1.Establishment of Content Determination Method of Zhihuoding Granules Based on Anti-inflammatory Activity and Fingerprint
Xin ZENG ; Guangzhi SHAN ; Wenling SU ; YIDERESI·MOLA ; Yue WANG ; Jianguo XING
Herald of Medicine 2025;44(7):1028-1034
Objective To establish the content determination method of Zhihuoding granules(ZHDG)by the anti-inflammatory active ingredients and fingerprint.Methods The inflammatory model of BEAS-2B cells was established by LPS(10 μg·mL-1)stimulation for 24 h,and the level of TNF-α in the supernatant was detected by enzyme-linked immunosorbent assay(ELISA).Using the reverse high-performance liquid chromatography(RP-HPLC)of 7 batch ZHDG fingerprint;HPLC was used to determine the content of geniposide and baicalin in granules.Results Compared with LPS Group,ZHDG,prim-O-glucosylcimifugin,5-O-methylvisammioside,baicalin,geniposide,and total polysaccharide extract all significantly decreased TNF-αlevels in BEAS-2B cells induced by LPS(P<0.05).Fingerprint spectra of 7 batches of ZHDG were established,with a similarity of 1.000 to the reference.A total of 15 peaks were calibrated,and 7 chromatographic peaks were identified.Gardenin and baicalin were selected as the index components of ZHDG for evaluation.The established method was rapid and accurate.Conclusion The method for the determination of baicalin and geniposide in ZHDG are exclusive,accurate,and effective,providing reference for the quality control and clinical application of ZHDG.
2.Establishment of Content Determination Method of Zhihuoding Granules Based on Anti-inflammatory Activity and Fingerprint
Xin ZENG ; Guangzhi SHAN ; Wenling SU ; YIDERESI·MOLA ; Yue WANG ; Jianguo XING
Herald of Medicine 2025;44(7):1028-1034
Objective To establish the content determination method of Zhihuoding granules(ZHDG)by the anti-inflammatory active ingredients and fingerprint.Methods The inflammatory model of BEAS-2B cells was established by LPS(10 μg·mL-1)stimulation for 24 h,and the level of TNF-α in the supernatant was detected by enzyme-linked immunosorbent assay(ELISA).Using the reverse high-performance liquid chromatography(RP-HPLC)of 7 batch ZHDG fingerprint;HPLC was used to determine the content of geniposide and baicalin in granules.Results Compared with LPS Group,ZHDG,prim-O-glucosylcimifugin,5-O-methylvisammioside,baicalin,geniposide,and total polysaccharide extract all significantly decreased TNF-αlevels in BEAS-2B cells induced by LPS(P<0.05).Fingerprint spectra of 7 batches of ZHDG were established,with a similarity of 1.000 to the reference.A total of 15 peaks were calibrated,and 7 chromatographic peaks were identified.Gardenin and baicalin were selected as the index components of ZHDG for evaluation.The established method was rapid and accurate.Conclusion The method for the determination of baicalin and geniposide in ZHDG are exclusive,accurate,and effective,providing reference for the quality control and clinical application of ZHDG.
