A quality detection method for Helleborus officinalis medicinal material

Abstract

The invention discloses a quality detection method for Helleborus truncatus medicinal materials, which includes a method for determining the content of isofraxidin and scopoletin in Helleborus truncatus medicinal materials, and can accurately and quickly determine the content of Helleborus truncatus medicinal materials, and provide effective qualitative and quantitative analysis means for Helleborus truncatus medicinal materials, thereby facilitating the quality control of Helleborus truncatus medicinal materials. The quality detection method for Helleborus truncatus medicinal materials provided by the invention can effectively control the overall quality of Helleborus truncatus medicinal materials and ensure the safety and effectiveness of clinical use of Helleborus truncatus medicinal materials.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine analysis, and particularly relates to a quality detection method for Helleborus officinalis medicinal materials. Background Art

[0002] Iron chopsticks are a traditional medicinal material used by the Miao people. They are the dried stems, branches or fine roots of Chimonanthus nitens Oliv. or Chimonanthus praecox (Linn.) Link, plants of the Chimonanthus family. The 2003 edition of the Quality Standards for Chinese Medicinal Herbs and Ethnic Medicinal Materials in Guizhou Province includes the fine roots of these two species under the name of "iron chopsticks". They have the functions of dispelling wind and relieving pain, promoting blood circulation and detoxifying, and are used for asthma, cough caused by overwork, stomachache, rheumatic pain, sores, boils, swelling and bruises, etc. They are the raw materials for the preparations of Ganqing Capsules, Compound Xianling Rheumatism Wine, Compound Xueteng Wine, Liangjiang Weiyang Capsules, Qingbi Tongluo Wine, Shenglong Qufeng Wine, Tongluo Guzhining Paste, etc.

[0003] The original standard only stipulated [properties] and [identification] (cross section of roots, thin layer identification of control medicinal materials), and the standard was not perfect. Through literature survey and market survey, commercial medicinal materials samples of Helleborus thibetanus were collected, and it was clarified that the main medicinal parts of Helleborus thibetanus are stems and branches. According to the research results, the quality standard of Helleborus thibetanus was improved. Different from the "Helleborus thibetanus" included in the "Shaanxi Provincial Medicinal Materials Standard" (2015 edition), it is the dried root and rhizome of the Ranunculaceae plant Helleborus thibetanus Franch.

[0004] The original standard did not establish content determination items, and could not effectively control the overall quality of Helleborus truncatus medicinal materials. Therefore, the development and establishment of a quality detection method for Helleborus truncatus is of great significance to control the overall quality of Helleborus truncatus medicinal materials and ensure the safety and effectiveness of clinical use of Helleborus truncatus medicinal materials. Summary of the invention

[0005] The purpose of the present invention is to provide a quality detection method for Helleborus helix medicinal materials in view of the deficiencies of the prior art. The quality detection method includes a method for determining the content of isofraxidin and scopolamine in Helleborus helix medicinal materials. The quality detection method can accurately identify the authenticity of the medicinal materials, and can accurately and quickly determine the content of isofraxidin and scopolamine in Helleborus helix medicinal materials, thereby facilitating the quality control of Helleborus helix medicinal materials. The quality detection method of Helleborus helix medicinal materials can effectively control the overall quality of Helleborus helix medicinal materials and ensure the safety and effectiveness of clinical use of Helleborus helix medicinal materials.

[0006] To achieve the above object, the present invention adopts the following technical solution:

[0007] The invention discloses a quality detection method for a Helleborus helveticus medicinal material. The content determination is to determine the contents of isofraxidin and scopoletin in the Helleborus helveticus medicinal material by high performance liquid chromatography.

[0008] The method for determining the content of isofraxidin and scopolamine in the medicinal material of Helleborus officinalis is determined by high performance liquid chromatography, comprising the following steps:

[0009] (1) Chromatographic conditions and system suitability test: Chromatographic column: Cosmosil C 18 Column (4.6 mm×250 mm, 5 μm); mobile phase: acetonitrile: 0.1% formic acid water (14:86); detection wavelength: 341 nm; flow rate: 1 ml / min; column temperature set to room temperature; injection volume: 10 μl.

