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Development of the Chinese Pharmacopoeia 2020 Edition General Chapters: A Review
Abstract and Figures
The 2020 edition of the Chinese Pharmacopoeia was reviewed and approved by the National Medical Products Administration and the National Health Commission of the People's Republic of China in July 2020. The current edition was officially implemented on December 30, 2020. The general chapters of the Chinese Pharmacopoeia discuss the general testing methods and guidelines, which are the common requirements and basis for the implementation of drug standards in the Chinese Pharmacopoeia. Owing to adherence to the principles of scientificity, versatility, operability, and sustainable development, there was an improvement in the general chapters of the 2020 edition compared to those of the previous editions. Further, the application of advanced and mature analytical techniques has expanded, the development of testing methods for exogenous pollutants in traditional Chinese medicines has been strengthened, and technical requirements are now better harmonized with international standards. The updated edition provides technical and methodological support to ensure safety, effectiveness, and control of pharmaceuticals in China and will play an important and active role in encouraging the application of advanced technologies, improving the quality control of medicines, and strengthening the means of drug regulation in China. This review provides a comprehensive introduction about the main features of and changes to the general chapters in the 2020 Chinese Pharmacopoeia and aims to provide references for its correct understanding and accurate implementation.
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Review paper
Development of the general chapters of the Chinese Pharmacopoeia
2020 edition: A review
Xinyi Xu, Huayu Xu, Yue Shang, Ran Zhu, Xiaoxu Hong, Zonghua Song, Zhaopeng Yang
*
Chinese Pharmacopoeia Commission, Beijing, 100061, China
article info
Article history:
Received 27 December 2020
Received in revised form
5 May 2021
Accepted 9 May 2021
Available online 20 May 2021
Keywords:
Chinese pharmacopoeia
2020 edition
General chapter
Development
Review
abstract
The Chinese Pharmacopoeia 2020 edition was reviewed and approved by the National Medical Products
Administration and the National Health Commission of the People's Republic of China in July 2020. The
current edition was offi cially implemented on December 30, 2020. The general chapters of the Chinese
Pharmacopoeia discuss the general testing methods and guidelines, which are the common re-
quirements and basis for the implementation of drug standards in the Chinese Pharmacopoeia. Owing to
adherence to the principles of scientifi city, versatility, operability, and sustainable development, there is
an improvement in the general chapters of the 2020 edition over those of the previous editions. Further,
the application of advanced and mature analytical techniques has expanded, the development of testing
methods for exogenous pollutants in traditional Chinese medicines has been strengthened, and technical
requirements are now better harmonized with international standards. The updated edition provides
technical and methodological support to ensure safety, effectiveness, and control of pharmaceuticals in
China and will play an important and active role in encouraging the application of advanced technolo-
gies, improving the quality control of medicines, and strengthening the means of drug regulation in
China. This review provides a comprehensive introduction of the main features of and changes to the
general chapters in the Chinese Pharmacopoeia 2020 edition and aims to provide reference for its correct
understanding and accurate implementation.
©2021 Xi'an Jiaotong University. Production and hosting by Elsevier B.V. This is an open access article
under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
1. Introduction
The Chinese Pharmacopoeia 2020 edition was reviewed and
approved by the National Medical Products Administration (NMPA)
and the National Health Commission of the People's Republic of
China in July 2020. This edition was offi cially implemented on
December 30, 2020. The Chinese Pharmacopoeia is a statutory
technical specifi cation that must be implemented for drug devel-
opment, production, use, and regulation in China. The general
chapters in the Chinese Pharmacopoeia are the basis for the accu-
rate implementation of the Chinese Pharmacopoeia. The 2020
edition contains 360 general chapters, including 23 new and 83
revised chapters. This updated edition refl ects not only the current
level of technology used in the pharmaceutical industry in China
but also the technologies used for international drug quality
control.
2. Application of advanced and mature analytical techniques
has been expanded
Method 0451, " X-ray fl uorescence spectroscopy" , was added to
guide the application of X-ray fl uorescence spectroscopy in the
qualitative and quantitative analyses of elemental impurities [1e9].
Oscillating transducer density meter application and instrumen-
tation were added in method 0601, " Determination of Relative
Density" [10 ,11 ]. In method 0713, " Tests of Fat and Fatty Oil" , the
melting range, saponifi cation value, and iodine value were revised,
and information concerning unsaponifi able matter, fatty acid
composition, alkaline impurities, anisidine value, sterols, and trans
fatty acids was added [12e 16 ]. Methods 1001, " Polymerase Chain
Reactions" ;1021,"Identifi cation of Bacterial DNA Sequences "; and
9108, " DNA Sequencing" , were added. These methods are used to
ensure the accurate identifi cation and clinical safety of drugs
[17e22 ]. In vitro methods, which involve the use of an instrument
to determine endpoints, have replaced in vivo biological methods,
which is in line with the goal of reducing, replacing, and refining
laboratory animal use. The anti-factor IIa and anti-factor Xa assays
Peer review under responsibility of Xi'an Jiaotong University.
