 2 p value 0.56 0.86 0.001* Comparative Experimental example 3 example 3 p value 0.52 0.73 0.000* *p < 0.05
1-7. Effect of Inhibiting Blood Glucose Rise

(45) In order to verify the health functionality of Experimental example 3, when Experimental example 3 and Comparative example 3 were consumed in a fasting state, changes in blood glucose were measured.

(46) Four normal subjects with a fasting glucose level of less than 126 mg/dL were selected to participate in a randomized cross-over design study (FIG. 3). In the morning on the day of the experiment, Experimental example 3 or Comparative example 3 diluted with water at a ratio of 1:4 was taken in a fasting state. Changes in blood glucose were measured using a glucose meter (OptiumXceed Blood Glucose Monitoring System, Abbott Diabetes CreInc., USA) at the time points (0 minute, 15 minutes, 30 minutes, 45 minutes, 60 minutes, and 90 minutes).

(47) In order to avoid a cross-over effect on the substance to be consumed, a wash-out period of one week was given, and Experimental example 3 or Comparative example 3 was taken in the same manner as above, and then, changes in blood glucose were measured and the area under the curve (AUC) was calculated by using the Trapezoidal Rule.

(48) As a result, it was found that Experimental example 3 significantly inhibited the rise in blood glucose after 15 minutes of intake by about 18% compared to Comparative example 3, and inhibited the rise in blood glucose after 30 minutes by about 17.8% (Table 8 and FIG. 4). The AUC of Experimental example 3 was lower than that of Comparative example 3 by about 10.9% (FIG. 5). Therefore, the intake of the soaked solution prepared using tagatose caused inhibition of the rapid rise in blood glucose, compared to the intake of the soaked solution prepared using sucrose, and thus, it was found that the intake of the soaked solution prepared using tagatose can help to inhibit the rapid rise in blood glucose after meals.

(49) TABLE-US-00008 TABLE 8 Classification 0 min 15 min 30 min 45 min 60 min 90 min Comparative 109.0 16.6 126.8 17.9 132.0 33.7 123.3 25.6 110 20.2 102.5 9.9 example 1 Experimental 109.5 9.5 103.8 13.0 108.5 15.1 106.3 8.0 99.8 7.3 103.8 8.3 example 1 p-value 0.932 0.008** 0.087* 0.163 0.229 0.391 *p < 0.1, **p < 0.05

Example 2: Evaluation of Lemon-Soaked Solution

(50) 2-1. Evaluation of Physical Properties (Solid Content, pH, and T-Color) of Soaked Solution

(51) In order to verify whether the soaking is carried out at the same level as in the lemon-soaked solution using sucrose when tagatose is used for preparation of lemon-soaked solution, physical properties (solid content, pH, and T-color) of Comparative example 4 and Experimental example 4 were measured and compared. The measurement methods of solid content, pH and T-color were the same as in Examples 1 to 2.

(52) As a result, as for the solid content, like the conventional soaked solution, 50 Brix % or more was exhibited. The pH and T-color were not statistically significantly different between Experimental example and Comparative example (on the basis of 95% significance probability, p>0.05) (Table 9).

(53) As a result, it could be found out that when the lemon-soaked solution was prepared using tagatose, the soaked solution of the same level, in terms of physical properties, as the lemon-soaked solution using sucrose, was embodied.

(54) TABLE-US-00009 TABLE 9 Solid content Classification (Brix %) pH T-color (%) Comparative 59.51 3.38 82.21 example 4 Experimental 56.46 3.42 86.38 example 4
2-2. Determination of Tagatose Decomposition

(55) In order to verify whether the low calorie of tagatose and the inherent functionality were maintained in the lemon-soaked solution prepared using tagatose, it was determined whether tagatose used in the preparation decomposed or not in the lemon-soaked solution.