[0010] (2) Preparation of reference solution: Take appropriate amount of scopoletin and isofraxidin reference substances, weigh accurately, and add methanol to prepare a mixed solution containing 0.066 mg of scopoletin and 0.065 mg of isofraxidin per 1 ml.

[0011] (3) Preparation of test solution: Take about 1.0 g of the product powder (passed through a No. 3 sieve), accurately weigh it, place it in a stoppered conical flask, accurately add 40 ml of 80% methanol, weigh it, heat and reflux for 1.5 hours, cool, weigh it again, make up the lost weight with 80% methanol, shake well, filter, and take the filtrate to obtain the solution.

[0012] (4) Determination method: Accurately pipette 10 μl of the reference solution and the test solution respectively, inject into the liquid chromatograph, and determine;

[0013] (5) This product contains isoflurane (C 11 H 10 O5) shall not be less than 0.12%, scopoletin (C 10 H8O4) shall not be less than 0.20%.

[0014] The beneficial effects of the present invention are:

[0015] 1. The present invention provides a quality detection method for Helleborus truncatus medicinal materials, which includes a method for determining the content of isofraxidin and scopoletin in Helleborus truncatus medicinal materials, and can accurately and quickly determine the content of Helleborus truncatus medicinal materials, providing effective qualitative and quantitative analysis methods for Helleborus truncatus medicinal materials, thereby facilitating the quality control of Helleborus truncatus medicinal materials. The quality detection method for Helleborus truncatus medicinal materials provided by the present invention can effectively control the overall quality of Helleborus truncatus medicinal materials and ensure the safety and effectiveness of clinical use of Helleborus truncatus medicinal materials.

[0016] The quality detection method of the Helleborus officinalis medicinal material provided by the present invention helps to make up for the deficiencies of the quality detection method of the "Quality Standards for Chinese Medicinal Materials and Ethnic Medicinal Materials in Guizhou" (2003 Edition: Helleborus officinalis / Scolopendra subspinipes), and has strong practicality. The method of the present invention is of great significance for further quality control of Helleborus officinalis medicinal materials, and can provide a scientific basis for the formulation of the quality table of Helleborus officinalis medicinal materials. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 HPLC profiles were investigated for methanol solvent extraction;

[0018] Figure 2 HPLC profiles were investigated for ethanol solvent extraction;

[0019] Figure 3 To investigate the HPLC profile for the ultrasonic extraction method;

[0020] Figure 4 Investigate HPLC profiles for reflux extraction methods;

[0021] Figure 5 To investigate the HPLC profile for the cold infusion extraction method;

[0022] Figure 6 The HPLC spectrum was investigated for 60% methanol concentration extraction;

[0023] Figure 7 HPLC spectra were investigated for extraction with 80% different methanol concentrations;

[0024] Figure 8 The HPLC spectra were investigated for 100% extraction with different methanol concentrations;

[0025] Fig. 9 The HPLC profile was investigated for 30 min extraction time;

[0026] Fig.10 The HPLC profile was investigated for 1 h extraction time;

[0027] Fig.11 The HPLC profile was investigated for 1.5 h extraction time;

[0028] Fig.12 The HPLC profile was investigated for the 10 ml solid-liquid ratio;

[0029] Fig.13 The HPLC profile was investigated for the 20 ml solid-liquid ratio;

[0030] Fig.14 The HPLC profile was investigated for the 40 ml solid-liquid ratio;

[0031] Fig.15 The HPLC profile was investigated for the 60 ml solid-liquid ratio;

[0032] Fig.16 The UV absorption spectra of scopolamine and isofraxidin (A: isofraxidin; B: scopolamine);

[0033] Fig.17 The chromatograms of scopolamine and isofraxidin (A: scopolamine and isofraxidin mixed reference substance; B: Helleborus officinalis sample; C: blank solvent; 1: scopolamine; 2: isofraxidin);

[0034] Fig.18 This is the working curve of scopolamine and isofragidine. DETAILED DESCRIPTION

[0035] The present invention is further described below in conjunction with specific embodiments.

