T∕CQAP 2001-2022 纳豆粉

1、学兔兔标准下载学兔兔标准下载ICS 67.060 CCS X 11准T/CQAP 200120222022-01-26 发布Natto powder2022-03-01 实施中国医药质量管理协会发T/CQAP 20012022学兔兔标准下载T/CQAP 20012022学兔兔标准下载目 次 TOC o 1-5 h z tuW I弓iw ni翻i2规范性引用文件1术语和定义1 2原辅料2 2生产加工过程中的卫生要求3检验方法3感官要求检验3理化指标检验3纳豆激酶活性检验 4微生物指标检验 4净含量4检验规则4出厂检验4型式检验4组批与抽样5判定原则5标签、标志、包装、运输、贮存和保质期5标签、标

2、志5 5瑜57.4贝亡# 5保质期5附录A（规范性）纳豆激酶测定方法紫外分光光度法 6附录B（规范性）纳豆激酶测定方法琼脂糖纤维蛋白平板法9参考t献12T/CQAP 20012022 学兔兔标准下载(B.4.11)，put it in a 100 mL beaker, heat it in a water bath at 37 C for 5 min, add agarose and thrombin solution, mix it for 10 s immediately, quickly pour it into the Petri dish and place it at room t

3、emperature for 1 h.Draw a circle with the same diameter of 14 cm at the bottom of the Petri dish with paper, draw a small circle with a diameter of 24 mm at the center of the circle, take the center of the small circle as the center point, and place the prepared plate. Punch vertically on the fibrin

4、 plate.The number of holes to be punched is the sum of the number of standard series plus the number of samples (including parallel samples). A distance of 15 mm shall be maintained between holes to prevent the intersection of dissolving rings and affect the diameter results of measurement.B.5.3 Det

5、erminationB.5.3.1 Urokinase control curve samplingAccurately measure 10 urokinase control solutions of different concentrations in table B.1 with a pipette. Place the vertical points in the fibrin plate holes prepared in B.5.2, and mark the concentration of each point.B.5.3.2 Natto powder sample sam

6、plingAccording to the estimated nattokinase content of the sample in advance (refer to the previous data)，weigh an appropriate amount of the sample into an appropriate volumetric flask, and the sample volume should make the final sample concentration between 200 lU/mL and 400 lU/mL. Accurately measu

7、re the sample solution 10 with a pipette point sample and make 1 2 parallel samples at the same time. The urokinase standard curve series and the sample solution shall be sampled at the midpoint of the same fibrin plate, and the sample number shall be accurately marked. After sampling ,put the plate

8、 cover on the back cover and place it in a 37 C incubator for reaction for 18 h.B.5.3.3 Solusphere measurementTake out the plate and immediately start to measure the diameter of the dissolving ring and calculate the area of the dissolving ring = kR2. Taking the logarithm of lysosomal area as the abs

9、cissa and the logarithm of concentration as the ordinate as the regression curve, the corresponding regression equation is obtained. The content C of nattokinase in the sample was calculated according to the regression equation.NOTE The ring dissolving reaction is still in progress after the plate i

10、s taken out. If the measurement time is delayed for a long time, it will affect the judgment of the ring diameter. Putting the plate into the refrigerator at 5 C can help the coil dissolving reaction stop temporarily.B.6 Result calculationThe Nattokinase Activity X (lU/g) in the sample is calculated

11、 according to formula (B.1)：(B.1 )X= Cx V/MwhereC is nattokinase in sample solution calculated by regression equation, lU/mL；V is total volume of sample dilution, mL；M is sample quality, g.B.7 Key control points of analysis stepsAttention shall be paid to the following operations when using this met

12、hod.Before the experiment, the plates and other instruments used shall be disinfected under the ultraviolet lamp for 30 min.Phosphate buffer and 0.9% sodium chloride solution shall be sterilized.Thrombin can be prepared into 20 U/mL or 20 BP/mL solution, sub packed in a small centrifuge tube and sto

13、red in a - 20 C refrigerator. It can be diluted according to the amount of fibrinogen before use.During the preparation of the plate, the plate shall be placed horizontally on the flat desktop, the flatness of the desktop shall be strictly checked by the spirit level, and bubbles shall be prevented

14、when pouring into the plate.During sample preparation, the sample weight can be directly estimated by referring to the approximate content of nattokinase in the sample, so that the final sample concentration can be in the range of 200 lU/mL400 lU/mL.The culture time of fibrin plate after sampling sh

15、all be strictly controlled according to the standard.If it is too late to measure multiple plates at the same time, put them in the refrigerator at 5 C first to inhibit the growth of dissolving ring.T/CQAP 20012022学兔兔标准下载T/CQAP 20012022学兔兔标准下载BibliographyQuality standard of culture extract of natto

16、fungus Fermented soya (natto) extract (No. 53) 9 Japan health and Nutrition Food AssociationStatement of nattokinase Association in JapanCommission Implementing Decision (EU) 2017/115 of 20 January 2017 authorizing the placing on the market of fermented soybean extract as a novel food ingredient under Regulation (EC) No 258/97 of the European Parliament and of the Council (notified under document C (2017) 165) (Only the English text is authentic)National drug standards WSt-(X-052)-2001 Z-2010 LumbrokinaseGB