CN112684053B - A method for simultaneous determination of the content of 7 components in an anti-chronic heart failure drug compound - Google Patents

Abstract

The invention relates to a method for simultaneously measuring the contents of seven components in an anti-chronic heart failure medicine compound. The 7 components are gallic acid, hydroxysafflor yellow A, loganin, agarwood tetraol, psoralen, isopsoraside, and secretinoside, and the method is high performance liquid chromatography, The chromatographic conditions are as follows: a C ₁₈ chromatographic column, mobile phase acetonitrile: water, gradient elution; flow rate 0.2-1.2 mL·min ¹ , column temperature 20-40° C., detection wavelength 220-360 nm. The method can realize the simultaneous determination of the contents of 7 components in an anti-chronic heart failure drug compound. The formulation of the quality standard of anti-chronic heart failure drug compound provided the basis.

Description

A method for simultaneous determination of the content of 7 components in an anti-chronic heart failure drug compound

technical field

The invention relates to the field of quality control of traditional Chinese medicine compound prescriptions, in particular to a method for simultaneously measuring the contents of seven components in an anti-chronic heart failure medicine compound prescription.

Background technique

The anti-chronic heart failure drug compound, namely Bushen Huoxue ^Recipe , has been used in clinical treatment of chronic heart failure for many years. In dealing with the relationship between specimens, this prescription is based on tonic and seeks the essence, taking into account congestion, water and other standards; in dealing with the proportion of yin and yang, it balances yin and yang, mainly tonifies yang, and seeks yang in yin; in dealing with the relationship between the whole heart and kidney Emphasizes the intersection of the heart and the kidney, the heart originates from the kidney, and seeks to strengthen the heart by warming the kidney. The prescription is composed of 7 herbs: antler, safflower, epimedium, psoralea, privet japonica, dogwood and agarwood. Activating blood and clearing the meridians, the two medicines are the king medicine; Epimedium, Psoralea, Ligustrum lucidum, and Cornus help the antlers to calm the yin and yang of the kidney, and they are the minister medicine; Agarwood absorbs the qi and returns to the kidney to help the kidney medicine, and promotes qi and relieves pain. Helping safflower to promote blood circulation, and to make this formula tonic without stagnation, it is an adjuvant medicine; the combination of various medicines has the effect of tonifying the kidney and activating blood ^[1] . The anti-chronic heart failure drug compound can act on myocardial fibroblasts in a paracrine manner by activating the gp130/JAK2/STAT3 pathway to inhibit myocardial fibrosis ^[2] , and can also affect plasma RASS products ^[3,4] , which is considered to be a Risk factors for myocardial fibrosis.

Modern pharmacological studies have shown that antler polypeptide, the active ingredient of antlers, can mediate the differentiation of myocardial stem cells and repair damaged myocardium ^[5] ; hydroxysafflor yellow A is a compound with a monochalcone glycoside structure, which is the most effective pharmacological effect of safflower. The water-soluble part has obvious protective effect on vascular tissue cells, and has the functions of lowering blood pressure, dilating blood vessels, improving blood supply to organs, anti-coagulation, and anti-inflammatory ^[6,7] ; Psoralea has a variety of biological activities, and coumarin Epimedium is its main active ingredient, among which psoralen has a protective effect on the cardiovascular system ^[10] ; Epimedium is mainly composed of flavonoids, which have a great effect on fighting heart failure and protecting heart function. Its representative ingredients Icariin can increase the systolic function of the heart, reduce the load on the heart, and increase the cardiac output, thereby protecting the heart and blood vessels ^[8,9] ; Cornus is the main medicinal material for the treatment of diabetes, coronary heart disease and hypertension, and iridoids are The most abundant taxa in Cornus, Kang et al ^[11] showed that gallic acid increased NO content by up-regulating the phosphorylation level of endothelial nitric oxide synthase (eNOS), while gallic acid could inhibit angiotensin I converting enzyme ( ACE), significantly reduced blood pressure in spontaneously hypertensive rats, and was comparable to the pharmacological effect of captopril. Morroniside and loganin have a certain protective effect on the myocardium of diabetic mice ^[12] ; the active ingredients contained in Ligustrum lucidum can not only effectively regulate the human endocrine system in both directions, but also increase coronary blood flow, and have a negative effect on myocardial blood flow. It has an inhibitory effect on contractility, and has achieved good results in the treatment of cardiac insufficiency and arrhythmia ^[10,13] . Teluceoside ^[14] can significantly inhibit the apoptosis of glomerular mesangial cells and increase Bcl- 2/Bax and down-regulates the high expression of cleaved-caspase3 protein, thereby inhibiting premature and excessive apoptosis of glomerular mesangial cells, protecting cells, preventing renal failure and reducing proteinuria.

