Ongoing procedure performance verification (OPPV) of an analytical procedure is the process of ensuring it continues to meet its intended use after completion of validation. On the basis of previous studies, this paper further discussed the indicators of ongoing procedure performance verification (including system suitability indicators and reported values, etc.) and the implementation of analysis tools (control charts), and demonstrated the specific operation steps of ongoing procedure performance verification with examples. It is hoped that this paper will provide new ideas for the accurate and standardized procedure verification in the pharmaceutical field, especially in the enterprises and regulatory departments.

Objective: To establish an HPLC method of the content and related substances of compound amino acid injection(3AA). Methods: RP-HPLC was adopted to determine compound amino acid injection(3AA), combining with the use of two-dimensional column switching-LC/MSⁿ method was applied to separate and identify the impurities. The determination was performed on Capcell PAK AQ C₁₈(250 mm×4.6 mm, 3 μm) column with 0.2 mol·L^-1 sodium dihydrogen phosphate solution (adjusted pH to 2.8 with phosphoric acid) -acetonitrile (98∶2) as mobile phase at the flow rate of 1.0 mL·min^-1. The column temperature was 40 ℃, and the detection wavelength was 210 nm. And the injection volume was 20 μL. The LC/MSⁿ method was performed on a Thermo Accucore AQ C₁₈ (100 mm×4.6 mm, 2.6 μm) column with 0.1% formic acid solution as mobile phase A, 0.1% formic acid solution-acetonitrile as mobile phase B, at a flow rate of 0.4 mL·min^-1, and at a column temperature of 40 ℃. The mass spectrometry conditions were performed using an ESI ionisation source in the positive-ion scanning mode with a scan range of m/z 100-1 000, and the secondary mass spectrum was carried out by data-dependent scanning. Results: The related substances were completely separated from the main constituents in RP-HPLC. The standard curve of valine was linear over the range of 1.263-5.050 mg·mL^-1, with the average recovery of 99.0% (n=9). The standard curve of isoleucine was linear over the range of 1.350-5.402 mg·mL^-1, with the average recovery of 99.4%(n=9). The standard curve of leucine was linear over the range of 1.647-6.588 mg·mL^-1, with the average recovery of 99.5%(n=9). The main impurities in the three batches of samples were all process impurities introduced from the raw materials, with methionine content of 4.344 μg·mL^-1, 3.751 μg·mL^-1, 4.503 μg·mL^-1, respectively, phenylalanine content of 4.636 μg·mL^-1, 4.889 μg·mL^-1, 4.753 μg·mL^-1, respectively. The maximum single impurity contents were 0.01%, 0.02% and 0.01%, respectively. Conclusion: The method is proved by the methodology validation that it can be used for the quality control of compound amino acid injection(3AA).

Althought mRNA vaccines have been developed for nearly 30 years, but due to certain technical bottlenecks in production, stability, and reactivity, the development of mRNA vaccines has been relatively slow. During the epidemic of COVID-19, mRNA vaccine had been fully verified its effectiveness and safety for preventing infectious diseases. Here, this article summarized the key quality control attributes and related requirements of mRNA vaccine products for preventing infectious diseases by reviewing the quality control guidance documents of WHO and drug regulatory and standardization agencies, aiming to provide reference for the quality control of mRNA vaccine products for preventing infectious diseases in China.

Objective: To establish an ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) for the simultaneous determination of daidzein, formononetin, genkwanin, quercetin, rutin, apigenin, kaempferol, luteolin and puerarin in Tongfu Jingyaotong tincture. Methods: The Waters ACQUITY UPLC HSS T3 Column (100 mm×2.1 mm,1.8 μm) was used with methanol and 0.1% formic acid (gradient elution) as the mobile phase at a flow rate of 0.3 mL·min^-1. Column temperature was 45 ℃, and injection volume was 2.0 μL. Electrospray ion source was adopted with positive and negative ion modes and multi-reaction monitoring and acquisition. Results: Apigenin, genkwanin and daidzein had good linearity in the range of 0.2-1 000 ng·mL^-1, puerarin, rutin, luteolin, kaempferol and formononetin in the range of 0.2-2 000 ng·mL^-1, and quercetin in the range of 1.0-5 000 ng·mL^-1. The resolution was good, and the RSDs of precision, repeatability, reproducibility and stability test were all below 5% (n=6). The average recoveries ranged from 82.7% to 111.8% with RSDs (n=6) of 2.0%-5.4%. The contents of daidzein, formononetin, genkwanin, quercetin, rutin, apigenin, kaempferol, luteolin and puerarin in 6 batches of Tongfu Jingyaotong tincture were 8.46-17.97 μg·mL^-1, 0.62-1.67 μg·mL^-1, 0.03-0.08 μg·mL^-1, 1.34-2.22 μg·mL^-1, 0.70-1.14 μg·mL^-1, 0.48-0.99 μg·mL^-1, 0.20-0.90 μg·mL^-1, 0.10-0.16 μg·mL^-1 and 167.95-227.75 μg·mL^-1, respectively. Conclusion: This method has strong specificity, wide linear range, accuracy and efficiency, and can detect the contents of 9 active ingredients in Tongfu Jingyaotong tincture at the same time, which offers an effective quality control method for Tongfu Jingyaotong tincture.

