CN107941967B

CN107941967B - Fingerprint detection method of xiaokeling tablets and xiaokeling capsules

Info

Publication number: CN107941967B
Application number: CN201810023255.8A
Authority: CN
Inventors: 张涛; 朱雪妍; 张颖婷; 林燕翔; 张慧; 黄博; 姚力; 罗轶
Original assignee: Guangxi Institute For Food And Drug Control
Current assignee: Guangxi Institute For Food And Drug Control
Priority date: 2018-01-10
Filing date: 2018-01-10
Publication date: 2020-08-04
Anticipated expiration: 2038-01-10
Also published as: CN107941967A

Abstract

The invention discloses a fingerprint detection method of xiaokeling tablets and xiaokeling capsules, which adopts a high performance liquid fingerprint detection method to efficiently detect characteristic peaks of various components such as moutan bark, medlar, astragalus, rehmannia root, coptis in the xiaokeling tablets and the xiaokeling capsules so as to judge whether enterprises have behaviors of no feeding or less feeding and feeding of inferior medicine materials, and can better control the feeding quantity and quality.

Description

Fingerprint detection method of xiaokeling tablets and xiaokeling capsules

Technical Field

The invention relates to the technical field of detection of traditional Chinese medicine preparations, in particular to a fingerprint detection method of xiaokeling tablets and xiaokeling capsules.

Background

The preparation method comprises decocting five materials including rehmanniae radix, radix Ophiopogonis, fructus Schisandrae chinensis, radix astragali, cortex moutan, Coptidis rhizoma, Poria, Ginseng radix Rubri, Trichosanthis radix, fructus Lycii and Gypsum Fibrosum in water for 2 times, concentrating into soft extract, mixing with the rest fine powders, drying, mixing with other adjuvants, granulating, and making into tablet or capsule. Has the effects of supplementing qi and nourishing yin, clearing heat and purging fire, promoting the production of body fluid and quenching thirst.

The existing standard mainly comprises the first part of 'Chinese pharmacopoeia' 2015 edition and the standard YBZ06282004 of the national food and drug administration, wherein microscopic and thin-layer chromatography identification is adopted in the standard to control only poria cocos, gypsum, coptis chinensis, wolfberry and other medicinal materials, and a plurality of medicinal materials lack effective control indexes.

Disclosure of Invention

The invention aims to provide a fingerprint detection method for xiaokeling tablets and xiaokeling capsules, which adopts a high performance liquid fingerprint detection method to efficiently detect characteristic peaks of various components such as moutan bark, medlar, astragalus, rehmannia root, coptis in the xiaokeling tablets and the xiaokeling capsules so as to judge whether enterprises have behaviors of no feeding or less feeding and feeding of inferior medicine materials, and can better control the feeding quantity and quality.

The fingerprint detection method of the diabetes tablet and the diabetes capsule comprises the following steps:

(1) chromatographic conditions and system applicability test, i.e., HICHROM Alltima C18 (250mm × 4.6.6 mm, 5 μm), acetonitrile as a mobile phase A, 0.01 mol/L monopotassium phosphate (PH adjusted to 3 by phosphoric acid) as a mobile phase B, gradient elution is carried out according to the table 1, the detection wavelength is 240nm, and the number of theoretical plates is not less than 10000 calculated according to paeoniflorin peaks.

TABLE 1 gradient elution Table

(2) The preparation of the test solution comprises taking a proper amount of sample, grinding, taking about 3g, precisely weighing, placing in a conical flask, precisely adding 100m L of methanol, weighing, heating and refluxing for 60 min, cooling, weighing again, supplementing the lost weight with methanol, shaking up, filtering, precisely absorbing the subsequent filtrate 50m L, concentrating under reduced pressure to dryness, diluting with 90% methanol to volume of 10m L, shaking up, and filtering.

(3) The control solution is prepared by precisely weighing appropriate amount of penoniflorin control, and adding methanol to obtain solution containing penoniflorin 30 μ g per 1m L.

(4) Establishing a comparison fingerprint.