The research on the quality control of traditional Chinese medicine compounds is the key to the modernization and internationalization of traditional Chinese medicine. The patent document CN111175392A, published on 2020.05.19, discloses a quality control method for cough-relieving loquat syrup, and establishes an HPLC method for the content determination of amygdalin, mulberry B and rabathine. Patent document CN110780018A, published on 2020.02.11, discloses a quality control method for Jinggan Granules, and establishes a method for detecting the content of emodin and chrysophanol by HPLC. Patent document CN112098563A, published on 2020.12.18, discloses a method for detecting the chemical components of Bushen Huoxue Recipe. The traditional Chinese medicine components of Bushen Huoxue Recipe are Eucommia ulmoides, Bakuchichi, Achyranthes huai, Salvia miltiorrhiza, Wei Lingxian and papaya. Low back pain. Phase carrying out n-butanol extraction, concentrating the obtained water phase to obtain the liquid to be tested, and then using specific detection conditions to carry out UPLC-MS detection, the obtained test results are compared with the standard chromatogram and natural product database to obtain the chemical composition in the liquid to be tested , the advantage of the invention is that 93 kinds of effective chemical components in Bushen Huoxue Recipe can be qualitatively detected at the same time. But at present, there is no quality control study on the anti-chronic heart failure drug compound as described in this application, nor has the simultaneous determination of the seven components in the anti-chronic heart failure drug compound gallic acid, hydroxysafflor yellow A, loganin, Method for the content of agarwood tetraol, psoralen, isopsoraside, and tegretinoside.

SUMMARY OF THE INVENTION

The purpose of the present invention is to aim at the deficiencies in the prior art, and provide a method for simultaneously measuring the content of 7 components in the compound prescription of anti-chronic heart failure medicine.

Another object of the present invention is to provide a quality control method for an anti-chronic heart failure drug compound.

For realizing above-mentioned first purpose, the technical scheme that the present invention takes is:

A high-performance liquid chromatography method for simultaneously determining the content of 7 components in an anti-chronic heart failure drug compound, wherein the 7 components are gallic acid, hydroxysafflor yellow A, loganin, agarwood tetraol, and psoralen , Isopsoraside and Ateprivaside, the chromatographic conditions of the high performance liquid chromatography method are: adopt C ₁₈ chromatographic column, mobile phase acetonitrile-0.1% phosphoric acid water, elution gradient is 0 ~ 10min, 5% acetonitrile ; 10～15min, 5%～10% acetonitrile; 15～35min, 10%～18% acetonitrile; 35～40min, 18%～25% acetonitrile; 40～60min, 25%～35% acetonitrile; flow rate 0.8～1.2mL ·min ^-1 , column temperature 28～32℃, detection wavelength 220～360nm.

As a preferred embodiment of the present invention, the compound traditional Chinese medicine composition of the anti-chronic heart failure drug is: 10-20 parts by weight of antlers, 6-12 parts by weight of safflower, 15-25 parts by weight of psoralen, 25 parts by weight of Epimedium -35 parts by weight, 10-20 parts by weight of dogwood meat, 25-35 parts by weight of Ligustrum lucidum, and 4-8 parts by weight of agarwood.

As another preferred embodiment of the present invention, the method for preparing the test solution of the high performance liquid chromatography method is as follows: accurately weigh each medicinal material, add 8-15 times of water to decoct for 60-90 min, filter, and filter the filtrate. Concentrate under reduced pressure, transfer to a volumetric flask, make up to volume, shake well, filter through a microporous membrane, and take the subsequent filtrate.

