Application of an integrated quality safety assessment method in Huangjingzanyu capsules
The heavy metals in Polygonatum Pangyu Capsules were detected through microwave digestion and inductively coupled plasma mass spectrometry. Combined with risk assessment, the problem of heavy metal pollution in traditional Chinese medicinal materials was solved, ensuring the safety of drugs and drug use safety, and avoiding the potential of heavy metals on the human body. harm.
Patent Information
- Application Number
- CN202110325068.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2021-03-26
- Publication Date
- 2024-12-10
- Estimated Expiration
- 2041-03-26
AI Technical Summary
Heavy metal contamination in traditional Chinese medicinal materials affects the safety of drugs, especially in long-term use, which may lead to human health risks. It is necessary to accurately detect and limit the content of heavy metal elements to ensure the safety of medication.
The microwave digestion method is used to digest Polygonatum Pillaria capsule powder, and the heavy metal content is detected by inductively coupled plasma mass spectrometry, and the risk assessment method is combined to evaluate its potential harm to human health.
It has achieved accurate detection and risk assessment of heavy metal elements in Polygonatum Pangyu Capsules, ensured the safety of clinical medication, avoided the potential harm of heavy metal accumulation to the body, and provided a solid foundation for quality and safety evaluation.
Smart Images
Figure FDA0004967907160000011 
Figure FDA0004967907160000021
Abstract
Description
Technical Field
[0001] The invention belongs to the field of drug safety evaluation, and particularly relates to application of an integrated quality safety evaluation method in Huangjing Zanyu Capsules. Background Art
[0002] Huangjing Zanyu Capsule is composed of 20 Chinese medicinal materials including Huangjing (made with wine), Heshouwu (made with wine), Schisandra chinensis, Lycium barbarum, Placenta, Epimedium, Cuscuta chinensis, Rehmannia glutinosa, Cistanche deserticola, Dipsacus asper, Codonopsis pilosula, Salvia miltiorrhiza, Angelica sinensis, Taraxacum officinale, Patrinia scabra, Cnidium monnieri, Beehive (fried), Leech, Oyster, and Plantago asiatica (fried with salt), and is a compound Chinese medicinal preparation made from some raw powder and some extracted materials. It has the effects of tonifying the kidney and replenishing essence, clearing away heat and removing dampness, and is used for male infertility caused by asthenospermia and oligospermia of kidney deficiency and essence deficiency with damp-heat type. The prescription contains multiple root or rhizome medicinal materials such as Angelica sinensis, Salvia miltiorrhiza, Huangjing, Heshouwu, Dipsacus asper, Codonopsis pilosula, and Rehmannia glutinosa, which are easy to enrich heavy metals and harmful elements in the soil and water during their growth, thus introducing quality risks into the preparation.
[0003] Excessive heavy metal content in traditional Chinese medicine has become a focal issue for drug safety and human health at home and abroad, and is also a bottleneck restricting the modernization and internationalization of traditional Chinese medicine. In the context of protecting human health and drug safety, relevant state ministries and commissions have formulated and improved corresponding quality control standards for residual harmful heavy metals in medicinal plants and preparations. Since the use cycle of traditional Chinese medicine is generally longer than that of chemical medicine, heavy metals are prone to accumulation in the body, so the prevention and control of heavy metal pollution in traditional Chinese medicine should be paid special attention. In recent years, the problem of excessive heavy metal elements in traditional Chinese medicine has attracted much attention. Heavy metal elements and harmful elements in traditional Chinese medicine generally include lead (Pb), mercury (Hg), cadmium (Cd), arsenic (As), copper (Cu), etc., which can cause harm to the nervous system, digestive system, hematopoietic system, metabolism, etc. after acting on the human body.
[0004] Excessive heavy metal content has become one of the main issues affecting the development of traditional Chinese medicine. Accurate detection and limitation of the content of heavy metal elements is the key to ensuring the safety and effectiveness of people's use of medicine. In 2015, the state successively issued the international standard "Limits of Heavy Metals in Traditional Chinese Medicine-Chinese Medicinal Materials" formulated by the China Academy of Chinese Medical Sciences and the "Notice of the State Administration of Traditional Chinese Medicine on Further Strengthening the Quality Management of Chinese Medicine Pieces Prescriptions and Strengthening Rational Use" issued by the State Administration of Traditional Chinese Medicine. It can be seen that the state attaches great importance to the content of heavy metal elements in Chinese medicinal materials. Based on the provisions of the "Green Industry Standards for Import and Export of Medicinal Plants and Preparations", Zhao Hualian et al. counted the overall situation of heavy metal pollution in Chinese medicinal materials in my country and found that Pb, Cd, Hg, As, and Cu all have varying degrees of pollution.
