CN104049047B - A kind of method evaluating quality of stevioside - Google Patents
A kind of method evaluating quality of stevioside Download PDFInfo
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Abstract
The invention provides the new method that a kind of quality of stevioside is evaluated, the method comprises following process: adopt 9 kinds of major glycosides compositions in chromatography or other analytical approach Accurate Calibration steviosides, each glycoside component proportion in stevioside is multiplied by its corresponding sugariness multiple, results added is the sugariness multiple of stevioside, combines according to stevioside assay result the quality that the sugariness multiple calculating gained judges stevioside.The method and traditional employing are tasted method and are demarcated stevioside sugariness and have more objectivity, authenticity and accuracy, accurately can distinguish the quality discrepancy of identical or different serial stevioside.
Description
Technical field
The present invention relates to a kind of method evaluating quality of stevioside, be specifically related to the evaluation method adopting stevioside sugariness objective quantification quality of stevioside.
Background technology
Stevioside is a kind of new type natural sweetener extracted from composite family herbaceous wild plants STEVIA REBAUDIANA, is to continue outside sugarcane, beet sugar that the third has Development volue and healthy natural sweetener of praising highly, and is described as in the world in " sugar source, third place in the world ".It has the feature of high, the high sugariness of security, low calorie.
Stevioside originates in the ground such as South America Paraguay, Brazil, and the edible existing centuries history of local resident, find no any murder by poisoning so far, prove through domestic and international pharmacological evaluation, steviol glycoside is non-carcinogenic material, no side effects, edible safety.Its sugariness is 200 to 400 times of sucrose, and heat is only 1/300 of sucrose, can be applicable to low-calorie diet, beverage and medicine.Steviol glycoside participates in metabolism hardly, only can feel its sweet taste after taking, and major part is discharged from urine and ight soil with original shape.Long-term edible steviol glycoside can not make people get fat, and has the function of hypoglycemic, anti-caries, stomach invigorating, relieving fatigue, and has auxiliary curative effect to edible for patients such as body building and weight reducing and diabetes, heart disease, hypertension, artery sclerosis.
In recent years the research of STEVIA REBAUDIANA chemical composition is found, the chemical composition contained in STEVIA REBAUDIANA comprises steviol glycoside (stevioside), steviolbioside (steviolbioside), Rebaudiodside A (rebaudioside) A, B, C, D, E, F, Du Ke glycosides (dulcoside) A, B, the glucosides constituents such as Rubusoside (rubusoside).In addition, also containing sterols: there is stigmasterol (stigmasterol), cupreol (β-sitosterol), campesterol (campesterol), stigmasterol-β-D-Glucose glycosides (stigmasterol-β-D-gluco-side), cupreol-β-D-Glucose glycosides (β-sitosterol-β-D-glucoside), another containing flavonoids and methods of glycosides thereof: api-genin-7-O-β-D-glucoside (apigenin-7-O-β-D-glucoside), apiolin-4-O-glucoside (apigenin-4-O-gluco-side), cyanidenon-7-O-glucoside (luteolin-7-O-glucoside), Kaempferol-O-rhamnoside (daempferol-3-O-rhamnoside), quercitin (quercetrin), Quercitrin-3-O-glucoside (quercetin-3-O-glucoside), Quercetin-3-O-Arabinoside (quercetin-3-O-arabinoside), 5, 7, 3-trihydroxy-3, 6, 4-trimethoxy flavone (5, 7, 3-trihydroxy-3, 6, 4-trimethoxy flavone).
Stevioside is as a kind of emerging natural sweetener, and its sugariness is mainly derived from the glucosides such as steviol glycoside and Rebaudiodside A constituents.According to JECFA(Joint FAO/WHO Expert Committee on Food Additives, the food additives joint specialist council under FAO (Food and Agriculture Organization of the United Nation) and the World Health Organization (WHO)) the stevioside quality standard of working out, the quality of commodity stevioside realizes mainly through the 9 kinds of principal ingredients controlled in stevioside, these 9 kinds of compositions comprise steviol glycoside (stevioside), steviolbioside (steviolbioside), Rebaudiodside A (rebaudioside) A, B, C, D, F, Du Ke glycosides (dulcoside) A and Rubusoside (rubusoside).The composition of stevioside owing to being multiple glucosides constituents, so there is the situation that series is many, combination is many in commodity stevioside.The mainstream product in the market comprises R-A series (Reb-A 40%, Reb-A 50%, Reb-A 60%, Reb-A 80%, Reb-A 90%, Reb-A 95%, Reb-A 97%, Reb-A 98%, Reb-A 99%), STV series (STV 90%, STV 95%, STV 98%), common stevioside series (total steviol glycoside Stevia 80%, Stevia 85%, Stevia 90%, Stevia 95%), except highly purified Reb-A and STV, all the other products are the composition of multiple glucosides, and each glucosides ratio in the composition has uncertainty.Particularly common stevioside series, it only requires that the total glycosides of stevioside (TSG) reaches certain content, and does not do requirement, so composition has larger uncertainty to each glycosides content in the product and ratio.The total glycosides of stevioside (TSG) series of products, owing to having higher cost performance, have become stevioside product most popular on market at present.9 kinds of glucosides that in JECFA standard, defined will be tested, sugariness fold difference is very big, the rebaudioside A that sugariness is the highest is 400 times of sweetness of cane sugar, and the sugariness of steviolbioside is only about 10 times of sucrose, even if so the total glycosides of stevioside (TSG) reaches certain content, the sugariness multiple of product still likely has larger difference, and commodity stevioside finally uses as sweetener, and sugariness determines the objective standard of its consumption instead of the content of the total glycosides of stevioside (TSG).Realize mainly through tasting the demarcation of stevioside sugariness at present, due to the individual differences of tester and the mouthfeel of stevioside composite complexity, the sugariness wanting to taste by individual Accurate Calibration stevioside has certain difficulty, and does not have objectivity.Current urgent need develops new quality of stevioside evaluation method, the inherent quality of stevioside can be reflected objective reality, need when particularly it uses as sweetener to demarcate its sugariness objectively, and carry out the quality of comprehensive evaluation stevioside in conjunction with the content of the total glycosides of stevioside (TSG).