As another preferred embodiment of the present invention, the preparation method of the mixed reference solution of the high performance liquid chromatography method is as follows: accurately weighing gallic acid, hydroxysafflor yellow A, loganin, agarwood tetraol, tonic For the reference substances of bakuchioside, isopsoraside, and tegretinoside, methanol was added to prepare mixed reference substance solutions with concentrations of 2.71, 8.61, 7.61, 643, 17.42, 11.88, and 45.00 μg/mL, respectively.

As another preferred embodiment of the present invention, the injection volume is 1-12 μL.

More preferably, the injection volume is 10 μL.

As another preferred embodiment of the present invention, the flow rate is 1.0 mL·min ^-1 .

As another preferred embodiment of the present invention, the column temperature is 30°C.

As another preferred embodiment of the present invention, the detection wavelength is 220-240 nm.

For realizing the above-mentioned second purpose, the technical scheme adopted by the present invention is:

A quality control method for an anti-chronic heart failure drug compound, comprising the steps of any one of the high performance liquid chromatography methods described above.

The advantages of the present invention are:

1. The present invention establishes the contents of seven components gallic acid, hydroxysafflor yellow A, loganin, agarwood tetraol, psoralen, isopsoraside, and privatide in the anti-chronic heart failure medicine compound. The method of determination, and the methodological verification of system adaptability, specificity, linearity, precision, repeatability and stability of the detection method of the seven indicators to determine the feasibility of the detection method of the seven components. The method has a good linear relationship, high precision, strong stability, good repeatability, high sample recovery rate, and is simple and easy to implement.

2. The parameters involved in the detection method of the present invention are obtained on the basis of the inventor's research experience, and an excellent detection effect is achieved under this detection condition.

3. The present invention provides a basis for the formulation of the quality standard of the anti-chronic heart failure drug compound.

Description of drawings

Figure 1: Mixed reference HPLC chart (A) and test product HPLC chart (B). 1. Gallic acid; 2. Hydroxysafflor yellow A; 3. Loganin; 4. Agarwood tetraol;

Figure 2: Chromatograms under the same chromatographic conditions with different mobile phases. A: 0.1% formic acid-(B) methanol system; B: 0.1% formic acid-acetonitrile system.

Figure 3: Chromatograms at different elution gradients. A: The elution gradient is 0～8min, 5% acetonitrile; 8～60min, 5%～50% acetonitrile; The recording time is 0～60min; B: The elution procedure is acetonitrile-0.1% phosphoric acid water, elution Gradient: 0～5min, 5% acetonitrile; 5～15min, 5%～15% acetonitrile; 15～60min, 15%～45% acetonitrile; recording time: 0～60min; C: The elution program is acetonitrile- 0.1% phosphoric acid water, elution gradient: 0～5min, 5% acetonitrile; 5～15min, 5%～15% acetonitrile; 15～41min, 15%～27% acetonitrile; 41～43min, 27%～30% acetonitrile; 43～60min, 30%～40% acetonitrile; 60～61min, 40%～5% acetonitrile; recording time is 0～60min; D: The elution program is acetonitrile-0.1% phosphoric acid water, the elution gradient is 0 ～10min, 5% acetonitrile; 10～15min, 5%～10% acetonitrile; 15～35min, 10%～18% acetonitrile; 35～40min, 18%～25% acetonitrile; 40～60min, 25%～35% acetonitrile ; The recording time is 0～60min.

Figure 4: Chromatograms of the anti-chronic heart failure drug compound at different wavelengths. A. 211 nm; B. 230 nm; C. 254 nm; D. 265 nm; E. 280 nm; F. 300 nm; G. 330 nm.

Detailed ways

The specific embodiments provided by the present invention will be described in detail below with reference to the accompanying drawings.

