Internship Report by Moyan Jia

Offer: Inner Mongolia DATANG Medicine Company Ltd.

Address: Ruyihe Street, Hohhot, Inner Mongolia, China
Tel: +86 0471-4611148

Zip Code: 010010

Fax: +86 0471-4611132

Website: www.nmdt.com.cn
Department: Product Technology Center and Microbiology Laboratory
Position: Researcher and Quality Tester
Date: 07/18/2016-08/12/2016
Job Description: Hippophae Syrup research and quality test
The first week, getting familiar with company and managers. Getting into workshop
and supervising equipment operations. The second week to last week, participating
product registration department and technology center for new medicine research
hippophae syrup.

1.0 Introduction
1.1 Background
In traditional Chinese pharmacology, hippophae (Hippophae rhamnoides Linn) is
treated as both medical material and health care product. Hippophae juice in the
market are targeted for patients with cough and bronchitis [1]. In order to prove the
curative effects of hippophae preparations, DATANG makes efforts on the research of
recipe and technology of hippophae syrup.
Fructus Hippophae contains vitamin C and seabuckthorn flavone. Flavone
demonstrates physical characteristic like fine yellow powder; it tastes bitter, easily
dissolves in polar solvents like water and ethanol, and barely dissolves in organic
solvents [2][3]. The main ingredients of seabuckthorn flavone are isorhamnetin,
isorhamnetin-3-O--D, isorhamnetin-3-B-rutinoside, and oligosides [3].

Figure 1: isorhamnetin

Figure 2: isorhamnetin-3-O--D

Figure 3: oligosides (Kaempferol-3-O-glucoside)

They have clinic effects on cough relief, blood circulation activation, blood lipid
metabolism, and vessel softening [3].
1.2 Effective Material Contents
The hippophae juice in the market has concentration >30%, and the flavone is not
detected. According to Chinese Pharmacopeia, medical hippophae syrup must contain
rutinum (C27H30O16) 1.5%, and isorhamnetin (C16H12O7) 0.10%, which are the
major components of flavonoids [3].
2.0 Materials and Methods
2.1 Hippophae Juice Preparation and Measurements
At the very initial stage of new medicine research, experimenters directly squeeze the
fructus hippophae to make syrup. Experimenters used the drying-weight method to
measure concentration (130, 1 hour). However, the concentration is as low as 17%.
2.2 Hippophae Extraction
Then experimenters used orthogonal experimental design to make the ethanol
extraction. Experimenters mixed squeezed juice with ethanol in the beaker. The
mixture was shaken well and filtered. Then 100% ethanol was added into the filter
liquor, centrifuged multiple times (5 min, 2800rpm) , and the solution contained in
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flask was put into 4 fridge for period . Then experimenters proceeded suction
filtration and put the liquid into drying cabinet at certain temperature .
2.3 Orthogonal Experiment for Extraction
Table 1: Orthogonal experimental design: Lg(34)
Elements
Ethanol

Period

Temperature

60%

8 hours

50

70%

10 hours

60

80%

12 hours

70

Levels

Table 1: Orthogonal experiment

Elements
Ethanol

Period

Temperature

60%

8 hours

50

60%

10hours

60

60%

12 hours

70

70%

8 hours

60

70%

10 hours

70

70%

12 hours

50

80%

8 hours

70

80%

10 hours

50

80%

12 hours

60

Rutinum
Concentration/mg/50ml

Numbers

To be measured

2.4 Extractive Contents Measurements

Seabuckthorn flavonoids dissolved in ethanol react with Al3+ in weak base, and the
product can be detected by UV spectrophotometer. It has the highest absorbance at
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wavelength 415 nm. According to the previous study, the absorbance is positive
related to falvonoids concentration.
3.0 Results and Discussion
The 9 flasks labeled as the orthogonal experiment above were measured flavonoids
contents. Since the experiment was at the early stage and still in process, the data was
not collected yet.
Reference
Figure 1: http:// www.chembase.fudan.edu.cn%2Fsubstance-361493.html
Figure 2: http://www.chem17.com
Figure 3: http://www.instrument.com.cn/netshow/SH103151/Q4161166.htm
[1] Renzhen, Wang-jia., Literature Research of Tibetan medicine Hippophae
rhamnoides, Chinese Folk Herbs, 2015, 25(6)
[2] Li, li-jun., Chen, Jiang-mei., He, Jian-chao.,Evaluation of uncertainty in
determining the total flavonoids of sea buckthorn juice, Journal of Northwest
University (Natural Science Edition), 2010, 40(6)
[3] Sang., Guo-wei, Hippaphae Fructus, Pharmacopeia of People s Republic of China,
2015 Edition, Volume 1, pg 184-185.

Experiment steps
1. Preparation
Wash new glass containers:
Soak in acetic acid for 4 hours
Wash by 2% HCl or washing liquid
Wash by water
Wash used glass container:
Sterilize 121 30min
Pour out the media
Wash by water and detergent
Wash by purified water
Airdry on the gridding shelf
Wash old pipes:
Sterilize in 3%-5% Lysol or 0.25% bromoeramine or 5% phenol contained in
glass jar for 24 hours, 0.1MPa sterilize for 30 min.
Wash by flowing water, wash the cottons off
Soak in detergent waster for a night
Wash by purified water
Dry in the dryer
Clean Petri dishes are put into the breathing bag, and sterilize
Put the cotton into pipes
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Put the silicon rubber tips into conical flasks and seal by tinfoil
2. Sterilization
Glass containers 160 2 hours
Tweeters 121 30min
Rubber tips 75% ethanol soaked
3. Experiments
Put the cleanroom clothes, respiration mask rubber gloves.
Deliver samples and experiment equipment from delivery window
Sterilize the clean bench
Weigh the samples and put them into media and oscillator
Pour certain amount sample and media into Petri dishes
(Mycete Rose Bengal Agar Medium)
(Bacteria Trypose Soya Agar/Liquid Medium)
Put the prepared Petri dished into incubator
Se temperature, time, and humidity
Count cfu.(Colony Forming Units)