[0036] The method for detecting the quality of the medicinal material of Helleborus truncatus described in the present invention is not limited to the following embodiments.

[0037] Example

[0038] instrument:

[0039] High performance liquid chromatograph (Shimadzu CTO-20AC, Shimadzu Corporation, Japan); 1 / 100,000 electronic balance (model: AUW220D, Shimadzu Corporation); 1 / 10,000 electronic balance (model: BS224S, Sartorius Corporation); ultrasonic cleaning machine (model: KQ-4000B Gongyi Yuhua Instrument Co., Ltd.); chromatographic column (model: Amethyst C18-H, 4.6 mm × 250 mm, 5 μm); MiLLipore SimpLicity ultrapure water device (Merck Millipore); KQ5200 water bath (Kunshan Ultrasonic Instrument Co., Ltd.); DHG-9140A electric constant temperature blast drying oven (Gongyi Yuhua Instrument Co., Ltd.); 8-10 box-type resistance furnace (manufactured by Shenyang Energy Saving Electric Furnace Factory, China); Sony 717 digital camera (Sony Corporation, Japan); silica gel G plate (Qingdao Ocean Chemical).

[0040] Reagents and test drugs

[0041] 1. Reference

[0042] Scopoletin (batch number: BCTG-0159, purity>98%) and isofraxidin (batch number: BCTG-0780, purity>98%) were purchased from Jiangxi Bencao Tiangong Co., Ltd.

[0043] 2. Reagents

[0044] Methanol and acetonitrile (chromatographic grade, Fisher Scientific, Fairlawn, NJ, USA); formic acid (chromatographic grade, Sigma-Aldrich Co. Ltd, St Louis, MO, USA); methanol (analytical grade, Jinan Tianjiang Chemical Co., Ltd.); cyclohexane (analytical grade, Yucheng Chemical Plant of Dongyuwang Industrial Co., Ltd.); petroleum ether and toluene (Guangdong Shantou Xilong Chemical Co., Ltd.); ethyl acetate (analytical grade, Tongchuang Pharmaceutical Co., Ltd.); ethanol (analytical grade, Shanghai Zhenxing Chemical Plant No. 1); distilled water (Watson Group).

[0045] Medicinal Materials

[0046] The medicinal materials used in this experiment were provided by Guizhou Bailing Pharmaceutical, with a total of 20 batches. They were identified by Zhong Guoyue, a researcher at the Center for Chinese Medicine Resources and Ethnomedicine of Jiangxi University of Traditional Chinese Medicine, as the dried stems, branches and fine roots of Chimonanthus nitens Oliv. and Chimonanthus praecox (Linn.) Link, both of which belong to the Chimonanthus family. The stems are the main components, with fewer fine roots. The medicinal material information is shown in Table 1.

[0047] Table 1 Information on medicinal materials of Helleborus chinensis

[0048]

[0049] [Name of medicinal material] Iron chopsticks.

[0050] "Iron chopsticks" is a folk name of the Miao ethnic group. In the Miao language, it is called "Ghab jongx ghab link det ghab dliub (Gag Gonggale Dougatou)" (Southeast Guizhou). It is also known as Tiegangcha, Wa Niaochai, and Yanma Sang. The Buyi ethnic group also calls it "iron chopsticks" for medicinal purposes.

[0051] The "Helleborus thibetanus" listed in the Shaanxi Province Medicinal Materials Standard (2015 edition) is the dried root and rhizome of the Ranunculaceae plant Helleborus thibetanus Franch. It is different from the Miao medicine "Helleborus thibetanus".

[0052] [Source] Chimonanthus nitens Oliv. or Chimonanthus praecox (Linn.) Link, a plant of the Chimonaceae family, and its dried stems, branches and fine roots.