3.A comparative study of NQAD and ELSD for the determination of peimine and peiminine in huangshi xiangsheng pills
Huiyi LIU ; Limin ZUO ; Xiaodan QIU ; Xin GUO ; Xiaofang LIAN ; Qingying JIA ; Yongsheng GU ; Xuanlin LIU ; Guangzhi SHAN
Drug Standards of China 2024;25(6):529-535
Objective:The study aimed to quantify the contents of peimine and peiminine in huangshi xiangsheng pills using a nano quantity analyte detector(NQAD)and compare the results with the evaporative light scattering detector(ELSD).Methods:Analytical separation was conducted utilizing a CAPCELL PAK MG Ⅲ C18 column(4.6 mm ×250 mm,5 μmi)as the stationary phase.The mobile phase consisted of a mixture of acetonitrile-water-diethylamine(65∶35∶0.03),with a flow rate set at 1.0 mL·min-1 and the column maintained at a temperature of 25℃.Results:NQAD analysis revealed a robust linear correlation between the concentration and the peak area for peimine within a concentration range of 2.450 to 1.470 μg·mL-1,achieving a correlation coefficient of 0.999 3.The limit of detection(LOD)and recovery rate for peimine were determined to be 14.70 ng and 99.50%(n=9),respectively.Similarly,a linear relationship was observed for peiminine between the concentration and the peak area,spanning a range of 3.657 to 1.170 μg·mL-1,with a correlation coefficient of 0.999 1.The LOD and recovery rate for peiminine were 21.94 ng and 100.5%(n=9),respectively.In contrast,ELSD yielded LOD of 24.50 ng for peimine and 36.57 ng for peiminine,with linear range of double logarithmic fitting was 4.900 to 490.0 μg·mL-1 and 7.314 to 585.1 μg·mL-1,respectively.Conclusion:NQAD outperforms ELSD in the quantification of peimine and peiminine,offering superior accuracy,precision,and sensitivity,along with an extended dynamic linear range.By enabling direct content calculation via linear regression,it streamlines the process,bypassing the complex double logarithmic calculations.This approach not only boosts the efficiency of data handling but also substantially simplifies the computational steps,addressing the detection and analysis needs for peimine and peiminine in huangshi xiangsheng pills.
4.A comparative study of NQAD and ELSD for the determination of peimine and peiminine in huangshi xiangsheng pills
Huiyi LIU ; Limin ZUO ; Xiaodan QIU ; Xin GUO ; Xiaofang LIAN ; Qingying JIA ; Yongsheng GU ; Xuanlin LIU ; Guangzhi SHAN
Drug Standards of China 2024;25(6):529-535
Objective:The study aimed to quantify the contents of peimine and peiminine in huangshi xiangsheng pills using a nano quantity analyte detector(NQAD)and compare the results with the evaporative light scattering detector(ELSD).Methods:Analytical separation was conducted utilizing a CAPCELL PAK MG Ⅲ C18 column(4.6 mm ×250 mm,5 μmi)as the stationary phase.The mobile phase consisted of a mixture of acetonitrile-water-diethylamine(65∶35∶0.03),with a flow rate set at 1.0 mL·min-1 and the column maintained at a temperature of 25℃.Results:NQAD analysis revealed a robust linear correlation between the concentration and the peak area for peimine within a concentration range of 2.450 to 1.470 μg·mL-1,achieving a correlation coefficient of 0.999 3.The limit of detection(LOD)and recovery rate for peimine were determined to be 14.70 ng and 99.50%(n=9),respectively.Similarly,a linear relationship was observed for peiminine between the concentration and the peak area,spanning a range of 3.657 to 1.170 μg·mL-1,with a correlation coefficient of 0.999 1.The LOD and recovery rate for peiminine were 21.94 ng and 100.5%(n=9),respectively.In contrast,ELSD yielded LOD of 24.50 ng for peimine and 36.57 ng for peiminine,with linear range of double logarithmic fitting was 4.900 to 490.0 μg·mL-1 and 7.314 to 585.1 μg·mL-1,respectively.Conclusion:NQAD outperforms ELSD in the quantification of peimine and peiminine,offering superior accuracy,precision,and sensitivity,along with an extended dynamic linear range.By enabling direct content calculation via linear regression,it streamlines the process,bypassing the complex double logarithmic calculations.This approach not only boosts the efficiency of data handling but also substantially simplifies the computational steps,addressing the detection and analysis needs for peimine and peiminine in huangshi xiangsheng pills.