Example 1

1 Materials and methods

1.1 Materials and Instruments

Agilent 1100series high performance liquid chromatograph (Agilent DAD detector, Agilent Technologies, USA); BSA124S-CW electronic analytical balance (1/10,000), BT25S electronic analytical balance (1/100,000, Sartorius Scientific Instruments) (Beijing) Co., Ltd.). Hydroxysafflor yellow A (batch number AF9081601, content 98%), loganin (batch number AF7110703, content 98%), psoralen (batch number AF9102107, content 98%), isopsorain (batch number AF9102107, content 98%) AF9102108, content 98%) were purchased from Shanghai Hongyong Biotechnology Co., Ltd.; Agarwood tetraol (batch number DST190801-042, content 99%) and gallic acid (batch number DST190715-008, content 99%) were purchased in Chengdu Lemeitian Pharmaceutical Technology Co., Ltd.; special privet glycosides (batch number 2018011201, content 95%) reference substance were purchased from Chengdu Pulis Biotechnology Co., Ltd. Deer antler, safflower, psoralen, epimedium, dogwood, privet lucidum, and agarwood decoction pieces were all purchased from Shanghai Kangqiao Traditional Chinese Medicine Pieces Co., Ltd., and were identified as genuine by researcher Wang Changhong of Shanghai Standardization Research Center of Traditional Chinese Medicine. Acetonitrile (Fisher Company, USA); formic acid (batch number: G0960200, content 98%) were purchased from Shanghai Anpu Scientific Instrument Co., Ltd.; other reagents were analytically pure; water was ultrapure water.

1.2 Test methods and results

1.2.1 Chromatographic conditions

The chromatographic column is ZORBAX SB-Aq (4.6mm×250mm, 5μm), the mobile phase is acetonitrile-0.1% phosphoric acid water, the elution gradient is 0～10min, 5% acetonitrile; 10～15min, 5%～10% acetonitrile; 15 ～35min, 10%～18% acetonitrile; 35～40min, 18%～25% acetonitrile; 40～60min, 25%～35% acetonitrile; flow rate 1.0mL·min ^-1 , column temperature 30℃, detection wavelength 230nm, 10 μL was injected, and the HPLC chromatogram of the mixed reference solution and the test solution was shown in Figure 1.

1.2.2 Preparation of mixed reference solution

Accurately weigh the appropriate amount of gallic acid, hydroxysafflor yellow A, loganin, agarwood tetraol, psoralen, isopsoraside, and secretinoside, and add methanol to make the concentrations of 2.71 and 8.61, respectively. , 7.61, 643, 17.42, 11.88, 45.00μg/mL mixed reference solution.

1.2.3 Preparation of the test solution

Precisely weigh an appropriate amount of each medicinal material according to the prescribed amount (15g antler, 9g safflower, 20g psoralen, 30g epimedium, 15g dogwood meat, 30g Ligustrum lucidum, 6g agarwood), add 12 times of water and cook for 75min, filter After passing, the filtrate was concentrated under reduced pressure, transferred to a 500 mL volumetric flask, adjusted to volume, shaken well, filtered through a 0.45 μm microporous membrane, and the subsequent filtrate was obtained.

1.2.4 Investigation of Linear Relationship

Take the mixed reference solution under item "1.2.2", dilute and shake well, and inject 10 μL of sample under the chromatographic conditions of item "1.2.1" for determination. Take the solution mass concentration as the abscissa (X) and the integral value of the peak area as the ordinate (Y) for regression, and draw a standard curve. The results are shown in Table 1.

Table 1 The results of the linear relationship investigation of the 7 components in the anti-chronic heart failure drug compound

1.2.5 Precision test

Take high (H), medium (M) and low (L) mass concentration reference substance solutions, inject 6 times under the chromatographic conditions of "1.2.1" within the same 1d, and measure continuously for 3 days, and measure the peak area RSD of each component. It shows that the intraday precision of the instrument is good, and the results are shown in Table 2.

Table 2 The results of the precision investigation of the 7 components in the anti-chronic heart failure drug compound

1.2.6 Repeatability test

The test solution was prepared according to the method under "1.2.3", 6 parallels were prepared, and the samples were injected and measured under the chromatographic conditions of 1.2.1, and gallic acid, hydroxysafflor yellow A, loganin, agarwood tetraol were measured. The RSDs of the contents of , psoralen, isopsoraside, and secretinoside are 2.04%, 2.78%, 2.51%, 2.78%, 0.75%, 2.49%, and 0.96%, respectively, indicating that the method has good repeatability. Test requirements.

