Study on Preparation Process of Shanxian Zaopi Granules

Han Zhou, Xia Dong, Bobo Meng, Xiaoping Wang and Yan Fang

Shaanxi University of Chinese Medicine, Xianyang, 712046, China

Keywords:

Shanxian Zaopi Granules, High-Performance Liquid Chromatography, Excipients, Dry Paste.

Abstract:

Objective: To establish the preparation method and quality standard of Shanxian Zaopi granules. Methods:

High-performance liquid chromatography was used to optimize parameters including granule appearance,

formation ability, dilution rate, solubility and content of forsythia glycosides, and the type and ratio of

auxiliary materials. Results: The excipients were synthesized into granules using dextrin and starch at the ratio

of 2:3. The optimum ratio of the dry paste and excipients was 1:1.5. Curcuma oil and an equal amount of

anhydrous ethanol were evenly sprayed onto the granules. Conclusion: The methods for the preparation and

quality control of Shanxian Zaopi granules are simple, feasible and reliable. The granules have good properties

and conform to the relevant provisions of the Pharmacopoeia of China.

1 INTRODUCTION

Breast cancer is the most prevalent malignancy in

women and the second leading cause of cancer-

related deaths worldwide (Tong, 2021). Around 2.3

million new cases of breast cancer were reported

globally in 2020, with a mortality rate of 6.9% (Sung,

2021). The current treatment modalities including

surgery, radiotherapy, chemotherapy, targeted

therapy etc. (Cao, 2021) have failed to improve the

long-term survival rate substantially (Shen, 2008),

and are associated with an increased risk of

secondary malignancy (Tong, 2021). In addition, the

incidence of postoperative complications is also high

at 33% (Nicotera, 2021). The adoption of several

unhealthy lifestyle habits in recent years have

increased the risk of postmenopausal breast cancer by

more than three-fold and the risk of death by 96%

(Lofterød, 2020). precancerous breast cancer are

breast lesions that may develop into cancer but are

not cancerous at present, such as breast enlargement,

breast nodules, breast cysts, mastitis, and so on. If left

untreated for a long time, they may develop into

tumors. Pre-cancerous breast lesions found should be

treated actively to prevent development to breast

cancer. Only a minority of women progress from

hyperplasia to atypical hyperplasia and carcinoma in

situ. If a patient develops atypical hyperplasia, there

is a risk of developing cancer. The treatment of pre-

cancerous breast cancer in Western medicine is still

mainly based on surgery, and how to effectively

interrupt the development of pre-cancerous disease

has not been given enough attention (Deng, 2013).

Therefore, the prevention of breast cancer becomes a

major clinical challenge to be overcome. Early

diagnosis and treatment of breast cancer will greatly

improve the survival rate of patients.

Professor Cai Guoliang from our university and

Professor Ma Shuanquan, a doctor of Chinese

medicine at our affiliated hospital in Shaanxi

Province, have developed the "Zaoqiao Xiaopi”

prescription based on the "Zhitong Xiaojie”

decoction for treating precancerous breast cancer.

The formulation consists forsythia, Spina Gleditsiae,

Rhizoma Zedoariae, Fritillaria thunbergii, Rhizoma

Corydalis and bupleurum. Spina Gleditsia can

penetrate pus, break firmness, eliminate

accumulation and relieve pain, whereas forsythia can

clear heat and detoxify, dispel blood stasis and reduce

swelling in skin infections. The combination of Spina

Gleditsiae and forsythia can clear away heat,

detoxify, eliminate accumulation, relieve pain, and

alleviate hypochondriasis (Meng, 2018). In addition,

both herbs have documented anti-tumor, anti-

inflammatory, antibacterial, antioxidant and other

pharmacological effects (Zhai, 2021; Xing, 2017).

Rhizoma Zedoariae can soothe the liver and promote

Qi flow, break blood and expel stasis. Curcumol, a

bioactive component of the volatile oil of Rhizoma

Zedoariae, promotes tumor cell apoptosis and inhibits

Zhou, H., Dong, X., Meng, B., Wang, X. and Fang, Y.

Study on Preparation Process of Shanxian Zaopi Granules.