Summary of the invention
The present invention measures the content of 9 kinds of glucosides in stevioside by chromatography or other analytical approachs, the percentage composition of each glucosides is multiplied by its sugariness multiple respectively, the results added obtained is total sugariness of stevioside, evaluated the quality of stevioside by total glycosides content of stevioside and sugariness, this evaluation method mainly comprises the following steps:
Step one: the assay of 9 kinds of major glycosides in stevioside;
According to the examination criteria of JECFA, main mensuration 9 kinds of glucosides in stevioside, be respectively steviol glycoside (stevioside), steviolbioside (steviolbioside), Rebaudiodside A (rebaudioside) A, B, C, D, F, Du Ke glycosides (dulcoside) A and Rubusoside (rubusoside), the structural formula of each glucosides is shown in accompanying drawing 1.
The assay of 9 kinds of glucosides can adopt high performance liquid chromatography (HPLC), vapor-phase chromatography (GC), capillary electrophoresis (CE) isochromatic spectrum analytical approach or other analytical approachs.JECFA adopts high performance liquid chromatography (HPLC) to demarcate the content of 9 kinds of glucosides, and chromatographic condition is as follows:
Chromatographic column: C18 post (250 × 4.6 mm, 5 μm)
Mobile phase: acetonitrile-10mmol/L sodium phosphate (32:68), pH 2.6
Flow velocity: 1ml/min
Determined wavelength: 210 nm
Column temperature: 40
oc
Gained standard diagram is shown in accompanying drawing 2.
Step 2: the calculating of stevioside sugariness multiple;
In stevioside, the sugariness multiple of 9 kinds of major glycosides is in table 1, and the percentage composition shared in stevioside according to each glucosides measured in " 1 ", be multiplied by its corresponding sugariness multiple, results added is the sugariness multiple of stevioside.
Stevioside sugariness multiple=STV% × 300+(R-A) % × 400+(R-B) % × 10+(R-C) % × 50+(R-D) % × 200+(R-F) % × 50+(S-Bio) % × 10+(D-A) % × 50+(R-S) % × 150
The sugariness multiple of 9 kinds of major glycosides in table 1 stevioside
Step 3: the quality judging stevioside product according to total glycosides content of stevioside and sugariness multiple, when the total glycosides content of stevioside reaches requirement, the product quality that sugariness multiple is high is better.
Accompanying drawing explanation
Fig. 1 shows the chemical structural formula of 9 kinds of glucosides in stevioside.
Fig. 2 shows the standard diagram that JECFA adopts 9 kinds of glucosides in high-performance liquid chromatogram determination stevioside.
Fig. 3 shows the method according to evaluation quality of stevioside of the present invention.
Embodiment
Describe the present invention below in conjunction with specific embodiment, but be not limited in following instance.
Embodiment 1:
Total glycosides 90 stevioside sample 1
Get stevioside sample 1, by JECFA standard detection, chromatographic condition is as follows:
Chromatographic column: C18 post (250 × 4.6 mm, 5 μm)
Mobile phase: acetonitrile-10mmol/L sodium phosphate (32:68), pH 2.6
Flow velocity: 1ml/min
Determined wavelength: 210 nm
Column temperature: 40
oc
Assay the results are shown in Table 2.
The content of 9 kinds of glucosides in the total glycosides 90 stevioside sample 1 of table 2
The total glycosides content=64.2%+15.9%+5.8%+0.8%+0.6%+1.1%+1.5%+1.3%=91.2% of stevioside
Stevioside sugariness multiple=64.2% × 300+15.9% × 400+5.8% × 50+0.8% × 200+0.6% × 50
+1.1%×10+1.5%×50+1.3%×150=263.8
Embodiment 2:
Total glycosides 90 stevioside sample 2
Content assaying method, with example 1, the results are shown in Table 3.