[0053] The original Guizhou local standard included the dried stems and fine roots of Chimonanthus nitens Oliv. and Chimonanthus praecox (Linn.) Link of the Chimonanthus family. It was identified by Zhong Guoyue, a researcher at the Center for Chinese Medicine Resources and Ethnic Medicine Research of Jiangxi University of Traditional Chinese Medicine, and was found to be consistent with the original standard. (Table 1)

[0054] Helleborus is an evergreen shrub, 1 to 3 meters high. The young branches are square, the old branches are nearly cylindrical, and are slightly hairy, gradually becoming hairless. The leaves are papery to nearly leathery, elliptical to ovate-lanceolate, 2 to 13 cm long, 1.5 to 5.5 cm wide, gradually pointed at the apex, obtuse to acute at the base, slightly rough and shiny on the leaf surface, with inconspicuous glandular hairs at the base, glabrous on the back of the leaf, or sometimes with short soft hairs on the leaf margins, veins and petioles; the veins are flat on the upper surface of the leaf, convex on the back of the leaf, and the reticulate veins are not obvious. The flowers are small, 7 to 10 mm in diameter, yellow or yellow-white; the tepals are round, ovate, obovate, ovate-lanceolate or oblong, 3 to 15 mm long, 2.5 to 10 mm wide, with short soft hairs on the outside and no hairs on the inside; the stamens are 2 mm long, the filaments are short, with short soft hairs, the anthers are ovate, curved inward, longer than the filaments, and the reduced stamens are 1.5 mm long; the carpels are 2 mm long, with sparse hard hairs at the base and the base of the style. The fruit is tantalum-shaped, 2 to 5 cm long, with a contracted mouth, gray-brown when mature, with short hairs, and aggregated achenes inside. The flowering period is from October to January of the following year, and the fruiting period is from April to July. It is distributed in Anhui, Zhejiang, Jiangsu, Jiangxi, Fujian, Hubei, Hunan, Guangxi, Yunnan, Guizhou, Shaanxi and other provinces and regions. It grows in mountain sparse forests or limestone mountains. The Miao, Buyi, Dong, Shui and other ethnic groups use it as medicine, with flowers, leaves, stems, branches and roots as medicine.

[0055] [Properties] The original Guizhou Provincial Standard stipulated the description of the properties of medicinal materials (fine roots). According to the investigation, the actual medicinal materials used are mainly stems and branches, so this standard has revised the description of the original standard properties according to the actual situation of the medicinal materials.

[0056] The stem of this product is cylindrical, with a diameter of up to 3cm, the outer skin is gray-brown, and lenticels are visible; the young branches are slightly square, waxy, hairless or sparsely hairy. The fine roots are curved and long cylindrical, with a diameter of 0.3-1cm, the outer skin is dark brown, rough, with obvious raised longitudinal wrinkles and round root marks. The texture is hard and not easy to break. The cross section of the cortex is brown, the wood is broad, light yellow-white, and has radial textures. The smell is fragrant, and the taste is spicy and slightly bitter.

[0057] [Content Determination] There is no provision for content determination in the original standard of iron chopsticks.

[0058] According to literature reports and the research in this revision, a total of 41 compounds mainly composed of coumarins were identified in the methanol extract of Helleborus truncatus, including 27 coumarins and their glycosides, 7 flavonoids, 3 terpenes, 3 alkaloids, 1 phenolic acid, and 1 other. Coumarin components are the main components of "Helleborus truncatus", and literature reports that scopoletin and isoflavone are the main active ingredients of Helleborus truncatus for asthma, cough caused by overwork, stomachache, abdominal pain, rheumatic arthralgia, furunculosis, and injuries caused by falls, which have a good correlation with the functions and indications of Helleborus truncatus.

[0059] In this revision, scopolamine and isofraxidin were selected as indicator components, and the HPLC content determination method was established. Based on the content determination results of 20 batches of samples, the limits were set. Finally, isofraxidin and scopolamine were determined as the indicator components for content determination, and the limits were set.