[0005] Therefore, the present invention provides an application of an integrated quality safety assessment method in Huangjing Zanyu Capsule to improve the safety of its clinical use. Summary of the invention
[0006] Based on the above situation, the purpose of the present invention is to provide an integrated method for quality and safety assessment for application in Huangjing Zanyu Capsules.
[0007] The object of the present invention is achieved through the following technical scheme: an application of an integrated method for quality and safety assessment in Huangjing Zanyu Capsules, the main steps of which include: step 1: subjecting Huangjing Zanyu Capsules powder to microwave digestion; step 2: subjecting the Huangjing Zanyu Capsules digestion solution to heavy metal analysis; step 3: conducting a risk assessment on the analysis results to evaluate the hazard risk of heavy metal content in Huangjing Zanyu Capsules.
[0008] In one embodiment of the present invention, the microwave digestion method is characterized in that the specific steps are as follows: Step 1: Place the Huangjing Zanyu Capsule powder in a digestion tank and add a digestion solution into the digestion tank; Step 2: Place the digestion tank open in a fume hood for pre-digestion; Step 3: Seal the digested solution and place it in a microwave digestion instrument; Step 4: After the digestion is completed, place the digestion tank on a hot plate to drive out the acid, evaporate the digestion solution to 1-2 mL, transfer it to a volumetric flask, rinse the digestion tank and the lid with ultrapure water for 3-5 times, and similarly transfer the washing solution to a volumetric flask.
[0009] In one embodiment of the present invention, the Huangjingzanyu capsules are pre-processed by removing the capsule shell and crushing before digestion to make them into powder.
[0010] In one embodiment of the present invention, the Huangjing Zanyu Capsule powder is placed in a digestion tank for microwave digestion, wherein the digestion solution used in the microwave digestion is a mixed solution of nitric acid and hydrogen peroxide.
[0011] In one embodiment of the present invention, the mixed solution of nitric acid and hydrogen peroxide is characterized in that the volume ratio of nitric acid to hydrogen peroxide is 5:1.
[0012] In one embodiment of the present invention, the Huangjing Zanyu Capsule powder is placed in a digestion tank for microwave digestion, wherein the mass ratio of the Huangjing Zanyu Capsule powder to the digestion solution is 1:40.
[0013] In one embodiment of the present invention, the microwave digestion method, wherein the temperature raising program of the microwave digestion instrument is: first raise the temperature to 100°C and keep it warm for 5 minutes; then raise the temperature to 130°C and keep it warm for 5 minutes; finally raise the temperature to 160°C and keep it warm for 20 minutes.
[0014] In one embodiment of the present invention, the digestion tank is placed on a hot plate to drive out the acid, characterized in that the heating temperature of the hot plate is 120°C.
[0015] In one embodiment of the present invention, the digested solution is subjected to heavy metal analysis, wherein the method adopted comprises inductively coupled plasma mass spectrometry.
[0016] In one embodiment of the present invention, the risk assessment of the analysis results is characterized by comprising the following steps:
[0017] Step 1: Hazard identification, generally achieved through human epidemiological information or animal toxicology testing.
[0018] Step 2: Exposure assessment, which is a qualitative or quantitative evaluation of the intake of hazardous substances, generally expressed as daily / weekly / monthly heavy metal intake per kilogram of body weight.
[0019] Step 3: Hazard characterization, generally achieved through animal toxicity evaluation information or human epidemiological information.
[0020] Step 4: Risk characterization, that is, based on the results of the three steps of hazard identification, hazard characterization and exposure assessment, the possibility and severity of known or potential adverse health effects for a specific population are assessed.
[0021] The present invention applies the inductively coupled plasma mass spectrometry heavy metal content detection method to Huangjing Zanyu Capsule for the first time, and integrates the risk assessment method to evaluate the analysis results, analyze the risk of harm to human health, and evaluate the quality and safety of the large compound preparation Huangjing Zanyu Capsule to avoid harm to the body caused by heavy metal accumulation in patients taking the medicine, providing a solid foundation for the clinical drug safety of Huangjing Zanyu Capsule. DETAILED DESCRIPTION
[0022] The following examples are provided for a better understanding of the present invention, but are not intended to limit the best mode of implementation, nor to limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior arts shall fall within the protection scope of the present invention.