DOI: 10.5220/0012013600003633

In Proceedings of the 4th International Conference on Biotechnology and Biomedicine (ICBB 2022), pages 82-85

ISBN: 978-989-758-637-8

 2023 by SCITEPRESS – Science and Technology Publications, Lda. Under CC license (CC BY-NC-ND 4.0)

proliferation and invasion (Nie, 2020). F. thunbergii

can dissipate phlegm and blood stasis, and augment

the action of Rhizoma Zedoariae. Verticine and

verticinone, the effective active components of F.

thunbergii, can reverse chemoresistance of tumor

cells and act synergistically with other anti-tumor

drugs (Zhao, 2019). Rhizoma Corydalis is used as an

adjuvant to promote the flow of Qi, activate blood

circulation, and relieve pain. Bupleurum is used to

soothe the liver and circulate Qi. According to some

researches, Rhizoma Corydalis also has anti-

arrhythmic, anti-bacterial, anti-inflammatory and

anti-tumor effects, and has protective effects on

cardiovascular system, respiratory system, central

nervous system, and so on (Wang, 2010). Therefore,

the combination of these herbs can remove heat

toxins, eliminate blood stasis and phlegm, relieve

sclerosis, and target the pathogenesis and cure the

root cause (Li, 2016). In this study, the herb

formulation was synthesized into a granular form in

order to simplify administration. We optimized the

preparation process and the number of auxiliary

materials, and established a method for quality

control.

2 INSTRUMENTS AND

MATERIALS

2.1 Instruments

Agilent 1260 high-performance liquid

chromatography (HPLC) system, Agilent Zorbax

C18 column (4.6 mm × 150 mm, 5 μm), GHX-

9270B-1 Thermotank (Fuma, Shanghai, China),

DHX-9140A Drying Cabinet (Jinghong, Shanghai,

China), HEALFORCENW10UF Water Purifier

(Hong Kong), HH-S Digital-displayed Thermostat

Water Bath Cauldron (Jiangsu Jintan Medical

Instrument Factory), JFSD-100 Crusher (Shanghai

Jiading cereals and oils Instrument Co., Ltd.), and

Medicine Sifter 3, 4 and 5 were used in this study.

2.2 Materials

Rhizoma Zedoariae (10 g) was suspended in 300 mL

water in a flask, and several glass beads were added.

The mixture was heated on an electric heating mantle

connected to volatile oil detector and reflux

condenser. The solution was brought to boil and then

simmered for about 5 hours until the amount of oil in

the detector no longer increased. The heater was

switched off, and the plant matter was stewed. The

water was slowly released till the oil rose to the

topmost layer and flushed with Scale 0. The amount

of volatile oil was calculated to be approximately 2%.

Nine batches of the herbs were weighed according to

the prescription (15 g forsythia, 12 g Spina Gleditsia,

15 g Rhizoma Zedoariae, 15 g F. thunbergii, 12 g

Rhizoma Corydalis and 12 g bupleurum). The herbs

were soaked in 6 volumes of water for 30 min, and

decocted and extracted twice for 30 min each time.

Approximately 100 mL of the extract was collected,

and an equal amount of 95% ethanol was added. The

solution was allowed to rest for 24h, and the sediment

was boiled, concentrated and reconstituted to 100

mL. The samples (10µl aliquots) were analyzed by

HPLC using Agilent Zorbax C18 column (4.6 mm ×

150 mm, 5μm) with acetonitrile-water (25:75) as the

mobile phase. The detection wavelength was 277 nm,

the detection temperature was 30℃, and the flow rate

was 0.1 mL/min. The content of forsythia glycosides

in the solution was determined to be 0.0516 mg/mL.

The solution was heated and concentrated, and the

dry paste was obtained. The paste rate was calculated

as dry paste amount (g)/amount of herbal matter (g).

Sucrose (batch number: 10012020200802) was

provided by Nanning Guizhilong Pharmaceutical raw

materials Co., Ltd. Dextrin (batch number: 140310)

and soluble starch (batch number: 131209) was

obtained from Shaanxi Mr G Pharmaceutical Raw

Materials Co., Ltd.

3 METHODS AND RESULTS

3.1 Preparation Method

3.1.1 Selection of Excipients

Sucrose, dextrin and soluble starch are commonly

used as excipients for granules (Huang, 2016).

However, given the viscous nature of sucrose, it is not

suitable for the granulation of water extract

containing polysaccharides. Therefore, we used a

mixture of dextrin and soluble starch to form the

granules, and the appropriate proportion of the

excipients was selected according to the formation

ability and moisture absorption rate.

3.1.2 Amount of Excipients

The drug solution was extracted as previously

described. Briefly, the herbs are soaked in 6 times the

amount of water for 30 min, decocted and extracted

Study on Preparation Process of Shanxian Zaopi Granules

twice for 30 min each time, and dried into a paste.

