Abstract: Objective: To determine the content of danshensu and protocatechuic aldehyde in Salvia miltiorrhiza from five places. Methods: High performance liquid chromatography, Kromasil analytical column (250 mmx46mm, 5μm; mobile phase: methanol-1Og / L glacial acetic acid 20:80; UV detection wavelength: 280nm; flow rate: 1.0ml / min. Results: Danshensu 4.2-33.6μg / ml, protocatechualdehyde has a good linear relationship in the range of O.36-1.80μg / ml, the minimum detection limits of danshensu and protocatechualdehyde are 10.5 ng and 0.6 ng, respectively, and the recovery rates are 99.36% (RSD 1.26%) and 97.54% (RSD O.81%). Conclusion: The method is simple, sensitive and accurate, and the sample processing is simple and practical. It has practical value in the evaluation of the quality of Salvia miltiorrhiza. Salvia miltiorrhiza, its danshensu and protocatechuic aldehyde content have a certain difference. This article is derived from the determination of danshensu and protocatechuic aldehyde in Salvia miltiorrhizae from five different origins by high-performance liquid chromatography. Sleeping Mask,Slumber Mask,Shut-Eye Mask,Somnus Mask,Repose Mask Guangdong Yueji Cosmetics Co., Ltd , https://www.mask-family.com
Keywords: Salvia miltiorrhizae; Salvia miltiorrhizae; Protocatechuic aldehyde; High performance liquid chromatography method Salvia miltiorrhizae is the dried root and rhizome of Salvia miltiorrhiza Bge, which has the functions of removing blood stasis and pain, promoting blood circulation, clearing the mind and removing troubles . Modern pharmacological studies of traditional Chinese medicine have shown that Salvia miltiorrhiza can increase coronary blood flow, lower blood fat, inhibit thrombosis or anti-thrombosis, and improve microcirculation. Its main active ingredients are water-soluble phenolic acids (danshensu and protocatechuic aldehyde) and fat-soluble diterpene quinones (tanshinone â…¡ A). Salvia miltiorrhiza is used in traditional and modern medicine, mainly based on various preparations based on its water-soluble ingredients. Therefore, for the needs of clinical and scientific research, the author used high-performance liquid chromatography to determine the water-soluble components of Danshensu and protocatechuic aldehyde in Salvia miltiorrhiza, and compared the contents of danshensu and protocatechuic aldehyde in 5 kinds of Salvia miltiorrhiza.
1 Materials 1.1 Instruments High-performance liquid chromatography system (7725 manual injector, 600 pumps, 996 diode array detector, M data processing system) produced by Waters, USA, Digital 586 computer, TGL · 16G desktop high-speed centrifugation Machine, Sz-93 automatic double pure water distiller, TN-100 B type tray torque balance, GB-204 type electronic balance.
1.2 Salvia miltiorrhiza Bunge from Shaanxi Province; Luoyang Salvia miltiorrhiza from Henan Province; Pucheng Salvia miltiorrhiza from Shaanxi Province; Lu's family of Salvia miltiorrhiza, wild Salvia miltiorrhiza It was purchased from the Chinese Medicine Operation Department of Sanmenxia Medicine Purchasing and Supply Station in Henan Province. Salvia miltiorrhiza in the above five kinds of production areas was identified as the dried rhizome of Salvia mihiorrhiza Bge by the deputy chief pharmacist of the Institute of Pharmacy, the Fourth Military Medical University of the Chinese People's Liberation Army.
1.3 Reagent methanol, purchased from Tianjin Chemical Reagent Co., Ltd., alcohol analysis; glacial acetic acid, purchased from Shaanxi Xi'an Chemical Reagent Factory, alcohol analysis; Danshensu sodium standard: purchased from China National Institute for Pharmaceutical and Biological Products, No. 855- 200102; Protocatechualdehyde standard product: purchased from China National Institute for the Control of Pharmaceutical and Biological Products, No. 0810-9803; the water is homemade double distilled water.
2 Methods and results 2.1 Chromatographic conditions Column: KF series HPLC column of Dalian Institute of Physical Chemistry, Chinese Academy of Sciences. Kromasil Cl8: 250 mm x4.6 mm, 5 μm, NO: Cl8B 021110; mobile phase: methanol-10 g / L glacial acetic acid (20:80); flow rate: 1.0 ml / min; detection wavelength: 280 nm; detection sensitivity: 0.001AUFS ; Column temperature: 18 ℃; Resolution: 1O.34; The number of theoretical plates (calculated with Danshensu) is 6390.
2.2 Linear range Precisely weigh the appropriate amount of danshensu and protocatechualdehyde reference substance, and prepare the reference substance mother liquor with concentrations of 0.21 mg / ml and 0.12 mg / ml with 10 g / L glacial acetic acid, respectively. Pipette 200, 400, 800, 1000, 1200, and 1600 μl of Danshensu reference mother liquor with a micro sampler; 3O, 6O, 90, 105, 120, 150 μl of protocatechualdehyde reference mother liquor, add 10 ml of brown in the corresponding order Add 10 g / L glacial acetic acid to the graduation mark in the volumetric flask, and prepare 6 mixed reference solutions of different concentrations. Aspirate 5μl of the sample with a micro sampler, and call the peak area the area formed by the baseline. A = × σ × h = 2.507σh = 1.064 Wh / 2h> The peak area is the ordinate, the reference sample injection volume (ng) is the abscissa, and linear regression is performed (n = 6). The regression equation of danshensu and protocatechuic aldehyde is: y = -86.75 + 158.54x, r = 0.9997; Y = 16.45 + 1751.28x, r = 0.9990. The results showed that Danshensu exhibited a good linear relationship in the range of 4.2-33.6μg / ml and protocatechualdehyde in the range of 0.36-1.80μg / ml.
2.3 Accuracy test The mixed reference solution with the concentration of danshensu precisely 21.00 μg / ml and the concentration of protocatechuic aldehyde 1.26 μg / ml was re-injected 5 times. The peak area integral value of danshensu and protocatechuic aldehyde RSD They were 0.37% and 0.28% (n = 5).
2.4 Repeatability test Take the decoction of the medicinal material sample (Wild Salvia miltiorrhiza) from the same place, prepare 5 parts according to the sample solution pretreatment method, and inject 5μl respectively. The results of the peak area integral values ​​RSD of danshensu and protocatechualdehyde are respectively 0.38% and 0.33%.
2.5 Stability test Take the test solution (Salvia miltiorrhiza of the family Lu) from the same place of origin at 0, 2, 4, 6, and 8h, and record the peak areas of danshensu and protocatechualdehyde. The peak areas RSD were 1.17, respectively. % And 0.49%. It indicates that the test solution is stable.
2.6 Determination of the minimum detection limit Take a mixed reference solution, dilute with l0g ​​/ L glacial acetic acid to make a series of gradient concentration solutions from high to low, inject samples under the above chromatographic conditions, and take a peak height of 3 times the noise The injection volume at the time is the lowest detection concentration, and the minimum detection limit is obtained from the injection volume of 20 μl. Results of Danshensu and protocatechu
Determination of Danshensu and protocatechuic aldehyde in Salvia miltiorrhiza from 5 different regions by HPLC