*Corresponding author.
E-mail address: yangzhaopeng@chp.org.cn (Z. Yang).
Contents lists available at ScienceDirect
Journal of Pharmaceutical Analysis
journal homepage: www.elsevier.com/locate/jpa
https://doi.org/10.1016/j.jpha.2021.05.001
2095-1779/© 2021 Xi'an Jiaotong University. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.
org/licenses/by-nc-nd/4.0/).
Journal of Pharmaceutical Analysis 11 (2021) 398e404
were added to method 1208, " Biological Assay of Heparin" [ 23e 27 ].
The heparin-binding capacity assay was added to method 1213,
"Biological Assay of Protamine Sulfate "[28e32 ]. New sterilization
methods (vapor-phase sterilization and liquid-phase sterilization)
were added to method 1421, " Methods of Sterilization" , to guide the
sterilization/fi ltration-based production of drugs to ensure that the
sterility level meets the requirements [33e36 ]. The monocyte
activation test was added in guideline 9301, " Application of Safety
Tests for Injection" [37e 42]. Based on research detailing the tech-
nical requirements for the verifi cation and transfer of analytical
methods from the United States Pharmacopoeia (USP) and the
American Association of Analytical Chemists [43e 49 ] to the Chi-
nese Pharmacopoeia, guidelines 9099, "Verifi cation of Compendial
Procedures" , and 9100, " Transfer of Analytical Procedures" ,were
added.
The applicability requirements of the analytical methods were
enhanced. In method 1105, " Microbiological Examination of Non-
sterile Products: Microbial Enumeration Tests" , an improved
method for the preparation of aerosol test samples was included,
and information regarding the quantities of small-dose, low-con-
tent, and small-batch samples to be tested was added. In method
1107, " Microbiological Acceptance Criteria of Nonsterile Pharma-
ceutical Products" , the microbiological acceptance criteria for
semisolid preparations were modified to ensure strict control, as is
required for liquid preparations. The defi nition and scope of anti-
microbial preservatives in method 1121, " Antimicrobial Effective-
ness Testing" , have been revised to enhance accuracy. The recovery
rates of the suitability test for the medium and test microorganisms
used in the operational suitability test were revised to ensure
consistency with method 1105 [50 ]. Other newly added and revised
general testing methods are detailed in Table 1.
3. Development of testing methods for exogenous pollutants
in traditional Chinese medicines (TCMs) has been
strengthened
Methods for determining exogenous pollutants in TCMs have
been improved in the Chinese Pharmacopoeia 2020 edition.
Qualitative screening and quantitative analytical methods,
including gas chromatography-tandem mass spectrometry (GC-
MS/MS) and high-performance liquid chromatography-tandem
mass spectrometry (HPLC-MS/MS), were added to method 2341,
"Determination of Pesticide Residues". Qualitative screening
methods are used for the rapid qualitative testing, risk monitoring,
and early warning testing of pesticides. Pesticides with limited
requirements can be directly determined using a quantitative
analytical method. In the 2020 edition, the number of pesticides
tested has increased to 592. Eighty-eight pesticides were identified
using GC-MS/MS and 523 pesticides were determined using HPLC-
MS/MS. For pesticides that can be determined with GC-MS/MS and
HPLC-MS/MS, the preferred method is provided, and the maximum
possible number of characteristic ions is recommended [51e56 ].
Accurate and low-detection-limit methods, including HPLC-MS/
MS analysis of afl atoxin and patulin; HPLC and HPLC-MS/MS ana-
lyses of ochratoxin A, zearalenone, and vomitoxin; and HPLC-MS/
MS analysis of multiple mycotoxins, were added to method 2351,
"Determination of Mycotoxins ". Considering that the above-
mentioned methods require complex sample pretreatment, spe-
cifi c instruments, and specialized personnel training, they will be
subject to limitations for being used in quality control of TCMs. A
fast, sensitive, simple, and low-cost afl atoxin ELISA method was
added as a new technique for the quality control of TCM in China
(Table 2 )[ 57e63].
As the development of acceptable microbiological criteria for
TCM decoction pieces was a breakthrough, strategies and methods
for specifi c microbial contamination control in TCMs with different
uses were introduced. Compared with the medicines produced
according to good manufacturing practices, TCM decoction pieces
contain microorganisms in high abundance, with a wider variety of
species and a more uneven distribution. Further, the microbial
analysis requirements are unique to each type of medicinal mate-
rial. Therefore, method 1108, " Microbiological Examination of
Traditional Chinese Medicine Decoction Pieces" , was added. The
enumerated microbial parameters include total aerobic microbial
counts, total combined yeast/mold counts, and the number of heat-
resistant bacteria; the parameters for specifi ed microorganisms
Table 1
Additions and revisions in the Chinese Pharmacopoeia 2020 edition general testing methods.
General chapter Additions and revisions
0421, Raman Spectroscopy Transmittance, tip-enhanced Raman spectroscopy, and imaging techniques were introduced, and their applications in
the fi elds of physics, chemistry, process control, and other analyses was expanded.