5.Gene clone, eukaryotic expression vector and tissue expression profile analysis of porcine BST-2 gene
Ning KONG ; Yongguang WU ; Qiong MENG ; Zhongze WANG ; Wu TONG ; Hao ZHENG ; Guoxin LI ; Tongling SHAN ; Enmin ZHOU ; Guangzhi TONG
Chinese Journal of Veterinary Science 2017;37(8):1594-1599,1640
In order to study the biological function of pig BST-2 gene,the BST-2 gene was amplified with specific primers from porcine kidney tissue,and molecular characterization of BST-2 nuclectide and amino acid sequence were analyzed with bioinformatics tools and online server.Then the prokaryotic expression and tissue expression profile analysis was carried out.The results showed that the full length of pig BST-2 gene was 851 bp and contained 23 bp of 5'-UTR,294 bp of 3'-UTR and 534 bp of CDS and the gene encoded 177 aa.Amino acid sequence analysis of pig BST-2 protein showed 46.1% identity with gorilla gorilla,41.7% with cricetulus griseus,39.5% with mus musculus,35.4% with equus asinus,42.0% with felis catus,40.5% with bos mutus,44.4% with macaca mulatta,38.7% with ovis aries and 46.8% with homo sapiens.BST-2 protein contained 2 transmembrane structure (27-49 aa and 154-176 aa),2 glycosylation sites and 14 potential phosphorylation sites including ATM,CK Ⅱ,PKA,PKC binding sites.The pig BST-2 protein was expressed in Vero cells after translated the recombinant plasmid FLAG-BST-2.Semiquantitative PCR results showed that BST-2 gene was expressed in all the tissues,especially in lymph nodes,thymus,tonsils,spleen,large intestine and small intestine.This study provide a foundation for further understanding the antiviral mechanism of pig BST-2 protein.
6.Content Determination of Urea in Urea [13C] Capsules by High Performance Cation-exchange Chromatogra-phy
Guixia LIU ; Jing YAO ; Limin ZUO ; Panpan LI ; Guangzhi SHAN ; Wei JIANG
China Pharmacy 2017;28(9):1236-1238
OBJECTIVE:To determine the content of urea in Urea [13C] capsules by high performance cation-exchange chroma-tography (HPCEC). METHODS:The determination was performed on Zorbax 300 SCX column with mobile phases consisting of acetonitrile-0.1% phosphoric acid (20:80,V/V) at the flow rate of 1.0 mL/min. The detection wavelength was set at 200 nm and column temperature was 35 ℃. The sample size was 20 μL. RESULTS:The linear range of urea was 0.0039-1.0030 mg/ml(r=0.9997). The limit of quantitation was 3.918 μg/mL and the limit of detection was 0.975 μg/mL. RSDs of precision,stability and repetitive test were all lower than 2.0%. The recovery ranged 99.3%-101.0%(RSD=0.67%,n=9). CONCLUSIONS:The meth-od is simple,rapid,sensitive and suitable for the content determination of urea in Urea [13C] capsules.
7.Contents Determination of Citric Acid in Fentanyl Citrate Raw Materials and Its Injection by Ion Chroma-tography
Xun MA ; Panpan LI ; Guixia LIU ; Yanping ZONG ; Wei SUN ; Guangzhi SHAN ; Hua CHEN ; Nan NAN
China Pharmacy 2017;28(18):2536-2538
OBJECTIVE:To determine the contents of citric acid in fentanyl citrate raw materials and its injection by ion chro-matography. METHODS:The determination was performed on Thermo Dionex IonPacTM AS11-HC column with mobile phase con-sisted of potassium hydroxide (gradient elution) at the flow rate of 1.0 mL/min. The column temperature was 35 ℃,and sample size was 20 μL. The detector was suppressed conductivity detector. RESULTS:The linear range of citric acid were 0.1157-74.05 μg/mL(r=0.9995). The limit of quantitation was 0.1150 μg/mL,and the limit of detection was 0.0400 μg/mL;RSDs of preci-sion,stability and reproducibility tests were all lower than 2.0%;the average recoveries were 99.6%-101.5%(RSD=0.68%,n=9). CONCLUSIONS:The method is environmentally-friendly and simple with good accuracy and precision,and suitable for the contents determination of citric acid in fentanyl citrate raw materials and injection.