Objective: To compare the quality of wild and cultivated Arnebiae Radix,using macroscopic investigation and chemometric analysis of the different components in wild and cultivated Arnebiae Radix from three different habitats. Methods: Wild and cultivated Arnebiae Radix were collected and their macroscopic features were compared. Using the ACQUITY UPLC BEH C₁₈ (2.1 mm×100 mm, 1.7 μm) column, with acetonitrile-0.05% formic acid water as the mobile phase, the contents of D-shikonin, acetylshikonin, β-acetoxyisovalerylshikonin, isobutyrylshikonin, β,β’-dimethylacrylalkannin and isovalerylshikonin in 48 batches of wild and cultivated Arnebiae Radix were determined. The detection wavelength was 275 nm and the flow rate was 0.2 mL·min^-1. PCA and OPLS-DA were performed to reveal the differential components of wild and cultivated Arnebiae Radix. Results: There were great differences in macroscopic features of wild and cultivated Arnebiae Radix, and the linear relationship between the contents of six naphthoquinone components was good. The correlation coefficients were above 0.999, the average recovery rates were 93.4%-102.9%, and the RSDs were less than 3.0%. The contents of six components in different batches of wild and cultivated Arnebiae Radix were quite different, and the contents of D-shikonin and acetylshikonin in wild products were significantly higher than those in cultivated products, indicating that there were still certain differences between wild products and cultivated products. The PCA model established could distinguish wild products and cultivars, and two differentiating components in wild products and cultivars were revealed by OPLS-DA, namely isobutyryl shikonin, β,β’-dimethylacrylalkannin. Conclusion: By comparing the core size, cork curl degree and specific odor of wild and cultivated products, the two can be identified. The established content determination method is repeatable, specific, stable and feasible. The differential components in wild and cultivated Arnebiae Radix in three different regions are identified, which provides a basis for the quality control of Arnebiae Radix and provides ideas for expanding the source of Arnebiae Radix.

The change of analytical procedure, a part of the entire lifecycle of a method, is increasingly valued with the emergence of new technologies, the increasing demand for product quality, 3R, environmental protection, and cost reduction requirements. This paper refers to the latest progress in the study of counterpart law at home and abroad, systematically classifies changes of analytical procedures from the perspective of methodology, and discusses the differences between different types of changes and the evaluation criteria and implementation approaches of analytical procedures after procedure changes.

Objective: To establish a stable and reliable gas chromatography-tandem mass spectrometry method for the determination of N-nitrosodimethylamine (NDMA) in metformin hydrochloride sustained-release tablets, and to optimize extraction conditions by investigating the effects of isopropyl alcohol and thioglycerin on the stability of the tested solution. Methods: Separation was achieved on AB-InoWax capillary column (30 m×0.25 mm×0.25 μm) with polyethylene glycol as the stationary phase. Electron ion(EI) source and multiple reaction monitoring (MRM) mode were used. Quantitative determination was performed by both external standard and internal standard. Results: The addition of thioglycerin could significantly improve the stability of the test solution. NDMA showed good linearity within the concentration range of 0.25-50 ng·mL^-1(r>0.999). The detection limit of the method was 0.1 ng·g^-1 and the quantification limit of the method was 0.2 ng·g^-1. The average recoveries (n=9) were 97.3% and 94.9% while using external standard and internal standard, respectively. Precision, repeatability and stability were good with RSD less than 8%. Ninety batches of metformin hydrochloride sustained release tablets were tested. NDMA content in all detected samples were all less than 30% of the acceptable limit set by the National Medical Products Administration and FDA. Conclusions: This method shows satisfactory sensibility, specificity, accuracy, stability and durability, which is suitable for quantitative analysis of NDMA in metformin hydrochloride sustained-release tablets, providing technical support for the quality and safety of related products.