[0060] Determine by high performance liquid chromatography (General Rule 0512 of Part IV of the 2015 edition of the Chinese Pharmacopoeia). The determination method is as follows:

[0061] Preparation of test solution: Take about 1.0 g of the powder of the product (passed through No. 3 sieve), weigh it accurately, place it in a stoppered conical flask, add 40 ml of 80% methanol accurately, weigh it, heat and reflux for 1.5 hours, cool, weigh it again, make up the lost weight with 80% methanol, shake well, filter, and take the filtrate to obtain the solution.

[0062] Preparation of reference solution: Take appropriate amount of scopoletin and isofraxidin reference substances, weigh accurately, add methanol to prepare a mixed solution containing 0.066 mg of scopoletin and 0.065 mg of isofraxidin per 1 ml.

[0063] Chromatographic conditions: Column: Cosmosil C 18 Column (4.6 mm×250 mm, 5 μm); mobile phase: acetonitrile-0.1% formic acid water (14:86); detection wavelength: 341 nm; flow rate: 1 ml / min; column temperature set to room temperature; injection volume: 10 μl.

[0064] 1 System adaptability investigation

[0065] 1.1 Selection of detection wavelength

[0066] Using the above method, the wavelength scanning function of the diode array detector in the high performance liquid chromatograph was used to perform full wavelength scanning to obtain the ultraviolet absorption spectra of the scopolamine and isofraxidin reference substances ( Fig.16 ). As can be seen from the figure, scopolamine has a maximum absorption at 343nm, and isofraxidin has a maximum absorption at 341nm. Finally, 341nm was determined as the detection wavelength.

[0067] 2 Investigation of the preparation method of test solution

[0068] 2.1 Investigation of extraction solvent

[0069] Take about 1g of the medicinal powder, and make three parallel portions, accurately weigh the weight, place in a round-bottom flask, add 40ml of methanol and ethanol solvents respectively, weigh, cold soak for 1 hour, ultrasonically treat for 30 minutes, cool, make up the lost weight with methanol and ethanol respectively, filter, take the filtrate, inject according to the above chromatic conditions, and analyze. The results are attached. Figure 1-2 and Table 2.

[0070] Table 2 Extraction solvent investigation

[0071]

[0072] The results showed that the extraction efficiency of methanol was higher than that of ethanol, so methanol was determined to be used as the extraction solvent.

[0073] 2.2 Investigation of different extraction methods

[0074] Take about 1g of the medicinal material powder, and weigh three parts in parallel. Place them in round-bottom flasks, add 40ml of methanol respectively, weigh them, and examine three extraction methods: reflux for 30 minutes, ultrasonic for 30 minutes, and cold soak for 1 hour. Filter, take the filtrate, inject it according to the above chromatic conditions, and analyze it. The results are attached. Figure 3-5 and Table 3.

[0075] Table 3 Investigation of different methanol extraction methods

[0076]

[0077] The results showed that the reflux extraction method had the highest efficiency among the three extraction methods, so the reflux extraction method was determined to be used.

[0078] 2.3 Extraction investigation with different methanol concentrations

[0079] Take 1g of medicinal powder, and weigh three portions in parallel. Place in a round-bottom flask, add 40ml of 100% methanol, 80% methanol, and 60% methanol respectively, weigh, soak for 1 hour, reflux for 1 hour, cool, weigh, make up the lost weight with the corresponding solvent, filter, take the filtrate, inject according to the above chromatic conditions, and analyze. The results are attached. Figure 6-8 and Table 4.

[0080] Table 4 Extraction investigation at different methanol concentrations

[0081]

[0082] The results showed that 80% methanol extraction efficiency was optimal, and accordingly the methanol concentration was determined to be 80%.

[0083] 2.4 Extraction time investigation

[0084] Take 1g of medicinal powder, and weigh three portions in parallel, place in round-bottom flasks, add 40ml of 80% methanol, weigh, reflux for 30, 60, 90, 120 minutes, cool, weigh, make up the lost weight with 80% methanol, filter, take the filtrate, inject according to the above chromatic conditions, and analyze. The results are attached. Figure 9-11 and Table 5.

[0085] Table 5 Extraction time investigation

[0086]

[0087] The results showed that the extraction efficiency became stable after reflux extraction for 1.5 hours, and the extraction time was determined to be 1.5 hours.