(5) Methodological tests were performed, which included the following:

1) determining the attribution of chromatographic peaks;

2) testing the stability of the tested solution;

3) testing the precision of the instrument;

4) and (5) performing repeatability test.

(6) And (3) sample determination, namely precisely absorbing 10 mu L of each of the reference solution and the test solution according to chromatographic conditions, injecting the reference solution and the test solution into a liquid chromatograph, recording a 35-minute chromatogram, removing the chromatographic peak in the first 4 minutes, wherein the test chromatogram is basically consistent with the reference fingerprint and has 8 corresponding characteristic peaks, and the characteristic peak is an S peak relative to paeoniflorin.

The invention has the beneficial effects that:

the invention adopts a high performance liquid fingerprint detection method, can efficiently detect characteristic peaks of various components such as moutan bark, medlar, astragalus, rehmannia root, coptis in the diabetes tablet and the diabetes capsule, so as to judge whether the enterprise has the behavior of no feeding or less feeding and feeding of inferior medicine, and can better control the feeding quantity and quality.

Drawings

FIG. 1 is a high performance liquid chromatography fingerprint of lot ZKA1603 sample of company A;

FIG. 2 is a chromatogram of Xiaokeling tablet sample and rehmannia root;

FIG. 3 is a chromatogram of Xiaokeling tablet sample and fructus Lycii;

FIG. 4 is a chromatogram of Xiaokeling tablet sample and radix astragali;

FIG. 5 is an overlay of all manufacturer samples.

Detailed Description

In order to describe the present invention in more detail, the present invention will be further described with reference to the following examples.

Example 1

A fingerprint detection method for XIAOKELING tablet and XIAOKELING Capsule comprises the following steps:

instrument and reagent

Agilent 1260 high performance liquid chromatograph equipped with DAD detector, model M L204 electronic analytical balance (Metler-Tollido instruments Shanghai Co., Ltd.), Hei-VAP type rotary evaporator (Heidolph, Germany).

Paeoniflorin (provided by China institute for testing and testing food and drug, batch number: 110736-201741, content 95.7%);

acetonitrile and potassium dihydrogen phosphate reagents are chromatographically pure, and other reagents are analytically pure;

the samples of the xiaokeling tablets and the xiaokeling capsules are samples of national evaluative spot tests in 2017 years;

the diabetes tablet used in the methodology test is company A, batch number: ZKA 1603.

The detection method comprises the following steps:

TABLE 1 gradient elution Table

(4) Establishing a contrast fingerprint spectrum: the sample of company A has more batches in the sampling, and 111 batches in total indicate that the sample has larger market share; the standard used by the sample of the company B is a national food and drug administration standard YBZ06282004, the standard collects the content determination of berberine hydrochloride, the control index is more than that of the current pharmacopoeia, and the experiment shows that the quality of the samples of the two enterprises is stable, so the samples of the two enterprises are generated by contrasting fingerprint spectrums. See fig. 1.

(5) Methodological tests were performed, which included the following:

1) chromatographic peak assignment

Comparing the chromatogram of XIAOKELING tablet with chromatogram of rehmanniae radix, fructus Lycii and radix astragali respectively, as shown in fig. 2-4, and the chromatogram peaks are shown in table 2.

TABLE 2 assignment table of various spectral peaks of Xiaokeling tablets

2) Stability test of the tested solutions: taking the same sample solution (ZKA 1603 of A company), measuring the sample solution once at regular intervals according to chromatographic conditions, investigating for 24 hours totally, measuring for 7 times, introducing the measured spectrum into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system recommended by the national pharmacopoeia, taking a paeoniflorin peak as a reference peak, calculating by software, and obtaining the result that the average similarity between the fingerprint of the sample solution and the fingerprint of a reference substance is 0.990 and the RSD is 0.49% (n is 7) in 7 times of measurement, which is shown in Table 3. The test shows that the test solution is stable for at least 24 hours.