This article focuses on the key experimental operations in the study about the applicability of microbial limit testing methods for drugs, summarizing literature and experience from the preparation of test solution, design of methods for microbial counting and control bacteria inspection, operation of microbial recovery test, bacterial liquid concentration counting, and operational methods for controlling bacterial & fungal purity. Emphasis was placed on the preparation methods of water-soluble test samples with strong antibacterial effects, non-oil and fat test samples that were not easily soluble and dispersed, and the test solution for oil and fat test samples. Detailed introduction of the sequential experimental plan for methods of aerobic bacterial count, mold & yeast count, and control bacterial inspection were introduced. Four operational methods for adding bacteria about membrane filtration were explained, and the impact of different ways of adding bacteria on the results of the method was analyzed. Summarized three counting methods for bacterial & fungal concentrations. Detailed sharing of operational experience on controlling the purity of bacterial solution. Summarized the current status about research on the applicability of microbial limit testing methods of drugs. Four suggestions for the future development of research on the applicability of microbial limit testing methods for drugs have been proposed:(1) Unify the operation about bacteria adding method in membrane filtration,scientific and reasonable;(2)Strengthen supervision of pharmaceutical production enterprises and review of microbial limit testing methods for drugs;(3)Unify the microbial limit testing methods for the drugs of the national sampling plan,and gradually collect and bind into a book;(4)Strengthen the research on the applicability of microbial limit testing methods for excipients.

Objective: To study and establish a method based on gas chromatography and chemometrics techniques for distinguishing Artemisiae Argyi Folium and its adulterants Artemisiae Mongolica Folium. Methods: Gas chromatography method was established with Agilent HP-5 19091J (30 m×0.32 mm, 0.25 μm) as chromatographic column, and hydrogen flame ion detector (FID) as detector. After the chemical composition of 21 chromatographic peaks in the chromatogram were identified, and the peak area data of the 21 chromatographic peaks in 29 batches of samples were determined. Similarity analysis, correlation analysis, cluster analysis, principal component analysis and orthogonal partial least squares-discriminant analysis were applied to analyze the chromatographic data. Results: The results of chemometric analysis indicated that tpeak 20 (chamazulene), peak 3 (1,8-cineole) and peak 19((1S,8aα)-decahydro-1,4aβ-dimethyl-7β-isopropenyl-1-naphthol) were the differential characteristic chromatographic peaks between Artemisiae Argyi Folium and its adulterants Artemisiae Mongolica Folium. The ratios of the peak areas of peak 3 to peak 20 were in the ranges of 54.50-348.39 and 0.16-0.87 respectively, and the ratios of the peak areas of peak 19 to peak 20 were in the ranges of 18.55-128.46 and 0.01-0.14 respectively. These significant differences could be used for the identification of Artemisiae Argyi Folium and its adulterant Artemisiae Mongolica Folium. Conclusion: The research findings can be used for the identification of Artemisiae Argyi Folium and its adulterant Artemisiae Mongolica Folium, and these have certain reference significance for the research and analysis of Artemisiae Argyi Folium and related drugs.

Objective: To establish a fast, accurate and sensitive method by UPLC-DAD-Q-TOF-MS/MS for the detection of 20 kinds of illegally added chemicals such as azilsartan,nifedipin, lacidipine、nebivolo in anti-hypertensive health foods. Methods: Waters HSS T3 C₁₈(100 mm×2.1 mm,1.8 μm ) column was used with mobile phase of 0.01 mol·L^-1 ammonium acetate aqueous solution-methanol and gradient elution(0 min, 5%A;0-8 min, 5%A→95%A;8-11 min,95%A;11-11.2 min,95%A→5%A;11.2-12 min,5%). The flow rate was 0.3 mL·min^-1. The injection volume was 1 μL. ESI⁺,ESI^- scan and MS/MS mode were applied. Qualitative and quantitative analyses were carried out by UPLC-DAD-Q-TOF-MS/MS. Results: The method was validated. A good linear relationship was showed in the concentration of 1-50 μg·mL^-1,(RSD<3%)and the recovery was obtained in range of 80.5%-98.7%. Illegally added chemical drugs were found in 15 batches of samples tested by this method. Two or more types of drugs were detected in 14 batches of samples. Torsemide, candesartan cilexetil, lacidipine were detected in 9 samples. Lacidipine and nebivolol were found in one of the sample. Conclusion: This method is fast, accurate and sensitive with a high degree of separation. It can be used as a powerful tool for the fast detection of illegally added chemicals in anti-hypertensive health foods.