0512, High-Performance Liquid Chromatography The latest HPLC developments and application progress were fully detailed. Information about multidimensional HPLC,
charged aerosol detection, adjusted chromatographic conditions, and common qualitative analysis methods was added.
0661, Thermal Analysis A thermogravimetry-mass spectrometry method was added to realize the qualitative and quantitative analyses of
crystallization solvents (aqueous) or other volatile components in a test sample
0981, Crystallinity Differential scanning calorimetry was added for crystallinity tests of the sharp endothermic peak of crystalline
materials or the dispersion (or no endothermic peak) characteristics of amorphous materials. This method can also be
used to identify the crystalline form when there is a difference in the endothermic peak position of the solid state of
different crystal forms of the same compound.
0991, Determination of Specifi c Surface Area;
0992, Determination of the Density of Solids
The basic defi nitions and terminology of specifi c surface area and solid density are given. Information about
instruments and measuring methods is provided.
1143, Test for Bacterial Endotoxins;
9251, Guideline for the Application of the
Bacterial Endotoxin test
Traceability with international standards and a description of false-positive results and processing methods were added
to avoid misinterpretation due to
b
-glucans. The gel-clot method was revised, and the requirement to initially fi ll the
amoebocyte lysate, followed by the addition of endotoxin, was removed. To standardize the design, procedure, and
limit setting of interference experiments, to ensure even quality, and to address a lack of amoebocyte lysate,
information about the contents of the bacterial endotoxin for limit setting, choice of methods, and pretreatment
methods for test samples were added. The recombinant factor C assay, which is suitable for testing samples containing
b
-glucans, factor B, and prothrombin, was introduced to address the shortage of amoebocyte lysate resources.
1146, Test for Histamines;
9301, Guideline for the Application of Safety Tests
for Injections
The preparation method for the test histamine solution, the method suitability test, and the determination of the
minimum valid concentration test were added.
9015, Guideline for Studies and Quality Control of
Drug Polymorphisms
Solid nuclear magnetic resonance spectroscopy method was added. The differences in the chemical environment of the
same atomic nucleus of different crystal forms of a test sample causes differences in chemical shifts, coupling constants,
and relative intensities during identifi cation of the crystalline states.
X. Xu, H. Xu, Y. Shang et al. Journal of Pharmaceutical Analysis 11 (2021) 398e404
399
include the number of bile-tolerant gram-negative bacteria,
Escherichia coli, and Salmonella. The quantity of the product to be
tested, the preparation method for the test solution, and the suit-
ability test of the counting method are specifi ed; and the uncer-
tainty in the interpretation of the results can be greater for TCMs
than that for other products.
In method 2322, " Determination of Mercury and Arsenic
Speciation and Valence States" , the method of test solution prep-
aration was improved to address the diffi culties in determining the
valence states of arsenic and mercury in marine- and animal-
derived TCMs. Notably, information regarding the preparation of
the test solution, determination of the sample amount, and prin-
ciple of the method application, was added.
4. Technical requirements are better harmonized with the
International Council for Harmonization (ICH) of technical
requirements for pharmaceuticals for human use guidelines
In 2017, the NMPA joined the ICH. In the process of compiling
the Chinese Pharmacopoeia 2020 edition, the implementation of
international standards was further strengthened (Table 3)
[64e67 ]. Considering the current status of drug production and
Table 2
Comparison of immunological and chemical methods for the determination of afl atoxins.
Item Pretreatment Sensitivity Type Percent recovery (%) Equipment Speed Cost
(RMB/
test)
Personnel Refs.
Hordei Fructus
Germinatus
Ziziphi
Spinosae
Semen
Persicae
Semen
Coicis
Semen
ELISA Direct dilution or
extraction, 20e60 min
ng/mL AFB
1
82.8e 95.9 74.7 e 88.5 94.1 e 101.9 84.0 e 89.1 Fluorophotometer
(10,000e 30,000 RMB)
60
min/
test
30e 50 Ordinary personnel
can operate
[58]
AFTs 98.4e 110.4 87.0e 98.6 104.0e 112.0 86.8e103.3
HPLC Immunoaffinity
column, 4e 8h
ng/mL AFB
1
61.3e 70.9 57.0 e 61.8 68.1 e 78.5 63.9 e 69.7 HPLC (< 100,000 RMB) 3 h/
test
200
e300
Requires trained
personnel to operate
[58]
AFTs 62.5e 78.5 58.1e 67.8 69.2e 84.0 65.1e74.5
AFB1: afl atoxin B1; AFTs: total aflatoxin.
Table 3
Implementation status of the ICH Q4B in the Chinese Pharmacopoeia 2020 edition.
ICH No./Chinese
Pharmacopoeia
No.
Testing method Implementation
status
Main differences Refs.