[0023] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be obtained commercially.
[0024] Example: Application of an integrated quality safety assessment method in Huangjing Zanyu Capsules
[0025] 1. Microwave digestion of Huangjingzanyu capsules
[0026] Take samples according to the method in Table 1, add digestion solution, pre-digest for 60 minutes, and digest according to the program in Table 2. After digestion, cool to room temperature, drive out the acid to 1-2mL, transfer to a tube, wash the microwave digestion tank several times with ultrapure water, combine the washing solution with the test solution, and make up to 10mL.
[0027] The digestion effect is judged by whether there is residue in the digestion solution after digestion. After comparing the digestion solutions, it was found that no residue was visible to the naked eye in digestion tanks No. 1 and No. 2, while there was a small amount of residue that could just be observed by the naked eye in digestion tanks No. 5 and No. 6. Therefore, it is speculated that adding 2mL is beneficial to the digestion of Huangjing Zanyu Capsule powder. Subsequently, the optimal digestion conditions will be indirectly determined by comparing the contents of different heavy metals.
[0028] Table 1 Experimental conditions
[0029] serial number Mass / g <![CDATA[Volume of HNO3 / mL]]> <![CDATA[Volume of H2O2 / mL]]> 1 0.3 10 2 2 0.3 10 2 3 0 10 0 4 0 10 0 5 0.3 10 0 6 0.3 10 0
[0030] Table 2 Digestion conditions
[0031] step Power / W Heating time / min Temperature / ℃ Duration / min 1 1600 5 100 5 2 1600 5 130 5 3 1600 5 160 20
[0032] 2. Determination of heavy metal content in Huangjingzanyu capsules
[0033] 2.1 Optimization of instrument parameters
[0034] The heavy metal content of the pretreated Huangjing Zanyu capsule digestion solution was determined by inductively coupled plasma mass spectrometry (ICP-MS) technology using the standard curve method. Before the experiment, the instrument was tuned intelligently (SmartTune) to automatically adjust the instrument to the optimal state. The instrument conditions were as follows: plasma RF power 1600W, sampling depth 2.0mm, plasma gas flow rate 15.0L·min -1 , atomizing gas flow rate 1.0L·min -1 , auxiliary gas flow rate 1.2L·min -1 The software parameter settings are shown in Table 4.
[0035] Table 4 Software parameter settings
[0036] element Mass number (amu) Measurement Mode Responsive Mode He flow R R As 75 Peak Jump KED 3.5 0 0.25 Cd 111 Peak Jump KED 3.5 0 0.25
[0037] Cu 63 Peak Jump KED 3.5 0 0.25 Pb 208 Peak Jump Standard 0 0.25 Hg 202 Peak Jump Standard 0 0.25
[0038] 2.2 Preparation of test solution
[0039] Take Huangjing Zanyu Capsule and dry it at 60℃ for 2 hours, crush it into coarse powder, take 0.3g, weigh it accurately, put it in a pressure and high temperature microwave digestion tank, add 10mL nitric acid and 2mL hydrogen peroxide (if the reaction is violent, leave it until the reaction stops). Seal it and digest it according to the corresponding requirements of each microwave digestion instrument and a certain digestion procedure. After the digestion is complete, cool the digestion solution to below 60℃, take out the digestion tank, let it cool, drive the acid to 1-2mL, transfer the digestion solution to a 10mL volumetric bottle, wash the digestion tank and the lid with a small amount of water 3 times, combine the washing solution in the volumetric bottle, dilute it with water to the scale, shake it well, and you have it.
[0040] 2.3 Preparation of standard solution
[0041] Standard solutions of lead, chromium, arsenic and copper with concentrations of 1, 2.5, 10 and 20 μg / L, respectively, and standard solutions of mercury with concentrations of 0.2 and 1.0 μg / L, respectively, were prepared. The rest of the preparation method was the same as that of the test sample to obtain a calibration standard curve; the content of lead, chromium, cadmium, copper and mercury in the test solution was detected by inductively coupled plasma mass spectrometry (ICP-MS).