Objective: To establish an innovative analytical method based on high resolution sampling two-dimensional chromatography (HiRes 2D-LC) for determination of the content of vitamin D₃ in vitamin D drops. Methods: Two-dimensional liquid chromatography was used. Thermo HYPERSIL Gold Silica (100 mm×2.1 mm, 1.9 μm) column was used in the first dimension with n-hexane-n-amyl alcohol (996∶4) as mobile phase. The flow rate was 0.2 mL·min^-1. The samples were injected and tested at the wavelength of 265 nm. The column temperature was 40 ℃. In the second dimension liquid chromatography, ShimPack Velox Hilic (50 mm×2.1 mm, 2.7 μm) was used as the column with n-hexane-n-pentanol-isopropanol (98∶1∶1) as the mobile phase. The flow rate was 0.5 mL·min^-1. The samples were injected and tested at the wavelength of 265 nm. The column temperature was 40 ℃. A six-position 14-way valve and was equipped with 2 multi-center cutting valves was equipped to make multiple consecutive cuts of the pre-vitamin D₃ peak and vitamin D₃ peak. Results: The calibration curves showed a good linearity at the range of 1.018 4-5.092 mg·mL^-1(r≥0.999 8). The precision test showed the RSDs of the peak area of pre-vitamin D₃ and vitamin D₃ were 0.95% and 0.40%, respectively. The repeatability test showed the RSD of vitamin D₃ content was 0.41%. The average recovery rate (n=9) was 101.4%. The test solution was stable at 4 ℃ and 10 ℃ for 12 h, and the RSDs were 0.58% and 0.66%, respectively. The contents of vitamin D₃ in the samples of vitamin D drops measured by this method were 100.4%, 101.6%, 100.9%, 101.6%, 102.7% and 101.6%, which was basically consistent with the results measured by the fourth method in General Chapter 0722 of ChP 2020 Vol Ⅳ. Conclusion: This method has a good specificity and high sensitivity to accurately determine the content of vitamin D₃ in vitamin D drops.

Objective: To establish the RP-UPLC-PDA method for simultaneous determination of triptolide, triptonide, triptophenolide, wilforine, wilforlide A and celastrol in Tripterygii Radix. Methods: Tripterygii Radix were extracted with ethyl acetate and the extracts were dissolved and separated by methanol. The six components were determined by RP-UPLC-PDA method. The chromatographic column was Acquity^TM UPLC BEH C₁₈ column (100 mm×2.1 mm, 1.7 μm), the column temperature was 30 ℃, the injection volumn was 2 μL, the flow rate was 0.4 mL·min^-1 and the mobile phase was acetonitrile (A)-0.1% formic acid (B). Results: The linear relationship of six components was good (0.999 2≤r≤0.999 7) in the concentration ranges. The average recoveries were 99.2%-103.1% and RSDs were 1.2%-2.9%. The contents in 10 batches of Tripterygii Radix from different habitat were determined. The results showed that the contents of Tripterygii Radix in prepared pieces from different producing areas were different. The highest content of triptolide was 140.2 μg·g^-1, and the lowest content was 103.2 μg·g^-1. The highest content of triptonide was 224.7 μg·g^-1 and the lowest content was 112.2 μg·g^-1. The highest content of triptophenolide was 306.7 μg·g^-1 and the lowest content was 189.6 μg·g^-1. The highest and lowest contents of wilforine were 283.2 μg·g^-1 and 211.2 μg·g^-1. The highest content of wilfhabitat was 31.2 μg·g^-1 and the lowest content was 16.8 μg·g^-1. The highest content of celastrol was 87.6 μg·g^-1, and the lowest content was 52.1 μg·g^-1. Conclusion: The RP-UPLC-PDA method can simultaneously determine six components in Tripterygii Radix. The method is reliable and stable, which is suitable for quantitative analysis and determination of Tripterygii Radix.