Annex 1/0841 Residue on ignition/sulfated ash In the process of
implementation
Sulfuric acid addition amount, ignition temperature, and
conditions for the end of the experiment
[64e66]
Annex 2/0102,
0942
Test for extractable volume of parenteral preparations In the process of
implementation
Sampling method, method details, and result interpretation [64e66]
Annex 3/0903 Test for particulate contamination: subvisible particles In the process of
implementation
Instrument calibration for the light obscuration particle count
test, requirements for testing environmental water samples,
sampling method, and the evaluation of injections with a labeled
volume of 100 mL
[64e66]
Annex 4A/1105 Microbiological examination of nonsterile products:
microbial enumeration tests
In the process of
implementation
Strains, medium, and method details [64 , 66]
Annex 4B/1106 Microbiological examination of nonsterile products:
test for specifi ed microorganisms
In the process of
implementation
Strains, medium, method details, and result interpretation [64 , 66]
Annex 4C/1107 Microbiological examination of nonsterile products:
acceptance criteria for pharmaceutical preparations
and substances for pharmaceutical use
In the process of
implementation
Scope, Salmonella tests, and microbial acceptance criteria for
small and microdose preparations such as patches, and standards
for traditional Chinese medicines (vegetable medicines)
[64 ,66]
Annex 5/0921 Disintegration test In the process of
implementation
Apparatus, result interpretation [64e66]
Annex 6/0941 Uniformity of dosage units In the process of
implementation
Methods, result interpretation [64e66]
Annex 7/0931 Dissolution test In the process of
implementation
Methods, result interpretation [64e66]
Annex 8/1101 Sterility test In the process of
implementation
Strains, number of products to be tested, fi lter times, and quantity
of the rinsing fluid
[64 ,66]
Annex 9/0923 Tablet friability In the process of
implementation
Apparatus, notes [64e67]
Annex 10/0541 Polyacrylamide gel electrophoresis In the process of
implementation
Method details [64e66]
Annex 11/0542 Capillary electrophoresis In the process of
implementation
Method details [64e66]
Annex 12/0982 Analytical sieving In the process of
implementation
Chinese Pharmacopoeia includes the manual sieving method; the
ICH guideline includes the sonic-sifter sieving method
[64e66]
Annex 13/0993 Bulk density and tapped density of powders In the process of
implementation
No changes [64e66]
Annex 14/1145 Bacterial endotoxins test In the process of
implementation
Method description [64 , 66]
X. Xu, H. Xu, Y. Shang et al. Journal of Pharmaceutical Analysis 11 (2021) 398e404
400
quality control and the current applicability of products already on
the market in China, the newly added general technical re-
quirements are consistent with the ICH guidelines, and the revised
general technical requirements are better harmonized with the ICH
guidelines as much as possible.
Stability is one of the critical factors infl uencing competivity in
the drug market and is an important fi eld of technological inno-
vation. Guideline 9001, " Stability Testing of Drug Substances and
Preparations" , was revised according to the ICH Q1A [68 ]. The
defi nition of "signifi cant changes" in preparation quality was pro-
posed to guide manufacturers to focus on critical quality attributes.
Additionally, the requirements for the transportation of
temperature-sensitive drugs have been clarifi ed. For special prep-
arations, such as sustained- and controlled-release preparations
and inhalations, the important parameters affecting their stability
test are listed. Guideline 9101, " Validation of Analytical Methods",
was revised to be consistent with the ICH Q2 [69 ]; the contents
regarding the correction factor were deleted and the methods for
accuracy and precision were revised. The reporting, identification,
and qualifi cation thresholds for drug impurities and the decision
tree of the ICH Q3A and Q3B [70 ,71 ] were introduced in guideline
9102, " Analysis of Impurities in Drugs" . To ensure consistency with
the ICH Q3C [72 ], cumene and methyl isobutyl ketone were revised
from class 3 solvents to class 2 solvents, and triethylamine, a class 3
solvent, was added to method 0861, " Determination of Residual
Solvents" . The fl ow-through cell and reciprocating cylinder
methods were added to method 0931, " Dissolution and Drug
Release Test" . The instruments, methods, and interpretations
related to the ICH Q4B Annex 7 [73e82 ] and the research results on
specifi c preparations, such as compound ketoconazole cream and
lithium carbonate sustained-release tablets, were introduced.
Method 1101, " Sterility Tests" , was improved, based on the ICH Q4B
Annex 8, to be more instructive and practical [83 ]. The scope of
environmental monitoring, the storage and use of culture media
and strains, the culture time of the medium sensitivity test, the
number and quantity of products to be tested, and the re-
quirements of incubation and observation were revised. The bulk
density and tapped density are important functionality-related
characteristics of pharmaceutical excipients in powder form.
These densities are commonly used to calculate the Hausner ratio
and compressibility index of the powders. Referring to the ICH Q4B
Annex 13 [84e86 ], method 0993, " Bulk Density and Tapped Density
of Powders" , was added. Referring to the ICH M7 [87 ], guideline
9306, " Genotoxic Impurities Control" , was added, and the general
principles, assessment methods, and calculation methods of
acceptable intakes and limits were introduced.