[0088] 2.5 Investigation of solid-liquid ratio

[0089] Take 1g of medicinal powder, and weigh three portions in parallel. Place in a round-bottom flask, add 10ml, 20ml, 40ml, and 60ml of 80% methanol respectively, weigh, reflux for 1.5h, cool, weigh, make up the lost weight with 80% methanol, filter, take the filtrate, inject according to the above chromatic conditions, and analyze. The results are attached. Figure 12-15 and Table 6.

[0090] Table 6 Solid-liquid ratio investigation results

[0091]

[0092]

[0093] The results showed that the extraction was relatively sufficient when 40 ml of solvent was used, so the amount of solvent was determined to be 40 ml of solvent per 1 gram of medicinal material.

[0094] 2.6 Examination of extraction times

[0095] Take 1g of the medicinal material powder, weigh three times in parallel, place in a round-bottom flask, add 40ml of 80% methanol, weigh, reflux for 1.5h, reflux once, twice, and three times, cool, make up the lost weight, filter, take the filtrate, inject according to the above chromatic conditions, and analyze. The results are shown in Table 7.

[0096] Table 7. Investigation of extraction times

[0097]

[0098] The results showed that one reflux extraction was basically complete, so the number of extractions was determined to be 1.

[0099] According to the above experimental results, the method for preparing the test solution was finally determined as follows: take about 1.0 g of the powder of the product (passed through a No. 3 sieve), accurately weigh it, place it in a stoppered conical flask, add 40 ml of 80% methanol, weigh the weight, heat and reflux for 1.5 hours, cool, weigh the weight again, make up the lost weight with 80% methanol, shake well, filter, and take the filtrate to obtain the product.

[0100] 3 Methodological validation

[0101] 3.1 Specificity test

[0102] The experiment was carried out according to the above method. The results showed that acetonitrile-0.1% formic acid aqueous solution (14:86) was well separated and the blank solvent had no interference. The results are shown in Fig.17 .

[0103] 3.2 Linear investigation

[0104] Accurately pipette 7 portions of the mixed reference solution with different concentrations of dilutions, inject 10 μl of each into the sample, record the chromatographic peak area, and perform linear regression with the peak area (y) as the ordinate and the injection volume (x) as the abscissa. The linear regression equations of scopoletin and isofraginidine are: y=16373x–10533(r 2 =0.9999), y=12583x–12422(r 2 =0.9999); the results are shown in Table 8 and Fig.18 .

[0105] Table 11 Linear relationship investigation results

[0106]

[0107]

[0108] The results showed that there was a good linear relationship between the peak area and isofraxidin in the range of 9.5-950 μg / ml and scopolamine in the range of 4.46-446 μg / ml.

[0109] 3.3 Precision investigation

[0110] Take the same portion of isofragnuine and scopolamine reference solution, inject it six times continuously, record the peak area, calculate its RSD value, and the results are shown in Table 8.

[0111] Table 8 Precision test results

[0112]

[0113] The results showed that after six consecutive injections, the RSD values ​​of isofraxidin and scopolamine were 0.36% and 0.34%, respectively, indicating that the instrument had good precision.

[0114] 3.4 Repeatability test

[0115] Six test solutions were prepared in parallel according to the test solution preparation method, and the samples were injected for measurement and the content was calculated. The RSD value was calculated using the measured content value. The results are shown in Table 9.

[0116] Table 9 Repeatability experiment

[0117]

[0118]

[0119] The results showed that the RSD value of isofraxidin was 0.77%, and the RSD value of scopoletin was 0.85%, indicating that the method had good repeatability.

[0120] 3.5 Stability test

[0121] Take the same batch of Helleborus officinalis medicinal material test solution, and measure it after 0, 2, 4, 8, 12, and 24 hours (injection volume 10 μl), record the peak area, and calculate the RSD value. The results are shown in Table 10.

[0122] Table 10 Solution stability test

[0123]

[0124] The results showed that the RSD value of isofraxidin was 0.46%, and the RSD value of scopolamine was 0.69%, indicating that the solution had good stability within 24 hours.