Objective: To establish a quantitative analysis of multi-components by single marker (QAMS) for simultaneous determination of 7 flavanoids (5-hydroxy-6, 7, 3’, 4’-tetramethoxyflavone, apigenin, hispidulin, kaempferol, jaceosidin, eupatilin and casticin) in Artemisiae Argyi Folium. Methods: The HPLC system consisted of the Agilent ZORBAX SB-C₁₈ column (150 mm×4.6 mm, 5 μm) column with gradient elution of acetonitrile and 0.2% phosphoric acid as the mobile phase at a flow rate of 1.0 mL·min^-1, a detection wavelength of 350 nm, and a column temperature of 30 ℃. Eupatilin was selected as the internal reference substance, the relative correction factors between eupatilin and the other 6 flavanoids were established, and the contents of these 7 constituents in samples were calculated to realize QAMS. At the same time, compared with the external standard method to verify the accuracy and feasibility of the QAMS method. Results: Within a certain linear range, the relative correction factors between eupatilin and 5-hydroxy-6, 7, 3’, 4’-tetramethoxyflavone, apigenin, hispidulin, kaempferol, jaceosidin as well as casticin were 0.958, 1.387, 1.000, 0.950, 0.957 and 1.297, respectively (RSDs of RCFs were less than 2.0%). The contents of 5-hydroxy-6, 7, 3’, 4’-tetramethoxyflavone, apigenin, hispidulin, kaempferol, jaceosidin, eupatilin, casticin in 20 batches of Artemisiae Argyi Folium were 0.031 4-0.623 5 mg·g^-1, 0.000 9-0.092 6 mg·g^-1, 0.020 6-0.170 7 mg·g^-1, 0.011 0-0.184 7 mg·g^-1, 0.011 7-0.864 0 mg·g^-1, 0.253 2-2.555 0 mg·g^-1 and 0.015 6-0.250 7 mg·g^-1, respectively. Conclusion: Using eupatilin as the internal reference, QAMS method for 7 flavanoids is established. The method is accurate and reliable, and can be used for quality control and quantitative analysis of Artemisiae Argyi Folium.

Objective: To establish an HPLC characteristic chromatogram of Xifeng Huoluo capsules, and to determine the contents of gastrodin, amygdalin, strychnine, hydroxysafflower yellow A, brucine, paeoniflorin, ferulic acid, salvianolic acid B, cryptotanshinone, tanshinone Ⅰ, and tanshinone ⅡA. Methods: Extraction of 75% methanol was analyzed by a Venusil MP C₁₈ chromatographic column (250 mm×4.6 mm, 5 μm) using 0.1% phosphoric acid (A) -acetonitrile (B) as the mobile phase with gradient elution at a flow rate of 1.0 mL·min^-1. The column temperature was 30 ℃. The detection wavelengths were variable. The characteristic chromatograms of 10 batches of Xifeng Huoluo capsules were evaluated by similarity evaluation, and the eleven identified index components were quantitatively determined. Results: There were thirty-one common peaks in the characteristic chromatograms of eleven batches of samples and the similarities were all above 0.99. The common peaks could ascribed to twelve medicinal materials, and eleven of them were identified. Eleven components showed good linearity within their respective ranges (r≥0.999 4), and the average recoveries were 99.3%-103.0% with RSDs of 1.5%-2.4%. Conclusion: The established method has high sensitivity and strong specificity. The HPLC characteristic chromatogram combined with multi-component quantitative determination can fully reflect its inherent quality and can be used for the quality control of Xifeng Huoluo Capsules.

Objective: To determine the color of Arnebiae Radix, and the contents of six main purple pigment components (acetylshikonin, β-acetoxyisovalerylalkannin, deoxyshikonin, isobutylshikonin, β, β’-dimethylacrylalkannin and isovalerylshikonin) in Arnebiae Radix, and to study on the correlation between the color of Arnebiae Radix and the contents of six main purple pigment components. Methods: The L, a, and b values of the sample powder were determined using a spectrophotometer to characterize the color of Arnebiae Radix. The International Commission on Illumination (CIE) had developed a Lab color model, which was a digital description of human vision. A higher L value indicated greater brightness, a higher a value indicated redness and a lower a value indicates greenness, and a higher b value indicated yellowing and a lower b value indicates blueness. The contents of purple pigment components were determined using high performance liquid chromatography (HPLC), and the correlation between L, a, and b values and the contents of six main purple pigments was calculated using SPSS software. Results: The contents of acetylshikonin in 135 batches of samples ranged from 0.01% to 3.39%. The contents of β-acetoxyisovalerylalkannin ranged from 0.00% to 1.95%. The contents of deoxyshikonin ranged from 0.00% to 0.23%. The contents of isobutylshikonin ranged from 0.01% to 1.13%. And the contents of isovalerylshikonin ranged from 0.02% to 2.88%. The contents of β, β’-dimethylacrylalkannin ranged from 0.01% to 2.17%. There was a significant negative correlation between the contents of acetylshikonin, β-acetoxyisovalerylalkannin and isobutylshikonin and the L (black_white) chromaticity value of Arnebiae Radix, with a Spearman correlation coefficients between -0.138 and -0.222. The chromaticity value of a (red_green) was related to the five components other than acetylshikonin, which were β-acetoxyisovalerylalkannin, deoxyshikonin, isobutylshikonin, β, β’-dimethylacrylalkannin, and isovalerylshikonin, with a spearman correlation coefficients between 0.176 and 0.355; b (blue_yellow) chromaticity value was related to the five components other than β-acetoxyisovalerylalkannin, which were acetylshikonin, deoxyshikonin, and isobutylshikonin, β, β’-dimethylacrylalkannin. β, β’-dimethylacrylalkannin was positively correlated with a coefficient of 0.290, and negatively correlated with the other four components with a coefficients between -0.325 and -0.633. Conclusion: It is recommended that the assay limits of Arnebiae Radix be revised to β, β’-dimethylacrylalkannin not less than 0.30% and isovalerylshikonin not less than 0.29%.