The content of 9 kinds of glucosides in the total glycosides 90 stevioside sample 2 of table 3
The total glycosides content=22.9%+60.4%+0.8%+3.5%+1.2%+0.3%+0.5%+1.3%=90.9% of stevioside
Stevioside sugariness multiple=22.9% × 300+60.4% × 400+0.8% × 10+3.5% × 50+1.2% × 200+0.3% ×
10+0.5%×50+1.3%×150=316.8
Embodiment 3:
Total glycosides 95 stevioside sample
Content assaying method, with example 1, the results are shown in Table 4.
The content of 9 kinds of glucosides in the total glycosides 95 stevioside sample of table 4
The total glycosides content=50.8%+28.3%+1.1%+12.5%+1.3%+0.2%+0.5%+0.7%=95.4% of stevioside
Stevioside sugariness multiple=50.8% × 300+28.3% × 400+1.1% × 10+12.5% × 50+1.3% × 200+0.2% ×
50+0.5%×50+0.7%×150=276
The total glycosides of stevioside 90 and total glycosides 95 series are owing to having higher cost performance, and become the most popular product in market at present, the price of the two has larger difference.Embodiment 1 and embodiment 2 show, though two kinds of steviosides are same specification (being total glycosides 90), sugariness differs greatly, and embodiment 2 and embodiment 3 show, high standard product (total glycosides 95) may not be higher than the sugariness of low specification product (total glycosides 90).By the requirement of GB GB-8270-1999 to sugariness, sugariness be greater than 250 be superfine stevioside, 3 kinds of stevioside products all meet the requirements, and adopt trial test method cannot distinguish their sugariness difference accurately, objectively.
The invention provides a kind of method that accurately can indicate stevioside sugariness, and in conjunction with the total glycosides measurement result of stevioside in assay, evaluate the quality of stevioside, for the use business of stevioside and consumer provide a kind of method selecting the applicable stevioside oneself used.
Claims (1)
1. evaluate a method for quality of stevioside, it is characterized in that, combined by total glycosides content of stevioside and sugariness and evaluate the quality of stevioside, said method comprising the steps of:
Step one, employing chromatogram analysis method measure the percentage composition of 9 kinds of major glycosides in stevioside, wherein, described 9 kinds of major glycosides are respectively steviol glycoside, steviolbioside, rebaudioside A, B, C, D, F, Du Ke glycosides A and Rubusoside, described chromatogram analysis method is high performance liquid chromatography, vapor-phase chromatography, capillary electrophoresis, wherein, the chromatographic condition of high performance liquid chromatography is as follows:
Chromatographic column: C18 post, 250 × 4.6 mm, 5 μm
Mobile phase: acetonitrile-10mmol/L sodium phosphate, 32:68, pH2.6
Flow velocity: 1ml/min
Determined wavelength: 210nm
Column temperature: 40 ° of C;
Step 2, calculating stevioside sugariness multiple, the percentage composition shared in stevioside according to each major glycosides of step one mensuration, be multiplied by its corresponding sugariness multiple, results added is the sugariness multiple of stevioside, wherein, the sugariness multiple of described each major glycosides is respectively:
Steviol glycoside (STV): 300
Lay bud enlightening glycosides A, (R-A): 400
Lay bud enlightening glycosides B, (R-B): 10
Lay bud enlightening glycosides C, (R-C): 50
Lay bud enlightening glycosides D, (R-D): 200
Lay bud enlightening glycosides F, (R-F): 50
Steviolbioside, (S-Bio): 10
Du Ke glycosides A, (D-A): 50
Rubusoside, R-S:150
So, the sugariness multiple=STV% × 300+(R-A of stevioside) % × 400+(R-B) % × 10+(R-C) % × 50+(R-D) % × 200+(R-F) % × 50+(S-Bio) % × 10+(D-A) % × 50+(R-S) % × 150;
Step 3, judge the quality of stevioside product according to total glycosides content of stevioside and sugariness multiple, when the total glycosides content of stevioside reaches requirement, the product quality that sugariness multiple is high is better.
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| CN104483424B (en) * | 2014-12-18 | 2017-01-18 | 广西壮族自治区中医药研究院 | Rubusoside quality control method |
| CN106814143A (en) * | 2015-12-01 | 2017-06-09 | 桂林三金药业股份有限公司 | A kind of Steviosin quality determining method |
| CN106282271A (en) * | 2016-09-12 | 2017-01-04 | 江南大学 | A kind of method applying at alternating temperature catalysis Lay bud enlightening glycosides B to prepare Lay bud enlightening glycosides D |
| CN108732278B (en) * | 2018-08-01 | 2021-06-01 | 山东省食品药品检验研究院 | Method for simultaneously determining ten natural sweeteners in beverage by high performance liquid chromatography |
| CN111879865B (en) * | 2020-06-13 | 2021-11-05 | 武汉迈特维尔生物科技有限公司 | Method for separating and identifying stevioside |
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