高效液相色谱法测定复方荔枝草颗粒剂中高车前苷含量（Determination of homoplantaginin in Compound Salvia plebeia granules by HPLC）

高效液相色谱法测定复方荔枝草颗粒剂中高车前苷含量（Determination of homoplantaginin in Compound Salvia plebeia granules by HPLC） 高效液相色谱法测定复方荔枝草颗粒剂中高车前苷含量 (Determination of homoplantaginin in Compound Salvia plebeia granules by HPLC) High performance liquid chromatographic determination of the content of high glucoside in compound litchi granules Wu Zhenghong, Pei Yunping, Fang Yun, Chen Zhiyi High performance liquid chromatographic determination of the content of high glucoside in compound litchi granules Wu Zhenghong, Pei Yunping, Fang Yun, Chen Zhiyi Abstract Objective: to establish a HPLC method for the determination of the content of high purity compound in compound litchi granules. Methods: KF-C18 (250 mm * 4.6 mm, 10 m) for the analysis column, the mobile phase of 1 mol.L-1 phosphate buffer methanol water -pH 3.5 (40 / 55 / 5); the column temperature is 45 DEG C; the detection wavelength was 335 nm; the flow rate was 1 mL.min-1, the area external standard method. Results: the linear range was 3.12 * 10-3 to 0.20 g, and the average recovery was 96.96% ~ 101.5%, RSD < 0.69% (n=3). Conclusion: this method is simple, sensitive and reproducible. It can be used for the determination of the content and quality standard of high content of the granule. Keywords HPLC; high dose liquid chromatography (HPLC) HPLC; high resolution liquid chromatography (HPLC); Determination of Homoplantaginin In, Compound, Salvia, plebeia, Granules, by, HPLC Wu, Zhenghong, Pei, Yunping, Fang, Yun, and, Chen, Zhiyi (The, Clinical, Pharmacological, Department, of, the, Drum, Tower, Hospital,, Att, ached, to, the) Medical, College, of, Nanjing, University, Nanjing 210008) Abstract Objective:To establish a HPLC method for the determi nation of homoplantaginin in compound Salvia plebeia granules.Method s:Chromatographic conditions were as follows:analytical column of KF-C 18 (250 mm * 4.6 mm, 10 m), mobile phase of methanol-water-1.0 mol.L -1 pH phosphate buffer, 3.5 (40 / 55 / 5), column temperature at 45 C, D etective wavelength at 335 nm, 1 mL.min-1 of flow rate.The external st andard method was used. Results:The standard curve was linear in the range of 3.12 * 1 0-3 ~ 0.20 g, the average recovery was 96.96% ~ 101.5%, and RSD w as less than 0.69% (n=3). Conclusion:The method is simple, sensitive and, highly reproducibl e.The content of homoplantaginin in the granules was determined by the method An, D, quality, standard, of, the, granules, was, formulated. Key, words, homoplantaginin, compound, Salvia, plebeia, granules, HPLC Compound salvia plebeia granules is mainly composed of herha (Salvia plebeia R.Br.), plantain (Plantago asiatica L.) and Serissa serissoides June snow (DC.) composed of traditional Chinese medicine Druce, has the efficacy of detoxification, diuretic tonglin. The main herbs, litchi herb and plantain, have the effects of diuresis, clearing heat and detumescence (1, 2), and all contain flavonoid glycosides, high [3 [4]]. In order to control the quality of the preparation, the quantitative analysis of the content of the precursor was carried out. This method was accurate, sensitive and stable. 1 instrument and reagent SHIMADZU LC-10A high performance liquid chromatography, SPD-10A UV detector, C-R7A data processor. Homoplantaginin control (provide content above 98%, Shanghai Pharmaceutical Industry Research Institute); compound salvia plebeia granules (the production of Gulou Hospital Affiliated to Medical College of Nanjing University); methanol for HPLC pure phosphoric acid, potassium dihydrogen phosphate, were analytically pure. 2 experimental methods and results 2.1 chromatographic column: KF-C18 (4.6 mm * 250 mm, 10 m); UV detection wavelength: 335 nm; the mobile phase of 1 mol.L-1 phosphate buffer methanol water -pH 3.5 (potassium dihydrogen phosphate 68.05 g, the amount of water, dissolved, adding phosphoric acid 1.6 m L, and then dilute with water to 500 mL (40): 55: 5); column temperature: 45 C; flow rate: 1 mL.min -1; paper speed: 2 mm.min-1. 2.2 homoplantaginin standard curve precision weighing homoplantaginin control amount of goods, adding methanol dissolved, diluted solution of 0.2 mg.mL-1, and the mobile phase solution, preparation of concentration of 10, 5, 2.50, 1.25, 0.625, 0.312, 0.156 g.mL-1 of the reference solution, precise amount of 20. The L sample, determined according to the above conditions. The high peak concentration (X) was plotted by chromatographic peak area (Y), and the work curve was obtained and its regression equation was obtained: Y=33 082.50X-1 26.33 r=1.000 0 Good linear relation was found in the range of 0.156 to 10 g.mL-1. 