1.2.7 Stability test

Prepare the test solution according to the method under item "1.2.3", inject samples at 0, 1, 4, 6, 12, 16, and 24 h under the chromatographic conditions of item 1.2.1, and measure gallic acid, hydroxysafflor yellow The RSDs of the peak areas of pigment A, loganin, agarwood tetraol, psoralen, isopsoraside, and teruquitoside were 1.6%, 2.7%, 2.5%, 2.9%, 0.9%, 1.8%, and 1.3, respectively. %, indicating that the solution has good stability within 24h.

1.2.8 Sample addition recovery test

Precisely measure 5mL of the test solution with known content under "1.2.3", put it in a 10mL volumetric flask, add the reference solution at 50%, 100%, 150% level, under the chromatographic conditions of 1.2.1 The sample was injected and measured, and the recovery rate was calculated. As a result, the average loading recoveries of gallic acid, hydroxysafflor yellow A, loganin, agarwood tetraol, psoralen, isopsoraside, and estrogenin were 102.58%, 98.85%, and 100.17%, respectively. , 98.87%, 98.61%, 97.32%, 98.53%, and the RSDs were 3.72%, 3.08%, 3.92%, 2.02%, 1.21%, 2.02%, 1.14%, respectively. The results are shown in Table 3.

Table 3 The recovery rate test of sample addition for the determination of the content of the index components in the anti-chronic heart failure drug compound

1.2.9 Sample determination

Prepare the test solution according to the method in Item 1.2.3, inject the sample under the chromatographic conditions of Item 1.2.1, and see the results in Table 4.

Table 4 Determination of the content of 7 index components in the anti-chronic heart failure drug compound (n=3)

2 Discussion

(1). Chronic heart failure can be divided into 4 stages A, B, C, and D according to the severity ^[15] . Among them, the traditional Chinese medicine treatment in stage B is still in the blank period of the market, and clinical application and animal tests have proved that it is resistant to chronic heart failure. Drug compounds can not only improve symptoms, improve quality of life, but also delay the development of myocardial remodeling. The prescription is composed of 7 herbs: antler, safflower, epimedium, psoralea, privet japonica, dogwood and agarwood. Activating blood and clearing the meridians, the two medicines are the king medicine; Epimedium, Psoralea, Ligustrum lucidum, and Cornus help the antlers to calm the yin and yang of the kidney, and they are the minister medicine; Agarwood absorbs the qi and returns to the kidney to help the kidney medicine, and promotes qi and relieves pain. Helping safflower to activate blood circulation, and make this prescription tonic without stagnation, it is an adjuvant medicine.

(2). According to the 2015 edition of the Chinese Pharmacopoeia, hydroxysafflorin A, psoralen, isopsoralen, icariin, loganin, morroniside, and quinoside are all corresponding medicinal materials The quality control index of agarwood tetraol; agarwood tetraol as the main medicinal component in agarwood, the 2015 edition of "Chinese Pharmacopoeia" for the first time determined the content of agarwood tetraol as a quality control index ^[16] , agarwood has a therapeutic effect against arrhythmia and coronary heart disease, The quantitative analysis of the content of agarwood tetraol as the index component is an important part of establishing the effective quality evaluation system of agarwood. According to the previous experimental results, the contents of psoralen, isopsoralen, icariin and morroniside in the decoction of the anti-chronic heart failure drug compound are relatively low, and they should not be used as the quality control indicators of the compound. Liposide and isopsoraside can be converted into psoralen and isopsoralen in vivo ^[17] , and the content in decoction is high, so gallic acid, hydroxysafflor yellow A, Loganin, agarwood tetraol, psoralen, isopsoraside, and tequilagenin were used as the quality control indicators of the anti-chronic heart failure drug compound.

(3). Selection of mobile phase: After comparison and screening, it was found that the 0.1% formic acid-acetonitrile system was eluted with a linear gradient, and the obtained liquid chromatogram had a relatively stable baseline, symmetrical chromatographic peaks, and good resolution. Therefore, 0.1 was selected. The % formic acid-acetonitrile system was used as the mobile phase, and the comparison results are shown in Figure 2.