Different amounts of soluble starch and dextrin were

added and mixed evenly to make the granules

(Zheng, 2016).

3.2 Results

3.2.1 Ratio of Excipients

The granules were prepared by mixing dextrin and

soluble starch at the ratio of 2:3, and showed a high

formation rate and low moisture absorption rate

(Table 1).

Table 1: Ratio of excipients ratio.

Excipient Granule formation

Moisture absorption

rat

Dextrin Fragile granules 11.2

Starch Poor formation and fragile granules 12.4

Dextrin: Starch = 3:2

Different sizes of granules can be synthesized and

easil

broken into

owder for

10.5

Dextrin: Starch = 2:3 Good color and formation 7.8

Dextrin: Starch = 1:1

Size of the granules is not uniform

8.1

3.2.2 The Ratio of Dry Paste and Excipients

Granulation and the characteristics and solubility of

the granules were used as indicators to compare the

amount of excipient. The optimum ratio of dry paste

and excipient was 1:1.5 (Table 2).

Table 2: Ratio of dry paste and excipients.

Ratio of dry paste and excipients Granulation Solubility

1:1

Sticky, not easy to pass through

the sieve

Good solubility

1:1.3 Easier to pass through the sieve Good solubility

1:1.5 Easy to pass through the sieve Good solubility

3.3 Process Validation

The volatile oil of Rhizoma Zedoariae (19 ml from

1,500 g herb) was extracted and mixed with 6,600 g

forsythia, Spina Gleditsia, F. thunbergii, Rhizoma

Corydalis and bupleurum. The herb mixture was

soaked in 6 volumes of water for 30 min, heated, and

extracted twice for 30 min each time. Around 5,690

mL of the extract was mixed with an equal amount of

95% ethanol and left undisturbed for 24 h. The

supernatant was discarded and the sediment was

boiled and concentrated into a paste. About 720 g

(612 mL) dry paste was obtained, which was then

crushed, passed through sieve 5, and mixed with 430

g dextrin and 650 g starch to make the granules. In

addition, 19 mL each of Rhizoma Zedoariae volatile

oil and anhydrous ethanol were evenly sprayed onto

the granules. After drying in an oven at 40℃ (24 h),

1,800 g dried granules were obtained and packed into

lots of 9 g. Three such batches were prepared, and 5

samples were randomly selected from each batch.

The contents of each were dissolved in warm water

to a final volume of 50 ml, and 10μL aliquots were

taken for HPLC. The average content of forsythia

glycosides was 0.0513 mg/mL. The results were

stable.

4 CONCLUSION

Shanxian Zaopi granules are composed of six herbs,

namely forsythia, Spina Gleditsiae, Rhizoma

Zedoariae, Fritillaria thunbergii, Rhizoma Corydalis

and bupleurum. The combination of these herbs can

remove heat toxins, eliminate blood stasis and

phlegm, relieve sclerosis, and target the pathogenesis

and cure the root cause. Therefore, the granules can

effectively treat precancerous breast cancer for

clinical treatment. However, there are fewer studies

on other aspects of the effects. The bioavailability is

ICBB 2022 - International Conference on Biotechnology and Biomedicine

low, and the side effects of the granules are still

unclear, so how to apply it more safely and efficiently

in clinical treatment is an urgent problem for

researchers to solve at present.

In this study, we used high performance liquid

chromatography (HPLC) to determine the content of

forsythia glycosides, the main active ingredient of

forsythia, which can better control the quality of

Shanxian Zaopi granules. HPLC has many

advantages, such as high separation efficiency, high

detection sensitivity, good selectivity, automated

operation, and wide range of applications. However,

it also has the disadvantage of high analysis cost,

expensive liquid chromatograph price and routine

maintenance. With the rapid development of Chinese

medicine preparation production technology, aim to

achieve easier and more sensitive quality control of

formulated particles, Some new analytical techniques

such as infrared spectroscopy, gas chromatography,

high performance liquid chromatography and other

qualitative and quantitative detection methods have

emerged. Quality control of formulated particles

gradually becomes the focus of researchers'

attention.The preparation process of granules is

complicated. How to improve the preparation

technology and perfect the preparation process is a

problem that researchers need to consider nowadays.

Meanwhile, the extraction process of the granules

also needs to be paid attention to, on the basis of

which the preparation process will progress

smoothly.

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Study on Preparation Process of Shanxian Zaopi Granules