[0042] 2.4 Content determination
[0043] In general, the average contents of the five heavy metals in Huangjing Zanyu Capsules are Cu, Pb, As, Hg, and Cd from high to low, as shown in Table 6. According to the heavy metal limit regulations in the International Standard for Heavy Metal Limits in Chinese Herbal Medicines (ISO18664:2015) and the Green Industry Standard for Import and Export of Medicinal Plants and Preparations: The total amount of heavy metals is ≤20 mg kg -1 、Cu≤20mg·kg -1 、As≤2mgkg -1 、Cd≤0.3mg·kg -1 、Pb≤5mg·kg -1 、Hg≤0.2mg·kg -1 It can be seen that the contents of five heavy metal elements in Huangjing Zanyu Capsule did not exceed the limit standards.
[0044] At the same time, it can be seen from the experimental results that the heavy metal detection amounts in the groups with hydrogen peroxide added during digestion (i.e., groups 1 and 2) are mostly higher than those in the groups without hydrogen peroxide added (i.e., groups 5 and 6). This shows that adding hydrogen peroxide during digestion is beneficial to a more complete digestion of the Huangjing Zanyu Capsule powder.
[0045] Table 6 Content determination results
[0046] serial number As (μg / kg) Cd(μg / kg) Cu(μg / kg) Pb(μg / kg) Hg(μg / kg) 1 0.77 0.065 5.0 1.0 0.094 2 0.74 0.066 4.9 1.0 0.059
[0047] 3 0.13 0.0010 1.7 0.059 0.017 4 0.10 0 1.4 0.051 0.010 5 0.66 0.059 4.6 0.43 0.035 6 0.71 0.068 5.0 0.43 0.031
[0048] 3. Risk Assessment
[0049] Risk assessment generally includes four basic steps: hazard identification, exposure assessment, hazard characterization and risk characterization. In actual work, hazard identification and hazard characterization are often achieved through animal toxicity evaluation information or human epidemiological information or through animal toxicity tests. By determining the quantitative relationship between hazardous substances and adverse reactions in the human body, that is, determining the dose-response mathematical model, the no observed adverse effect level (NOAEL) of harmful residues can be determined. The health guidance value is calculated through NOAEL, that is, the tolerable weekly intake per kilogram of body weight (PTWI), the tolerable monthly intake per kilogram of body weight (PTMI) or the acceptable daily intake (ADI). Exposure assessment refers to the qualitative or quantitative evaluation of the intake generated by possible ingestion or other various routes. The National Sanitation Foundation (NSF) of the United States believes that the amount of heavy metals ingested daily from dietary supplements accounts for 10% of the total daily exposure (the total amount of heavy metals ingested from various routes such as food and drinking water) and will not cause health risks. Risk characterization is an assessment of the likelihood and severity of known or potential adverse health effects on a specific population based on the results of three steps: hazard identification, hazard characterization, and exposure assessment. It can usually be achieved through the ratio of exposure to ADI, PTDI, PTWI, etc.
[0050] According to the above general steps, the risk assessment of heavy metals and harmful elements in Huangjing Zanyu Capsules was carried out. Arsenic was used as an example to illustrate the calculation process of the hazard quotient (HQ) of each element: The instructions for Huangjing Zanyu Capsules stipulate that the dosage of this product is "4 capsules at a time, 3 times a day", with a specification of 0.31g / capsule. -1 , then the daily dosage F = 4 × 3 × 0.31 = 3.72 g. From Table 6, we can see that the residual arsenic content C is 15.60 mg kg -1 According to formula (1), the daily intake of arsenic per kilogram of body weight Exp = F × C / BW = 3.72 g × 7.75 mg kg -1 / 60kg=0.48μg·kg -1 The acceptable daily intake (ADI) of arsenic per kilogram of body weight is 2.1 μg kg -1 According to formula (2), HQ = Exp / ADI = 0.48 μg·kg -1 / 2.1μg·kg -1 =0.23. Similarly, the HQ of other elements and their sum HI are calculated, see Table 7.
[0051] The risk order of the five elements is lead>arsenic>copper>mercury>cadmium. The total hazard index of heavy metals and harmful elements in Huangjing Zanyu Capsule is less than 1, indicating that this variety does not have non-carcinogenic risks. The above results show that the quality control of Huangjing Zanyu Capsule is good, the production process is standardized, and the heavy metal content is far below the limit. Even patients who take it for a long time do not have health threats.