Table 3 stability test results

3) Precision test of instrument

The same sample solution (ZKA 1603, company A) was taken and subjected to 6-time continuous measurement under chromatographic conditions. The measured spectrum is introduced into a traditional Chinese medicine chromatogram fingerprint similarity evaluation system recommended by the national pharmacopoeia, the paeoniflorin peak is taken as a reference peak, the software calculates, and the average similarity of the fingerprint of the sample to be measured and the fingerprint of the reference substance is 0.989 and RSD is 0.20 percent (n is 6) in 6 times of measurement results, which is shown in table 4. The test shows that the instrument precision of the method is good.

TABLE 4 results of precision test

4) Repeatability test

Taking the same sample solution (ZKA 1603 of A company), parallelly measuring 6 parts according to chromatographic conditions, introducing the measured spectrum into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system recommended by the national pharmacopoeia, taking a paeoniflorin peak as a reference peak, calculating by software, and obtaining the result that the average similarity between the sample spectrum and the reference fingerprint is 0.991 and the RSD is 0.46% (n is 6) after 6 times of measurement, which is shown in Table 5. Test results show that the method has good reproducibility.

TABLE 5 results of the repeatability tests

(6) The sample determination comprises precisely absorbing 10 mu L of a reference solution and a test solution respectively according to chromatographic conditions, injecting into a liquid chromatograph, recording 35-minute chromatogram, removing the chromatogram peak of the previous 4 minutes, wherein the chromatogram of the test solution is basically consistent with the reference fingerprint, has 8 corresponding characteristic peaks, and is an S peak relative to paeoniflorin, determining the fingerprint of 111 batches of xiaokeling tablets and two batches of xiaokeling capsules of different manufacturers, summarizing all samples, and showing the results in tables 6-8, and showing the overlapped graph of the samples of each manufacturer in figure 4.

TABLE 6 summary of similarity of various manufacturers

TABLE 7 Xiaokeling capsule similarity results

TABLE 8 summary of fingerprint disqualification results for all sampled samples

Claims

1. The fingerprint detection method of the xiaokeling tablets and the xiaokeling capsules is characterized by comprising the following steps:

(1) chromatographic conditions and system applicability tests comprise that the HICHROM Altima C18 takes acetonitrile as a mobile phase A and 0.01 mol/L monopotassium phosphate as a mobile phase B, wherein the pH of the 0.01 mol/L monopotassium phosphate is adjusted to 3 by phosphoric acid, gradient elution is carried out for 57 minutes, the elution procedure is 0-16 min, 5% A → 23% A, 16-35 min, 23% A → 30% A, 35-50 min, 30% A → 48% A, 50-51 min, 48% A → 5% A, 51-57 min, 5% A, the detection wavelength is 240nm, and the number of theoretical plates is not less than 10000 calculated according to paeoniflorin peaks;

(2) preparing a test solution, namely taking a proper amount of samples, grinding, taking 3g of the samples, precisely weighing, placing the samples in a conical flask, precisely adding 100m L of methanol, weighing, heating and refluxing for 60 minutes, cooling, weighing again, supplementing the lost weight with the methanol, shaking up, filtering, precisely absorbing 50m L of a subsequent filtrate, concentrating the filtrate under reduced pressure to dryness, fixing the volume to 10m L with 90% of methanol, shaking up, and filtering to obtain the test solution;

(3) preparing control solution by precisely weighing appropriate amount of penoniflorin control, and adding methanol to obtain solution containing penoniflorin 30 μ g per 1m L;

(4) establishing a comparison fingerprint spectrum;

(5) methodological tests were performed, which included the following:

1) determining the attribution of chromatographic peaks: comparing the chromatogram of XIAOKELING tablet with the chromatogram of rehmanniae radix, fructus Lycii and radix astragali respectively;

2) testing the stability of the tested solution;

3) testing the precision of the instrument;

4) performing a repeatability test;

(6) and (3) sample determination, namely precisely absorbing 10 mu L of the reference solution and the test solution respectively according to chromatographic conditions, injecting the reference solution and the test solution into a liquid chromatograph, recording a 35-minute chromatogram, removing the chromatographic peak in the first 4 minutes, wherein the test chromatogram is basically consistent with the reference fingerprint and has 8 corresponding characteristic peaks, and the characteristic peak is an S peak relative to paeoniflorin.