Objective: To provide reference for quality control and authenticity identification of Arnebiae Radix medicinal materials and decoction pieces in the market. By studied on the ITS2 sequences’ characters of imported Arnebiae Radix, based on DNA barcoding and PCR-RFLP technologies. Methods: The ITS2 region was selected as the DNA barcode sequence for comparison and identification of imported Arnebiae Radix and reference medicinal materials. The ITS2 sequences of imported Arnebiae Radix from different sources with reference medicinal materials were compared based on DNA barcoding and PCR-RFLP technologies. Results: After the restriction endonucliase AluI enzyme digestion, the agarose-gel electrophoresis results of 39 imported Arnebiae Radix samples showed that, only DH3 had bands at around 500 bp, and none bands between 100 bp and 300 bp. And the results of other imported Arnebiae Radix samples had two or three obvious bands between 100 bp and 300 bp. The ITS2 sequences of imported Arnebiae Radix samples were compared with the reference medicinal materials, among which DH3 had the largest differences of 15 bases compared to the reference medicinal materials, the ITS2 sequence of F2 was same to the reference medicinal materials, and other imported Arnebiae Radix samples had 1-9 bases difference compared to the reference medicinal materials. The clustering results showed that the imported Arnebiae Radix sample DH3 was clearly distinguished from other imported Arnebiae Radix samples and reference medicinal materials which was in a single branch. There were 14 samples, which were clustered together with the reference medicinal materials in one branch with support rate ≥50%. Conclusion: The ITS2 region is selected to compare the similarities and differences of ITS2 sequences between imported Arnebiae Radix samples and reference medicinal materials based on DNA barcode and PCR-RFLP technologies, which provids a reference for effective identification of Arnebiae Radix medicinal materials and decoction pieces, and a strong guarantee for market supervision of Arnebiae Radix medicinal materials.

Objective: To establish a rapid and accurate ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the detection of dextromethorphan and one metabolite dextrorphan in hair. Methods: The hair sample containing dextromethorphan and dextrorphan were extracted with methanol containing internal standard proadifen hydrochloride(SKF_525A). The extract was filtered with 0.22 μm organic filter membrane and detected by UPLC-MS/MS. All components were separated by an ACQUITY UPLC HSS T3 column (100 mm×2.1 mm, 1.8 μm), using a gradient elution procedure consisting of 0.2% formic acid (10 mmol·L^-1 ammonium formate) and acetonitrile, at a flow rate of 0.3 mL·min^-1, and the column temperature was room temperature. Positive electrospray ionization was performed using multiple reaction monitoring mode (MRM). Results: The linear relationships of dextromethorphan and dextrorphan were good in the range of 1-100 ng·mL^-1. The linear equations were Y=1.349 49X-0.020 80 (r=0.998 8) and Y=0.775 10X-0.013 87 (r=0.999 1), respectively. The detection limit and quantitation limit were 0.010 ng·mL^-1 and 0.025 ng·mL^-1 respectively. Their recoveries ranged from 97.0%-104.8%. The intra-day RSD and inter-day RSD were 1.5%-3.9% and 2.1%-5.5%, respectively. The method was applied to cases, and the results showed that dextromethorphan and dextrorphan were detected in the hair of 6 abusers.Conclusion: This method is simple and sensitive enough to be applied to detect dextromethorphan and one metabolite dextrorphan in hair.