2.3 samples from compound salvia plebeia granules crushed, accurately weighed about 1 g, a conical flask, adding 60% ethanol, 10 mL precision, weighing, the ultrasonic oscillator in the sound of 1.5 h, after cooling, and weighing, adding 60% ethanol to the original weight, filtering, discarding the initial filtrate, collecting filtrate continued and the precise amount of filtrate 1 mL in 25 m L volumetric flask, dilute with mobile phase to the scale, shake, sample analysis. In accordance with the external standard method, the content of the precursor was calculated by peak area, and the content of 3 batches of samples was 0.451, 0.512, 0.711, mg.g-1, RSD, 0.84%, 0.81%, 0.75% (n=3), respectively. The chromatogram is shown in figure 1. Figure 1 HPLC diagram A. high contrast extract B., compound litchi grass granules extract C. extract of compound litchi grass granules + tall plantain reference substance 1. tall plantain (16.6 min) 2.4 recovery test taking known homoplantaginin content of Compound Salvia plebeia granules crushed, accurately weighed about 1 g, a total of 3 copies in each were added 0.5, 1, 3 mL concentration of homoplantaginin 221.2 g.mL-1 standard solution (solvent for 60% ethanol), then adding 60% ethanol 9.5, 9, 7 mL, according to the determination of samples under operation, made of 3 different concentrations of the test solution, each concentration was repeated 3 times, calculating homoplantaginin content and recovery rate, the recovery rate is 96.96% ~ 101.5%, RSD < 0.69%. Homoplantaginin 2.5 precision test with high, medium and low 3 different concentrations of the standard solution, measured under operation according to the standard curve of 1 D and 1 Sunday between the peak area variability (n=5), the days of RSD < 3.3%, RSD < 4.7% day. 2.6 repetitive inspection from compound salvia plebeia granules crushed, accurately weighed about 1 g, according to the determination of samples under operation, preparing test solution 5, sample, each repeated 3 times to calculate the content of homoplantaginin in Compound Salvia plebeia granules, the average content is 0.507 mg.g-1, RSD 2.3% (n=15). 2.7 different extraction methods on the extraction rate of homoplantaginin from compound salvia plebeia granules crushed, take an appropriate amount, a total of 3, with 60% ethanol as solvent, respectively by Soxhlet extraction, ultrasonic extraction and reflux extraction method, 1.5 h, and then measured, and calculate the content of homoplantaginin in Compound Salvia plebeia granules the best effect, the results show that the extraction by ultrasonic method. 2.8 different solvents on the extraction rate of homoplantaginin from compound salvia plebeia granules crushed, accurately weighed about 1 g, a total of 7 copies, respectively with methanol, water and 50%, 60%, 65%, 70%, 95% ethanol as solvent ultrasonic extraction, and then press the sample under operation, preparing for the test solution, sample determination of homoplantaginin, calculate the content of Compound Salvia plebeia granules. The results showed that ultrasonic extraction with 60% ethanol as the best solvent. 3 discussion 3.1 in this paper, ultrasonic method is used to optimize the extraction process, and the extraction process is simplified. The cavitation, mechanical pulverization and agitation of ultrasonic vibration are beneficial to the penetration of chemical solvent into the interior of the extracted material, The active ingredient is accelerated into the solvent to promote the extraction of the active ingredient, thereby increasing the rate of the active ingredient. 3.2 blank samples made of extracts of litchi and plantain seeds were determined by this method. No peaks were detected at the corresponding positions in the high chromatogram chromatogram, which did not interfere with the results of hplc. Wu Zhenghong (Department of clinical pharmacology, Drum Tower Hospital, Medical College of Nanjing University, 210008) Pei Yunping (Department of clinical pharmacology, Drum Tower Hospital, Medical College of Nanjing University, 210008) Fang Yun (Department of clinical pharmacology, Drum Tower Hospital, Medical College of Nanjing University, 210008) Chen Zhiyi (Department of clinical pharmacology, Drum Tower Hospital, Medical College of Nanjing University, 210008) Reference 1 、 Institute of medicinal plant resources development, Chinese Academy of Medical Sciences, Institute of Materia Medica, Chinese Academy of Medical Sciences. Annals of traditional Chinese medicine. Second Edition (Fourth Edition). Beijing: People's Medical Publishing House, 1988.542 2, the Chinese Pharmacopoeia.1995. Part 1: 53 3, Jiang Yi, Ruth Qi, Zheng Minshi. Study on active ingredients of litchi grass. Pharmaceutical industry, 1987, 18 (8):3 49 4. Xiao Chonghou. Chinese medicine chemistry. First edition. Shanghai: Shanghai science and Technology Press, 1988.194 (this paper was received in August 26, 1999)