[0052] Table 7 Exp, HQ, HI and contribution rate of 5 elements in HI
[0053] element <![CDATA[Exp / μg·kg -1 ]]> HQ Contribution rate in HI (0.61) / % <![CDATA[ 75 As]]> 0.48 0.23 37.7 <![CDATA[ 111 Cd]]> 0.052 0.05 8.2 <![CDATA[ 63 With]]> 3.06 0.06 9.8 <![CDATA[ 202 Hg]]> 0.058 0.10 16.4 <![CDATA[ 208 pb]]> 0.63 0.17 27.9
[0054] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.
Claims
1. Application of an integrated quality safety assessment method in Huangjing Zanyu Capsule, characterized in that: The Huangjing Zanyu Capsule formula includes seven kinds of root and rhizome medicinal materials, including Polygonum multiflorum, Salvia miltiorrhiza, Angelica sinensis, Polygonatum sibiricum, Dipsacus asper, Codonopsis pilosula, and Rehmannia glutinosa, which have a high risk of heavy metal pollution. The method includes the following steps: Step 1: performing microwave digestion on the pre-treated Huangjingzanyu capsules; Step 2: Analyze the heavy metal content of the digestion solution of Huangjingzanyu capsule; Step 3: Conduct risk assessment on the analysis results to evaluate the hazard risk of heavy metal content in Huangjing Zanyu Capsules. The microwave digestion method described in step 1 comprises the following specific steps: Step (1): placing the Huangjing Zanyu Capsule powder in a digestion tank, and adding a digestion solution into the digestion tank, wherein the digestion solution used is a mixed solution of nitric acid and hydrogen peroxide, and the volume ratio of nitric acid to hydrogen peroxide is 5:1; Step (2): Place the digestion tank open in a fume hood for pre-digestion; Step (3): The digested solution is sealed and placed in a microwave digestion instrument. The heating program of the microwave digestion instrument is as follows: first, the temperature is raised to 100° C. and kept warm for 5 minutes; then the temperature is raised to 130° C. and kept warm for 5 minutes; finally, the temperature is raised to 160° C. and kept warm for 20 minutes; Step (4): After digestion, place the digestion tank on a hot plate to remove the acid, evaporate the digestion solution to 1-2 mL, transfer it to a volumetric flask, rinse the digestion tank and lid with ultrapure water 3-5 times, and transfer the washing solution to the volumetric flask. The method described in step 2 is an inductively coupled plasma mass spectrometry method for simultaneously determining multiple elements. The standard curve method is used to determine the heavy metal content of the pretreated Huangjing Zanyu Capsule digestion solution. Before the experiment begins, the instrument is intelligently tuned to automatically adjust the instrument to the optimal state. The instrument conditions are as follows: plasma RF power 1600W, sampling depth 2.0mm, plasma gas flow rate 15.0L·min -1 , atomizing gas flow rate 1.0L·min -1 , auxiliary gas flow rate 1.2L·min -1 , the software parameters are set as follows: The specific steps of step 3 are as follows: Step (1): Hazard identification, which is achieved through human epidemiological information or animal toxicology tests by determining the quantitative relationship between hazardous substances and adverse reactions in humans, that is, determining a dose-response mathematical model to determine the no observed adverse effect level (NOAEL) of hazardous residues, and calculating the health guidance value through NOAEL, that is, the tolerable weekly intake per kilogram of body weight (PTWI), the tolerable monthly intake per kilogram of body weight (PTMI) or the acceptable daily intake (ADI); Step (2): Exposure assessment, i.e. qualitative or quantitative evaluation of the intake of hazardous substances, expressed as daily / weekly / monthly intake of heavy metals per kilogram of body weight; Step (3): Hazard characterization, achieved through animal toxicity evaluation information or human epidemiological information; Step (4): Risk characterization, that is, based on the results of the three steps of hazard identification, hazard characterization and exposure assessment, the possibility and severity of known or potential adverse health effects for a specific population are evaluated through the ratio of exposure to ADI, PTDI and PTWI.
2. Application of the integrated quality safety assessment method according to claim 1 in Huangjingzanyu Capsule, characterized in that: In step 1, the Huangjingzanyu capsules are shelled and crushed before digestion to make them into powder.
3. The method of placing Huangjingzanyu capsule powder in a digestion tank for microwave digestion according to claim 1, characterized in that: In step 1, the mass ratio of Huangjing Zanyu Capsule powder to digestion solution is 1:40.