Objective: To optimize the content determination method of morphine sulfate suppositories, which had high toxicity of extraction solvent and poor specificity and generality of chromatographic conditions. Methods: The optimization of the preparation method for the test solution was to prepare the test solution by ice bath freezing and n-heptane extraction, and to compare the content determination results of the test solution prepared by the United States Pharmacopeia method (extracted with trichloromethane). The optimization of chromatographic conditions for content determination was based on the comprehensive comparison of United States Pharmacopeia, British Pharmacopeia and YBH specification. Symmetry C₁₈ column (150 mm×4.6 mm, 5 μm) was finally used. The mobile phase was 0.01 mol·L^-1 potassium dihydrogen phosphate solution containing 0.202% sodium heptane sulfonate (containing 0.1% triethylamine, adjusted to pH 2.5 with phosphoric acid) -methanol (70∶30). The flow rate was 1.0 mL·min^-1. The column temperature was 30 ℃. The detection wavelength was 284 nm. The injection volume was 20 μL. Results: The content measured by ice bath freezing method was relatively low, while the content measured by n-heptane extraction and trichloromethane extraction was basically consistent, indicating that n-heptane could be used as extraction solvent instead of halogenated alkane. The linear range of morphine sulfate was 2.12-105.94 μg·mL^-1(r=0.999 9), and the LOQ was 10.60 ng. The RSD values of system precision, repeatability, intermediate precision and stability were all less than 2%. The average recoveries were 98.6%-99.6% and RSD was 0.21%-0.66%. After forced degradation, the main peak of the tested solution could be separated from the degraded impurities, with good specificity. By changing the flow rate, column temperature, pH value, mobile phase ratio and column brand, the main peak could be well separated from each impurity peak, with good robustness. Conclusions: This study optimized the determination method for morphine sulfate suppositories. The preparation of the test solution used n-heptane instead of the highly toxic haloalkane, and meanwhile improved the chromatographic conditions, enhanced the environmental protection, specificity and applicability of the method. It can be used for the content determination of morphine sulfate suppositories.

Objective: To establish a hydrogen nuclear magnetic quantitative method which can rapidly determine the contents of linalool and protocatechuate in Homalomena occulta. Methods: Using dimethyl sulfoxide-d₆(DMSO-d₆)as the test solvent and pyrazine -as the internal standard, ¹H-q NMR measurement was performed on a 400 MHz spectrometer. The quantitative resonance peaks of pyrazine, Linalool and protocatechuate were δ 8.66, δ 5.90 and δ 8.60, respectively. Results: The linear correlation coefficients of linalool and protocatechuate were good with r of 0.999 7 and 0.998 6, respectively. The RSDs of precision were 0.30% and 0.40%, and the recoveries were 98.8%-100.7%(RSD=3.7%) and 99.4%-100.6%(RSD=4.4%), respectively, which indicated that the established method was accurate, stable and feasible. And the whole detection process was completed in about three minutes. The contents of 2 components in 4 batches of Homalomena occulta were 0.328-0.398 mg·g^-1(linalool)and 0.559-0.630 mg·g^-1 (protocatechuate), respectively. Conclusion: The ¹H-q NMR method has the advantages of simple operation, fast analysis speed and specificity, and can be used for the simultaneous determination of two active components in Homalomena occulta, providing a scientific basis for the overall quality evaluation and quality control of Homalomena occulta.

Objective: The early warning of new psychoactive substance iso-propoxate (homologue of etomidate) was carried out through sewage drug monitoring technology, and quantitative research was conducted to provide technical support for management and law enforcement. Methods: The abnormal results were obtained by quantitative analysis of multiple reaction monitoring (MRM) mode using online solid phase extraction-ultra high performance liquid chromatography tandem mass spectrometry (SPE-UPLC-MS/MS), and the basic information of abnormal substance was determined by precursor ion scanning mode. The qualitative determination was carried out using parallel reaction monitoring (PRM) mode by ultra high performance liquid chromatography quadrupole-orbitrap high resolution mass spectrometry (UPLC-Q exactive HRMS) after the directional synthesis of the compound. Quantitative study was then conducted by commercial reference products. The correlation analysis was carried out by SPSS 27. Results: The abnormal substance was determined to be iso-propoxate. The optimized MRM parameters could be distinguished from its isomers by specific ion abundance ratio. The correlation analysis of quantitative results showed that it was similar to etomidate and ketamine abuse areas. Conclusion: In the process of sewage drug monitoring, it is necessary to pay attention to the proportional relationship between prototype and metabolite. It is important to summarize the characteristic fragments and characteristic neutral loss information of specific types of compounds, carry out screening non-targeted detection for key samples in key areas in the monitoring process, and analyze potential substances using high-resolution mass spectrometry and other methods, so as to fully leveraging the early warning role of sewage drug monitoring.