(56) Specifically, the contents of tagatose and free saccharides (sucrose, glucose, and fructose) in Experimental example 4 and Comparative example 4 were measured by using HPLC. The measurement method and HPLC analysis conditions were the same as those of Example 1-5.

(57) As a result, it could be found that in Comparative example 4, sucrose decomposed to sucrose and fructose, but tagatose used in Experimental example 4 did not decompose and remained intactly (Table 10). Therefore, it could be found that as in Prunus mume-soaked solution, tagatose did not decompose also in lemon-soaked solution, and the lemon-soaked solution could maintain the intrinsic calorie reduction effect and functionality of tagatose.

(58) TABLE-US-00010 TABLE 10 Sucrose Glucose Fructose Tagatose Classification content content content content Comparative 49.4 3.6 3.1 example 4 Experimental 58.5 example 4 (Unit: g/100 g of soaked solution)
2-3. Determination of the Vitamin C Content in the Soaked Solution

(59) In order to verify whether the soaking is carried out at the same level as in the lemon-soaked solution using sucrose and the useful components are extracted when tagatose is used for preparation of lemon-soaked solution, the content of vitamin C, which is one of main useful components of lemon, in the lemon-soaked solution was measured by high performance liquid chromatography (HPLC).

(60) The test solution was prepared by placing 1 g of samples of Experimental example 4 and Comparative example 4, in a 50 mL constant volume flask, dissolving it in distilled water to adjust the total volume to 50 mL (20 g/L), and then filtering it through a 0.2 m filter. The standard solution was prepared by placing 0.01 g of vitamin C standard material (L-Ascorbic acid, SigmaA5960, CAS No. 50-81-7) in a 10 mL constant volume flask, dissolving it in distilled water to adjust the total volume of the solution to 10 mL (0.1 g/L), diluting it to 5 points of concentrations of about 0.00625 g/L, 0.0125 g/L, 0.025 g/L, 0.05 g/L, 0.1 g/L, and filtering it through a 0.2 m filter.

(61) The prepared test solutions and standard solutions were analyzed using HPLC (Alliance, Waters, e2695 Separation Modules, USA/Waters column Heater Module/RI detector Water 2414/Empower Software) under the analytic conditions shown in Table 11 below.

(62) TABLE-US-00011 TABLE 11 Mobile phase Acetonitrile 90% + formic acid 10% Column 250 mm 4.6 mm Intersil HPLC Flow rate 0.7 mL/min Temperature 35 C. Injection volume 10 L Detector DAD (Diode Array Detector)

(63) After HPLC analysis, a calibration curve was prepared with the content of vitamin C (g/L) as the abscissa axis and the area of the chromatogram as the ordinate axis. The areas of vitamin C of test solutions of Experimental example 4 and Comparative example 4 were read by using the following equation, and the content of vitamin C was calculated from the calibration curve.

(64) $\begin{array}{cc}\mathrm{Vitamin}C\mathrm{content}\left(\mathrm{mg}\text{/}100g\mathrm{soaked}\mathrm{solution}\right)=\frac{\begin{array}{c}\mathrm{Concentration}\mathrm{obtained}\mathrm{from}\mathrm{calibration}\mathrm{curve}\left(g\text{/}L\right)\\ \mathrm{dilution}\mathrm{volume}\left(\mathrm{mL}\right)1001000\end{array}}{\mathrm{Collected}\mathrm{sample}\mathrm{amount}\left(g\right)1000}& \left[\mathrm{Equation}2\right]\end{array}$

(65) As a result, it was found that the vitamin C content of Experimental example 4 and Comparative example 4 was not significantly different (Table 12). As a result, it could be found out that when the lemon-soaked solution was prepared using tagatose, the soaked solution of the same level, in terms of useful components, as the lemon-soaked solution using sucrose was embodied, too.

(66) TABLE-US-00012 TABLE 12 Vitamin C (mg/100 g soaked Classification solution) Comparative example 4 30.5 Experimental example 4 29.6