[0125] 3.6 Recovery test

[0126] Accurately weigh 6 portions of the same batch of medicinal materials with known content, accurately add appropriate amounts of isofraquinone and scopolamine reference substances, prepare the test solution according to the test solution preparation method, inject 10 μl of each, and calculate the sample recovery rate. The results are shown in Table 11.

[0127] Table 11 Determination of sample recovery

[0128]

[0129]

[0130] The results showed that the average recoveries of isofraxidin and scopoletin from the same batch of Helleborus officinalis were 99.30% and 99.54%, respectively. The RSD value of isofraxidin was 0.46%, and that of scopoletin was 0.69%, indicating that the method had a good recovery rate.

[0131] 4. Determination of medicinal material content

[0132] Take 1g of each of 20 batches of Helleborus officinalis medicinal materials, prepare the test solution according to the test solution preparation method, inject the sample under the above chromatographic conditions, measure the peak area, and calculate the content of each component. The results are shown in Table 12.

[0133] Table 12 Content determination results of 20 batches of iron chopstick samples (%)

[0134]

[0135]

[0136] The results showed that the content of isofraxidin ranged from 0.09 to 0.42%, with an average of 0.24%. The average was 0.19% when the average was lowered by 20%. Ten of the 20 batches of samples failed, and the qualified rate was 50%. The content of scopoletin ranged from 0.01 to 0.60%, with an average of 0.25%. The average was 0.20% when the average was lowered by 20%. Ten of the 20 batches of samples failed, and the qualified rate was 50%.

[0137] Among the 20 batches of samples, the contents of isofraquinone and scopolamine varied greatly. As shown in the table, the contents of the two components decreased significantly in samples with a longer storage time (the first 10 batches: samples from 2014 and 2015), which may be related to the storage time and storage conditions of the samples.

[0138] Considering factors such as ensuring the quality of medicinal materials and encouraging the improvement of storage conditions of medicinal materials, it is determined that isofraxidin (C 11 H 10 O5) is limited to not less than 0.12% (the qualified rate of 20 batches of samples is 65%); Scopoletin (C 10 H8O4) shall not be less than 0.20% (the qualified rate of 20 batches of samples is 50%).

Claims

1. A quality detection method for Helleborus officinalis medicinal material, characterized in that: The invention comprises the determination of the content of isofraxidin and scopolamine in the medicinal material of Helleborus chinensis; the content determination adopts ultra-high performance liquid chromatography tandem time-of-flight mass spectrometry experimental technology to determine the content of isofraxidin and scopolamine in the medicinal material of Helleborus chinensis; the method for determining the content of isofraxidin and scopolamine in the medicinal material of Helleborus chinensis is determined according to high performance liquid chromatography, and comprises the following steps: (1) Determine the chromatographic conditions Chromatographic column: Cosmosil C 18 Column; mobile phase: acetonitrile: 0.1%, formic acid water = 14:86; detection wavelength: 341nm; flow rate: 1ml / min; column temperature is room temperature; injection volume: 10μl; (2) Preparation of reference solution: Take appropriate amount of scopoletin and isofraxidin reference substances, weigh accurately, and add methanol to prepare a mixed solution containing 0.066 mg of scopoletin and 0.065 mg of isofraxidin per 1 ml to obtain the reference solution; (3) Preparation of test solution: Take about 1.0 g of the powder of the product, weigh it accurately, place it in a stoppered conical flask, add 40 ml of 80% methanol accurately, weigh it, heat and reflux to extract for 1.5 hours, cool, weigh it again, make up the lost weight with 80% methanol, shake well, filter, and take the filtrate to obtain the test solution; (4) Determination method: Accurately pipette 10 μl of the reference solution and the test solution respectively, inject into the liquid chromatograph, and determine.

2. The quality detection method of Helleborus officinalis medicinal material according to claim 1, characterized in that: This product contains isoflurane C based on dry product. 11 H 10 O5 shall not be less than 0.12%, Scopoletin C 10 H8O4 shall not be less than 0.20%.

Images