8.Shan Guangzhi's Experience for the Treatment of Bone Metastasis of Prostate Cancer
Journal of Zhejiang Chinese Medical University 2016;40(2):131-133
Objective] To sum up professor Shan Guangzhi's clinical experience for the treatment of bone metastasis of prostate cancer. [Method] From following professor Shan Guangzhi in the clinical practice and further studying his medical cases,summarizes his experience in the treatment of prostate cancer bone metastases,and sums up his academic thoughts for the treatment of the disease.[Result] Professor Shan Guangzhi thinks that deficiency of the kidney is the basic pathogenesis of bone metastases of prostate cancer,at the same time accompanied by spleen deficiency and qi-movement disturbance. To prostate cancer patients with bone metastases, pain is the most common symptom,the reason for it is that stagnation leading to pain and poor nutrition lead to pain. The treatment should focus on two aspects,both the inner treatment and the external treatment; the internal treatment regards invigorating Qi and tonifying the kidney as the basis,also pays attention to regulate spleen and stomach,dredges the body down;the external treatment regards promoting blood circulation to remove meridian obstruction as the rule,it can effectively relieve patients' pain,and cooperates with diet,as a whole to improve the quality of survival in patients. [Conclusion]Professor Shan Guangzhi's experience in the treatment of bone metastasis of prostate cancer has exact curative effect,worth learning and promotion.
9.Content Determination of Acetic Acid in Octreotide Acetate for Injection by IEC
Jinghua LI ; Guixia LIU ; Panpan LI ; Zhiliang WANG ; Jing YAO ; Zhuorong LI ; Guangzhi SHAN
China Pharmacy 2016;27(27):3867-3869
OBJECTIVE:To determine the content of acetic acid in Octreotide acetate for injection by IEC,and provide reference for the improvement of pharmacopoeia standards. METHODS:The column was Rezex ROA-Organic Acid H+ with mobile phase of 0.002 5 mol/L sulfuric acid at a flow rate of 0.5 ml/min,the detection wavelength was 210 nm,column temperature was 45℃,and in-jection volume was 100 μl. RESULTS:The linear range of acetic acid was 0.394 4 μg/ml-78.89 μg/ml(r=0.999 9);RSDs of preci-sion,stability and reproducibility tests were all lower than 2%;the limit of quantification was 197.2 ng/ml,and limit of detection was 78.89 ng/ml;recovery was 104.71%-109.78%(RSD=1.34%,n=9). CONCLUSIONS:The method is environmental and simple with good accuracy and precision,and suitable for the content determination of acetic acid in Octreotide acetate for injection.
10.Determination of 2460 A in Trichoderma Hazianum Fermentation Liquor by On-line Two-dimensional Liquid Chromatographic Method
Guangzhi SHAN ; Jie ZHOU ; Limin ZUO ; Wei JIANG ; Guixia LIU ; Yang ZHANG ; Yuan LI ; Rong JIANG
Chinese Journal of Analytical Chemistry 2014;(12):1828-1832
An on-line two dimensional liquid chromatographic (2D-LC) method was established by using an ultimate dual gradient liquid chromatography, three chromatographic columns and valve-switching technology to detect 2460A in trichoderma hazianum fermentation liquor. MF C8 column (10 mm×4. 6 mm, 5. 0 μm) was used as purification column and MG C18 column (20 mm×4. 6 mm, 5. 0 μm) was used as enriching column. Methanol and water were used as mobile phase with a gradient elution at a flow rate of 2. 0 mL/min. The sample was separated on the Thermo Hypersil GOLD C18 column (250 mm×4. 6 mm, 5. 0μm) maintained at 40 ℃ using methanol and water. The flow rate was 1. 0 mL/min and 1. 0 mL sample was injected into the 2D-LC system. The detection wavelength was 424 nm. The whole analytical time was less than 60 min. The standard curve was linear over the 2460A concentration range of 0. 0025-10. 0 mg/L(r=0. 9981, n=8). The limit of detection was calculated to be 1 . 2μg/L ( S/N=3 ) and the limit of quantification was calculated to be 2 . 5 μg/L ( S/N=10 ) . The average recoveries varied from 88 . 0% to 104 . 4%.

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