However, some general testing methods in the Chinese Phar-
macopoeia 2020 edition still differ from those in the ICH Q4B
(Table 3 ). The general testing methods in the Chinese Pharmaco-
poeia were originally drafted according to the British Pharmaco-
poeia and the World Health Organization, and these general
chapter methods have a long history of use and a wide variety of
applications in China. However, the current mainstream drug
standard harmonization is based on the Pharmacopoeia Discussion
Group and the ICH. Moreover, due to limited information and the
complexity of regulatory adjustments, the information in interna-
tional standards referenced by the Chinese Pharmacopoeia is not
comprehensive, and the revisions are not timely. Despite these
challenges, harmonization with international standards is still
vigorously promoted by the Chinese Pharmacopoeia Commission
(ChPC). In October 2018, the ICH Q4 symposium was held in Beijing.
More than 20 experts from the ICH Expert Working Group and
ChPC discussed strategies for implementing ICH Q4 in China. In
2020, the ICH Q4B implementation status of the Chinese Pharma-
copoeia was added to the offi cial ICH website for the fi rst time [64].
5. Summary and prospects
The general chapters of the Chinese Pharmacopoeia 2020 edi-
tion are based on science, risk, and applicability, and refer to the
ICH guidelines. New technologies and requirements developed in
recent years were introduced to provide technical and methodo-
logical support to ensure the safety, effectiveness, and controlla-
bility of pharmaceuticals in China. The current edition will play an
active role in encouraging the application of advanced technolo-
gies, improving quality control of drugs, and strengthening the
means of drug regulation in China.
As observed from the history of other pharmacopoeias, the
development of drug standards is a process of gradual and
continuous improvement owing to the limitations of scientific
cognition. The concept of quality by design and life cycle manage-
ment will be further implemented in the general chapters of the
2025 Chinese Pharmacopoeia [88e96 ]. For example, analytical
procedure lifecycle guidelines and process analysis technologies
will be introduced, and the roles of statistical methods in data
evaluation, interpretation, and processing for the development,
validation, transfer, and verifi cation of analytical methods will be
strengthened. The system for microbiological control based on risk
assessment will also be improved.
Widely used analytical technologies, such as HPLC, GC, and
atomic spectroscopy, will be revised. Moreover, additional scienti-
fic, objective, and convenient techniques will be introduced in a
timely manner. The development of personalized microbial testing
methods for specifi c preparations and research detailing rapid
microbiological methods will be further elaborated [97e 101 ]. The
testing methods for active and toxic ingredients, exogenous pol-
lutants in crude TCMs, and the microbiological examination re-
quirements for TCM decoction pieces will continue to be improved.
In 2020, the revision of the Q4B guidelines was initiated by the
ICH. The ChPC will continue to expand its participation in the
harmonization of drug standards and actively promote harmoni-
zation/interchangeability with ICH Q4 based on validation. Other
ICH guidelines, such as the ICH Q3D, will also be harmonized.
Further, the general chapter of " Elemental Impurities Limits and
Procedures" in the Chinese Pharmacopoeia will be developed to
better assess and control elemental impurities in drugs in China.
Declaration of competing interest
The authors declare that there are no confl icts of interest.
Acknowledgments
The authors acknowledge the fi nancial support from the Chi-
nese Pharmacopoeia Commission Drug Standard Promoting Funds
and Comprehensive Reform of the Chinese Drug and Medical De-
vice Review and Approval System Funds (2015e2020).
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Endotoxin in dairy products may pose risks to consumers' health, but the effects of raw-milk total bacterial count (TBC) and thermal treatment temperature on endotoxin content of dairy products are not clear. In this study, TBC of raw milk and processed milk with different thermal treatment temperatures were detected by pilot processing experiment. We then verified it by detecting the endotoxin content of commercial dairy products. TBC was determined by conventional culture method, and the changes in alkaline phosphatase (ALP) and lactoperoxidase (LPO) activities in bovine milk before and after heat treatment were determined. The levels of fatty acids in bovine milk were detected by gas chromatography using a flame ionization detector (GC-FID). In the pilot experiment, raw milk was treated at 72, 75, 80, 85, 90, 95, 100, 105, 110, 115, and 120 ℃ for 15 s. After that, the raw milk was stored at 20 ℃ for 24 hours and 48 hours to greatly increase the TBC, and treated at 75 ℃ for 15 s and 90 ℃ for 15 s. The results show that the endotoxin content of sterilized milk increased with the increase in TBC and thermal treatment temperature. With the increase in thermal treatment temperature, the activities of ALP and LPO with endotoxin inhibition decreased. In addition, a total of 226 dairy samples were tested. The endotoxin content of 37 commercial PM milk samples were 6 EU/ml to 231 EU/ml, with 31 EU/ml to 1437 EU/ml for 40 commercial UHT milk samples, and 6 EU/ml to 9080 EU/ml for 149 reconstituted IFM samples. There was a significant difference in the endotoxin content of different dairy products (P < 0.05), and there was no significant difference between domestic and imported IFM (P > 0.05). This study found that TBC and thermal treatment temperature of raw milk could affect the endotoxin content of dairy products.