Objective: To compare the regulatory effects of authentic and counterfeit Arnebiae Radix on intestinal flora in mice based on metagenomic sequencing. Methods: Firstly, 24 clean grade female BLAB/C mice were randomly divided into 3 groups:blank control group, A1 (Arnebia euchroma) group and A2 (Arnebiae Radix whose origins were not included in Chinese Pharmacopoeia) group. After gavage arrived at the specified time, colon contents (feces), ileal contents (feces) and small intestinal contents (feces, except ileal parts) were extracted for intestinal flora analysis. Genomic DNA was extracted and amplified by PCR from the extracted mouse intestinal contents. The PCR products were mixed and purified. Then library was constructed and sequenced. After quality control of sequencing data and removal of chimera sequence, the final effective data was obtained. Operational taxonomic unit(OTU) clustering and species annotation were performed on the obtained valid data, and sample diversity analysis was conducted. Results: In this study, both A1 (Arnebia euchroma) group and A2 (Arnebiae Radix non-pharmacopoeia) group reduced the diversity of mice colon microbiota. At the phylum level, group A1 significantly increased the abundance of Firmicutes in the small intestine and ileum, and groups A1 and A2 significantly increased the relative abundance of Bacteroidetes in the colon. At the genus level, group A1 significantly increased the relative abundance of Lactobacillus in the small intestine of mice, and group A2 significantly increased the relative abundance of Lactobacillus in the ileum of mice. Group A1 increased the relative abundance of Lactobacillus, one of the dominant bacteria in the colon of mice, and group A2 increased the relative abundance of Bacteroides. Alistipes mainly existed in the colon, A2 group significantly reduced the relative abundance of Alistipes in the colon of mice, while Alistipes in the A1 group was cultivated. Conclusion: According to the results of the regulation effect of intestinal flora, the intestinal flora regulation effect in the two experimental groups of the authentic Arnebiae Radix and its confusion products from markets are not consistent, the results of this study can provide a theoretical basis for further exploring the mechanism of Arnebiae Radix.

Objective: To develop an HPLC method for the determination of related substances and assay of levamisole hydrochloride tablets. Methods: The chromatographic separation of related substances was performed on a Thermo Hypsil C₁₈ column(100 mm×4.6 mm, 3 μm). A gradient elution was applied with a mobile phase composed of 0.75% monobasic ammonium phosphate solution(adjusted at pH 6.5 with diiopropylamine) and acetonitrile. The assay analytical column was packed with IntertSustain C₁₈ column(150 mm×4.6 mm, 5 μm). The mobile phase was 70 volumes of 0.75% monobasic ammonium phosphate solution(adjusted at pH 7.0 with diiopropylamine) with 30 volumes of acetonitrile. The wavelength detection was set at 215 nm, the injection volumn was 10 μL, and the flow rate was 1.0 mL·min^-1. Results: Levamisole and its impurities were separated well by related substances HPLC method above. Impurities A, B, C, D, E showed the good linearities in the concentration ranges of 10.87-25.37 μg·mL^-1, 11.62-27.10 μg·mL^-1, 12.38-28.90 μg·mL^-1, 30.89-72.07 μg·mL^-1, 12.41-28.95 μg·mL^-1 (r＞0.999). The average correction factors of impurities A, B, C, D, E determined by three columns and liquid chromatographies were 1.6, 1.4, 2.6, 1.2, 2.4, respectively, the recovery rates(n=9) were 98.1%, 99.0%, 98.6%, 100.1%, 99.9% and the RSDs were 0.63%, 0.79%, 0.92%, 0.96%, 0.33%. The separation of levamisole and impurity C was good by assay HPLC method. Levamisole showed the good linearity in the concentration range of 0.10 14-0.304 3 mg·mL^-1 (r=0.999 9). The average recovery rate(n=9) of levamisole was 100.2%, RSD=0.52%. Ten batches levamisole hydrochloride tablets from five domestic pharmaceutical enterprises were determined by the above related substances and assay HPLC method, the total impurities mass were 0.05%-0.62%, and the assays were 99.5%-104.3%. Conclusion: The established method is high selection and accurate. It is suitable applied for determination of related substances and assay of levamiole hydrochloride tablets.