(4) Exploration of the mobile phase elution gradient: After systematic exploration and optimization, it is found that the following linear gradient elution procedure can not only detect the chemical components in the preparation more comprehensively, but also obtain a relatively ideal separation effect for these components. : The elution procedure is determined as the mobile phase is acetonitrile-0.1% phosphoric acid water, the elution gradient is 0～10min, 5% acetonitrile; 10～15min, 5%～10% acetonitrile; 15～35min, 10%～18% acetonitrile; 35～40min, 18%～25% acetonitrile; 40～60min, 25%～35% acetonitrile, the recording time is 0～60min, the comparison result is shown in Figure 3.

(5) Selection of chromatographic wavelength: The comparison and analysis of the three-dimensional spectrum of liquid chromatography-ultraviolet spectrum shows that the baseline is uneven and the separation degree is poor at 211nm and 330nm; the baseline is relatively flat in the preparation atlas of 254nm, 265nm, 280nm and 300nm. , the peak shape is acceptable, but the number of peaks is too small, which cannot fully reflect the composition of the preparation. The baseline is relatively flat at 230 nm, except for the known components of gallic acid, hydroxysafflor yellow A, loganin, agarwood tetraol, tonic In addition to the good detection of skoraloside, isopsoraside, quincetin, quercetin, etc., the fingerprint peak resolution of other components is also good. Considering the number of chromatographic peaks, the baseline and the corresponding strong signal Weak, 230nm was determined to be the best detection wavelength for the fingerprint of the preparation. Compared with other wavelengths, this wavelength has more peaks, and the response and resolution of each component are good. The comparison results are shown in Figure 4.

references

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[2] QU H, WANG Y, WANG Y, et al.Luhong formula inhibits myocardialfibrosis in a paracrine manner by activating the gp130/JAK2/STAT3 pathway incardiomyocytes[J].J Ethnopharmacol, 2017, 202(56):28-37 .

[3] Xu Jijie, Qu Huiyan, Rong Jingfeng. Effects of Luhong Granules on hemodynamics and plasma angiotensin II and aldosterone in rats with heart failure [J]. Journal of Shanghai University of Traditional Chinese Medicine, 2015, 29(05):53- 56.

[4] Xu Jijie, Qu Huiyan, Rong Jingfeng, etc. Effects of Luhong Granules on ACE2-Ang(1-7) axis in rats with heart failure [J]. Journal of Shanghai University of Traditional Chinese Medicine, 2015, 29(6): 41-44, 49.

[5] Huang Xiaowei, Xu Yan, Han Dong. Effects of antler polypeptide-mediated differentiation of myocardial stem cells on the expression of terminal myocardial differentiation genes ANP and MLC-2v [J]. Journal of Jilin University (Medical Edition), 2018, 44(02) :249-253+464.

[6]Liu Y N.Zhou Z M.Chen P.Evidencethat hydroxysafflor yellow Aprotects the heart against ischemia-reperfusion injury by inhibitingmitoehondrial permeability transition pore opening[J].Clinical andExperimental Pharmacology and Physiology,2008,35(2):211- 216.

[7]Zhu H B.Zhang L.Wang Z H, et al.Therapeutic effects of hydroxysafflor yellow A on focal cerebral ischemic injury in rats and itsprimary mechanisms[J].Journal of Asian Natural Products Research.2005,7(4):607 -613.

[8]ZHOU H, YUAN Y, LIU Y, et al.Icariin attenuates angiotensin IIinduced hypertrophy and apoptosis in H9c2 cardiomyocytes by inhibiting reactiveoxygen species-dependent JNK and p38 pathways[J].Exp Ther Med, 2014, 7(5):1116 -1122.

[9]ZHANG Q, LI H, WANG S, et al.Icariin protects rat cardiac H9c2 cells from apoptosis by inhibiting endoplasmic reticulum stress[J].Int J Mol Sci, 2013, 14(9):17845-17860.

[10] Xiang Yang, Ma Long. Effects of grape skin and seed extracts on blood lipid levels in hyperlipidemia rat model [J]. Journal of Xinjiang Medical University, 2005(6):521-523.

[11] Hu Wanying, Zhou Hua, Dong Yaorong. Clinical and experimental research on the treatment of heart failure with Lujiao Fang [J]. Shanghai Journal of Traditional Chinese Medicine, 1992(11):8-11.