- Qing Zhu
- Ping Ye
- Fang Guo
- Zhaohua Chang
In this study, the surface of the covered stent was treated by plasma technology to introduce amino functional groups, and glutaraldehyde and heparin were successfully grafted to prepare a heparin-functionalized covered stent (HPLCS). The preparation parameters such as plasma treatment power, plasma treatment time, concentration of glutaraldehyde and heparin, and pH of heparin solution were studied in detail. The functionalized heparin covered stent can make the titer of heparin reach 1.23 ± 0.03 IU/cm ² . In animal experiments, after implantation in pigs for 6 months, the titer of heparin can still reach 0.93 ± 0.05 IU/cm ² . This work provides a good method for preparing heparin covered stent.
- Weida Chen
- Xiang Li
- Li Zeng
- Zhichuan Liu
In the current study an Allicin delivering polymeric wound dressing was formulated using thermally induced phase separation method. Allicin was loaded into Chitosan/Polyvinyl Alcohol solution at weight ratios 3 %,5 %, and 10 % and then freeze-dried. The fabricated membrane's physiochemical and biological properties were assessed using scanning electron microscopy, cell viability assay, swelling capacity, mechanical strength, surface wettability, water vapor permeability, drug release profile, in vitro degradation, hemocompatibility, porosity measurement, microbial penetration assay, Anti-oxidant assay, and protein adsorption assay. According to cell MTT assay results, Chitosan/Polyvinyl Alcohol incorporated with 5 % Allicin had the highest cell viability compared with other formulations; therefore, this formulation was chosen for treating rat model of diabetic wounds. The results showed that the rats treated with Allicin loaded dressings exhibited 93.15 ± 6.38 % of wound closure after 14 days. In addition, this group showed 47.11 ± 3.67 μm of epithelial thickness and 72.31 ± 4.28% of collagen deposition at the end of 14th post-wounding. These values were significantly greater than that of Allicin-free dressings and control group, p value < 0.05. These preliminary results suggest potential applicability of Allicin loaded Chitosan/Polyvinyl Alcohol dressing in treating diabetic wounds in clinic.
Objective: Comparison was done between high-performance liquid chromatography (HPLC) and a competitive enzyme-linked immunosorbent assay (ELISA) for detection and quantification of aflatoxin B1 (AFB1) in feed samples. The two procedures were standardized and validated before the actual experiment. Five concentrations (0, 5, 10, 20 and 30 ppb) of feed samples were used for both methods. For the HPLC technique, the samples were extracted in acetonitrile/water (90/10) solution, cleaned-up using solid phase extraction (SPE) column, and derivatized by water/trifluoroacetic acid/glacial acetic acid (35/10/5) solution before instrument analysis. The samples were extracted in 70% methanol for the ELISA technique. Results: The two tests showed very strong linearity with correlation coefficient value of > 0.99 using standard solutions. The mean recovery rate was 92.42% (with relative standard deviation (RSD) of 5.97) and 75.64% (RSD = 34.88) for HPLC and ELISA, respectively. There was no statistically significant difference in recovery rate between the two methods. There was a positive correlation (r = 0.84) between them which indicated that the two techniques can be used to detect and quantify aflatoxin B1 in feed samples. However, there were variations among replicates for the ELISA method, which shows that this method is more applicable for screening purposes.
- Nan Tie-Gui
- Hong Xiao-Xu
- X U Xin-Yi
- Yuan Yuan
The enzyme linked immunosorbent assay of aflatoxin has been adopted in Chinese Pharmacopoeia(2020 edition). Based on high-throughput screening of monoclonal antibodies technology, monoclonal antibodies that can specifically recognize the aflatoxin B_1 and the total amount of aflatoxin B_1, B_2, G_1, and G_2 in Chinese herbal medicines were prepared. By optimizing the concentration of coating antibody, enzyme-labeled antigen, and the reaction system of enzyme-linked immunosorbent assay, the enzyme linked immunosorbent assay(ELISA) were developed for detection of aflatoxins in Chinese herbal medicines, decoction pieces, and preparation of Chinese medicine. In this method, the recovery test of actual samples is 60%-120%, and the relative standard deviation is less than 15%. In addition, in view of the complicated and expensive pretreatment methods for the determination of aflatoxin in Chinese herbal medicine, we developed a highly efficient pretreatment method of liquid-liquid extraction of aflatoxin in Chinese herbal medicine without immunoaffinity column. As an effective method for the detection of aflatoxin, the ELISA can effectively reduce the aflatoxins testing cost of traditional Chinese medicine, and promote the detection ability at earlier stages of production, and strengthen the quality supervision of traditional Chinese medicine.
Animal feed is a potential route for contaminants like mycotoxins to enter into the human food chain. Hence, close monitoring is fundamental and should be performed with adequate analytical methods. In this study, a commercial ELISA kit for aflatoxin B1 detection in feed corn samples, RIDASCREEN® Aflatoxin B1 30/15, was assessed by the evaluation of some performance parameters. Basically, the following results were achieved: limit of detection of 1.1 μg/kg and limit of quantification of 2.5 μg/kg; repeatability of 9.3%; 18.0% of intermediate precision; trueness of 101.8%; relative expanded uncertainty of ± 0.46 μg/kg; cut-off value of 14 μg/kg and a very low rate of false suspect results, for screening. In addition, specific requirements for the appliance of this kit as a screening method at the concentration of 20 μg/kg were also determined. Lastly, corn samples were analysed with the kit and with an LC-MS/MS method, and the results were compared. Among the 40 corn samples analysed, only one sample was contaminated with a quantifiable aflatoxin B1 content. The results obtained with the ELISA kit were confirmed by LC-MS/MS. Graphical abstract
- Hairong He
- Fei Gao
- Yonghong Zhang
- Xiaoke Zheng
The behaviour of residues of tebuconazole, prochloraz, and abamectin in rehmannia during rehmannia decoction processing was systemically assessed. The pesticides were determined by ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) after each processing step including washing, steaming and drying, carbonising, and boiling. Results showed that the pesticide residues significantly decreased after the steps of washing, carbonising, and boiling. Washing reduced pesticide residues by 41.2%–60.0%; carbonising reduced pesticides by 27.1%–71.1% in both prepared rehmannia and unprepared rehmannia. After boiling, the concentrations of tebuconazole and prochloraz were 0.0002–0.0022 mg kg⁻¹ in decoctions. Abamectin was not detected in rehmannia after carbonising, and it was not detected in decoctions either. The processing factors (PFs) were less than 1 during food processing, indicating that the full set of processing can reduce the residues of tebuconazole, prochloraz, and abamectin in rehmannia decoction.
- Nathan D. Rudd
- Mikhail Reibarkh
- Rui Fang
- William P. Forrest
Injectable sustained release dosage forms have emerged as desirable therapeutic routes for patients that require life-long treatments. The prevalence of drug molecules with low aqueous solubility and bioavailability has added momentum towards the development of suspension based long-acting parenteral (LAP) formulations; the previously undesirable physicochemical properties of BCS Class II/IV compounds are best suited for extended release applications. Effective in vitro release (IVR) testing of crystalline suspensions affirms product quality during early-stage development and provides connections with in vivo performance. However, before in vitro-in vivo correlations (IVIVC) can be established, it is necessary to evaluate formulation attributes that directly affect IVR properties. In this work, a series of crystalline LAP nanosuspensions were formulated with different stabilizing polymers and applied to a continuous flow-through (USP-4) dissolution method. This technique confirmed the role of salt effects on the stability of polymer-coated nanoparticles through the detection of disparate API release profiles. The polymer stabilizers with extended hydrophilic chains exhibited elevated intra-polymer activity from the loss of hydrogen bond cushioning in dissolution media with heightened ionic strength, confirmed through 1D 1H NMR and 2D NOESY experiments. Thus, steric repulsion within the affected nanosuspensions was limited and release rates decreased. Additionally, the strength of interaction between hydrophobic polymer components and the API crystalline surface contributed to suspension dissolution properties, confirmed through solution- and solid-state spectroscopic analyses. This study provides a perspective on the dynamic interface between the crystalline drug and aqueous micro-environment during dissolution.
The aim of this study was to investigate the impact of mycotoxin binders on the determination of aflatoxins in maize and maize gluten using various analytical methods, including ELISA, HPLC and LC-MS/MS. Three types of commercially available mycotoxin binders, yeast cell wall, mineral, and a mixture of mineral and bacterium, were investigated at inclusion levels of 0.1%, 0.2% and 0.4%. The binders were added to maize and maize gluten contaminated with aflatoxins at concentrations between 6.9 and 26.7 μg kg⁻¹. The samples were analysed and the values were compared with corresponding controls (samples without binders) using ANOVA. The yeast cell wall binder had no significant effect (p=0.05) on the concentration of aflatoxins measured in either maize or maize gluten at any of the three inclusion levels, regardless of which analytical method was used. The mineral binder and the mixed mineral and bacterium binder had no significant effect (p=0.05) on the measured aflatoxin concentrations in either maize or maize gluten at any of the three inclusion levels when analysis was conducted using LC-MS/MS. Inclusion of these binders resulted in significant lower (p<0.01) detection of aflatoxins in both maize and maize gluten when analysis was conducted using ELISA; the effect was dose-dependent. They also resulted in significant lower detection of aflatoxins in maize extracted by methanol/water (70/30 v/v) (p<0.0001) and in maize gluten extracted by acetonitrile/water (80/20 v/v) (p<0.05) when analysis was conducted using HPLC. However, neither the mineral binder nor the mixed mineral and bacterium binder had significant effects (p=0.05) on aflatoxin concentrations measured in maize using HPLC, when extracted by acetonitrile/water (80/20 v/v). The study demonstrated that mycotoxin binders could result in underestimation of the levels of aflatoxin contamination, depending on the nature of the binder, the extraction solvent used in the analytical method, and the composition of tested sample.