[12] Feng Xiaopeng, Zhao Wenwang, Pi Wenxia, et al. Protective effect of the active ingredients of Cornus officinalis on the myocardium of diabetic mice [J]. Chinese Journal of Gerontology, 2016, 36(19): 4681-4682.

[13] Wang Huishu, Liu Yulan. The effect of oleanolic acid on platelet function [J]. Journal of Shenyang Pharmaceutical University, 1993, 10(4):275.

[14] Zhang Anna, Li Peng, Hong Xiaohua, Chen Feiyu, Wang Yanghui, Liu Xinyu, Ren Junguo, Liu Jianxun. The protective effect of tereprivetin on the apoptosis of mesangial cells stimulated by high glucose [J]. Chinese Experimental Prescription Journal of Science, 2015, 21(08):116-119.

[15] Dai Guizhu. 2005 American Chronic Systolic Heart Failure Treatment Guidelines Analysis [J]. Chinese Journal of Cardiovascular Diseases, 2005, 33(12): 1065.

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[17] Di Qingfeng. Study on the extraction process of active components of Psoraleae [J]. Guangdong Chemical Industry, 2019, 46(23): 45-46.

The above are only the preferred embodiments of the present invention. It should be pointed out that for those skilled in the art, without departing from the method of the present invention, several improvements and supplements can be made, and these improvements and supplements should also be regarded as It is the protection scope of the present invention.

Claims (8)

1. A high performance liquid chromatography method for simultaneously determining the content of 7 components in an anti-chronic heart failure medicine compound is characterized in that the 7 components are gallic acid, hydroxysafflor yellow A, loganin, agaroterol, psoralen glycoside, isopsoralen glycoside and specnuezhenide, and the chromatographic conditions of the high performance liquid chromatography method are as follows: adopting a C18 chromatographic column, wherein the mobile phase acetonitrile-0.1% phosphoric acid water is adopted, the elution gradient is 0-10 min, and the elution gradient is 5% acetonitrile; 10-15 min,5% -10% acetonitrile; 15-35 min,10% -18% acetonitrile; 35-40 min,18% -25% acetonitrile; 40-60 min, 25-35% acetonitrile; the flow rate is 0.8-1.2 mL/min < -1 >, the column temperature is 28-32 ℃, the detection wavelength is 220-360 nm, and the anti-chronic heart failure compound traditional Chinese medicine comprises the following components in percentage by weight: 10-20 parts of antler, 6-12 parts of safflower, 15-25 parts of fructus psoraleae, 25-35 parts of epimedium herb, 10-20 parts of cornus officinalis, 25-35 parts of glossy privet fruit and 4-8 parts of agilawood, and the preparation method of the test solution by the high performance liquid chromatography method comprises the following steps: precisely weighing the above materials, adding 8-15 times of water, decocting for 60-90min, filtering, concentrating the filtrate under reduced pressure, transferring to volumetric flask, metering volume, shaking, filtering with microporous membrane, and collecting the filtrate.

2. The high performance liquid chromatography method according to claim 1, wherein the mixed reference solution is prepared by: accurately weighing gallic acid, hydroxy safflower yellow A, loganin, linalool, psoralen, isopsoralen, and specnuezhenide, and adding methanol to obtain mixed reference solutions with concentrations of 2.71, 8.61, 7.61, 643, 17.42, 11.88, and 45.00 μ g/mL.

3. The high performance liquid chromatography method according to claim 1, wherein the amount of sample is 1 to 12. mu.L.

4. The high performance liquid chromatography method according to claim 3, wherein the amount of the sample is 10. mu.L.

5. The high performance liquid chromatography method according to claim 1, wherein the flow rate is 1.0mL _ min "1.

6. The high performance liquid chromatography method of claim 1, wherein the column temperature is 30 ℃.

7. The high performance liquid chromatography method according to claim 1, wherein the detection wavelength is 220 to 240 nm.

8. A quality control method of a compound anti-chronic heart failure medicine, which is characterized by comprising the steps of the high performance liquid chromatography method of any one of claims 1 to 7.

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