- Jinjing Xiao
- Xin Xu
- Fan Wang
- Haiqun Cao
The safety of Traditional Chinese Medicine (TCM) is of concern worldwide. Herein, Paeoniae Radix Alba, Chaenomelis Fructus and Moutan Cortex, representing three medicinal components, were subjected to toxicological analysis to investigate possible pesticide contamination. Exposure using a point estimate model identified 47 residues that were simultaneously validated by the QuEChERS-UPLC-MS/MS method, which is sufficiently reliable for measuring residue concentrations. Of the 313 samples tested, 94.57% contained pesticide residues, with concentrations ranging from 0.10 to 1199.84 μg kg⁻¹, of which >83.17% contained 4–15 different residues. Carbendazim was the most frequently detected pesticide (>85%), and procymidone, pendimethalin and phoxim were also abundant (median concentration = 15.33–623.12 μg kg⁻¹). Risk assessment based on the hazard quotient/hazard index (HQ/HI) approach revealed that exposure to pesticide residues in all three TCMs (95th percentile) were far below levels that might pose a health risk. However, insecticides contributed to cumulative exposure, especially phoxim, and worryingly, several banned pesticides were detected. The results are of theoretical and practical value for evaluating the safety TCMs, and could improve their quality and safety.
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![Sara Valentini]()
- Giovanna Santoro
- Federica Baffetta
- Barbara Capecchi
Pyrogen content is one of the critical quality attributes impacting the safety of a product, and there is an increasing need for assays that can reliably measure this attribute in vaccines. The Limulus amebocyte lysate (LAL) assay and the rabbit pyrogen test (RPT) are the canonical animal-based pyrogen tests currently used to release vaccines; however, there are several drawbacks associated with these tests when applied to Bexsero, intrinsically pyrogenic product, containing a meningococcal Outer Membrane Vesicle component. While the RPT, as applied to Bexsero at its given dilution, ensures safe vaccine, it is highly variable and prone to false positive results. On the other hand, the LAL assay although quantitative, can detect only endotoxin pyrogens and is not sufficient for monitoring the safety of Bexsero, which contains both LPS and non-endotoxin pyrogens. Being aware of these limitations of the RPT and LAL when applied to Bexsero, the Monocyte Activation Test (MAT) which is sensitive to both endotoxin and non-endotoxin based pyrogens has been developed as an alternative pyrogen test. Here, the development and the validation of a MAT assay adapted from the European pharmacopoeia for Bexsero, is described. The MAT assay is then used for monitoring the safety and consistency of Bexsero vaccines at release, providing great advantages in terms of reduced variability with respect to RPT, reduction of animal use, in line with the 3Rs principle concerning the protection of animals and faster time to market. In addition the correlation of the MAT to the RPT has been demonstrated supporting the replacement of the in vivo method and the potential application of the assay to other intrinsically pyrogenic vaccines.
- Tomomi Yamasaki
- Shiro Miyake
- Natsuki Sato
- Takaho Watanabe
A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) was developed for the determination of total amount of aflatoxin B1, B2, G1 and G2 (AFB1, AFB2, AFG1 and AFG2), using a mouse monoclonal antibody that shows similar reactivity to each of these AFs. The working range of the developed dc-ELISA was 50–230 pg/mL for AFB1, 50–270 pg/mL for AFB2, 60–390 pg/mL for AFG1 and 65–700 pg/mL for AFG2. The recovery of AFs from spiked roasted peanuts was 98％. Further, when 4 samples actually contaminated with AFB1, AFB2, AFG1 and AFG2 were examined, the results of dc-ELISA were highly correlated with the values assigned by the Food Analysis Performance Assessment Scheme. The developed dc-ELISA appears to be suitable for the determination of total AFs at concentrations around the maximum permitted level (10 μg/kg for all foods) in Japan.
- Ilgaz Akseli
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![Jon Hilden]()
- Jeffrey M. Katz
- John C. Strong
The bulk properties of a powder are dependent on the preparation, treatment, and storage of the sample, that is, how it was handled. The particles can be packed to have a range of bulk densities and, moreover, the slightest disturbance of the powder bed may result in a changed bulk density. Thus, the bulk density of a powder is often difficult to measure with good reproducibility and, in reporting the results, it is essential to specify how the determination was made. In this article, we measured the bulk density, tapped density, and calculated the Hausner ratio of commonly used excipients with similar tapped density testers and followed the United States Pharmacopeia 30-National Formulary 25-S1 testing procedure. Based on the analysis, within lot and lot-to-lot variability and the relative errors for bulk density, tapped density, and Hausner ratio were found to be acceptable. Lot-to-lot differences were generally not measurable using this test as they were found to be within the variability of the test. The results also indicated that there was no statistically significant bias between sites for tapped density and Hausner ratio, but there was a marginally significant bias in the bulk density data set.
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Source: https://www.researchgate.net/publication/351752805_Development_of_the_Chinese_Pharmacopoeia_2020_Edition_General_Chapters_A_Review