This document discusses phytopharmaceuticals and carotenoids. It defines phytopharmaceuticals as purified plant-derived medicinal drugs. Carotenoids are plant pigments that give fruits and vegetables their red, orange, and yellow colors. They are unsaturated hydrocarbons found in photosynthetic bacteria, fungi, algae, plants, and some animals. The document describes the isolation of carotenoids from carrots using extraction and column chromatography. It also outlines the characteristic features and medicinal uses of carotenoids, which include their antioxidant and anticancer properties.

1. Phytopharmaceuticals- Carotenoids
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MAULANA AZAD UNIVERSITY, JODHPUR

2. Contents
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Introduction of phytopharmaceuticals
Carotenoids
• Occurrence
• Isolation
• Characteristic features
• Medicinal uses
References

3. Introduction
• Phyto- referring to plant, pharmaceutical- refers to
medicinal drugs
• Phytopharmaceutical drug is defined as purified and
standardized fraction with defined minimum four
bio-active or phytochemical compounds of an
extract of a medicinal plant or its part, for internal
or external use of human beings or animals for
diagnosis, treatment, mitigation, or prevention of
any disease or disorder but does not include
administration by parenteral route.[1]
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4. Disadvantages:
 USFDA estimated over
50,000 adverse events
caused by botanical and
other.dietary supplements.[1]
 Use of incorrect plant
species
4
Advantages:
Less side effects as compared to synthetic
drugs
Trust
Availability
Standardization
Insolubility, hydrophobicity,
low bioavailability, and high
toxicity (in some plants)
Lack of knowledge among
people.

5. Licensing of a patent or proprietary
phytopharmaceuticals
5
In 2010
Rule 158 (B)
By AYUSH
Proof of
effectiveness
Patent or
proprietary ASU
medicine[1]
Patent or proprietary
medicine
Ingredients
Formulations of
authoritative texts
Intellectual
intervention,
innovation, or
invention
Unique medicine[1]
Issue of license

6. • Data requirements specified in Appendix IB of
Schedule Y of D&C Rules.
• Clinical trials conducted as per applicable rules and
guidelines for new drugs.
• For all phytopharmaceutical drugs data from phase I
(to determine maximum tolerated dose and associated
toxicities) and the protocols shall be submitted prior
to performing the studies.
• Data of results of dose finding studies performed and
the protocols shall be submitted prior to performing
the studies.[3]
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Does a new Phytopharmaceutical drug
undergo clinical trial in India?

7. CAROTENOIDS
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8.  Characteristic features
• They are tetraterpenoids consisting of
isoprene (C5H8)n units.
• Carotenoids are plant pigments with ,
, and colors.[5]
• These are chemically unsaturated
hydrocarbons.[6]
• They are strongly lipophilic in nature.
• Long series of conjugated double bonds
forming the central part of the molecule.
• This gives them their shape, chemical
reactivity, and light-absorbing properties.[5]
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9. Classification of carotenoids
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10. Chemical structures of (a) α-carotene, (b) β-carotene
and (c) lutein.
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11.  Occurrence
• About 850 naturally occurring carotenoids had
been reported up until 2018.[2]
• Found in:-
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Human beings
Can’t synthesize
Photosynthetic bacteria,
fungi, algae, plants and
some animals.
Synthesize carotenoids
De Novo

13.  Isolation[4]
13
Extraction
of carrots
Cold
maceration
n-Hexane +
Acetone
(1:1)
+BHT and
BHA
(antioxidants)
505 mg crude
carotenoid
extract obtained
from 1 kg
CCC 1000 instrument by Pharma-Tech Research can be used for the separations.
Solvent
system
Two
phase
n- Hexane :
Benzotrifluoride :
MeCN (10 : 3.5 : 6.5)
Transfer to
separating
funnel
Shake
vigorously
Stand for 24 hours, then separate upper and lower phase and degass both for 10 min
in ultrasonic bath.

14. 14
Sample
solution
100.2 mg crude extract + 10 ml of mixture of upper and lower
phase (1:1)
Dual mode CCC separation was performed at ambient lab temperature (22ºC)
Tail to head
mode
Fill Separation
column with
lower phase
Adjust flow
rate
Start
centrifuge
rotation
Pump upper
phase into the
apparatus
Collect
effluent
Mobile phase
emerged at
outlet
Measure displaced
volume of
stationary phase
Carry out
sample
injection
Separation
conducted for
140 mins.
Complete elution of α-carotene
according to UV-vis
spectrophotometer

15. Reverse
column inlet
& outlet
Select head to
tail mode
Pump lower
phase into the
head for 60
min
Elution of
Lutein
β-Carotene
collect
fractions from
95-105 min
1fraction,
20ml
α-Carotene
collect fractions
from 115- 130
min
At 2.5
min/tube
6 fractions,
5ml each
Lutein
collect fractions
from 160- 172
min
At
2.0min/tu
be
6 fractions,
8ml each

16.  Medicinal uses[8]
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17. 17

18. References
1. Bhatt A. Phytopharmaceuticals: A new drug class
regulated in India. Perspect Clin Res. 2016 Apr-
Jun;7(2):59-61. doi: 10.4103/2229-3485.179435.
PMID: 27141470; PMCID: PMC4840792.
2. Maoka T. Carotenoids as natural functional
pigments. J Nat Med. 2020 Jan;74(1):1-16. doi:
10.1007/s11418-019-01364-x. Epub 2019 Oct 1.
PMID: 31588965; PMCID: PMC6949322.
3. https://www.cdsco.gov.in/opencms/export/sites/CDS
CO_WEB/Pdf-documents/New-
Drugs/FAQs/New_Drugs_FAQs.doc
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19. 4. Englert M., Hammann S., Vetter W. Isolation of β-
carotene, α-carotene and lutein from carrots by
countercurrent chromatography with the solvent system
modifier benzotrifluoride, J. Chromatogr. A, 1388, 2015,
119-125, ISSN 0021-9673.
5. Miller D.D., Li T., Liu R.H., Antioxidants and
Phytochemicals, Reference Module in Biomedical
Sciences, Elsevier, 2014, ISBN 9780128012383.
6. Marzena B., Paulina M., Katarzyna B. Characteristics of
carotenoids and their use in the cosmetics industry. J.
Educ. Health Sport. 2020. 10. 192-. DOI:
10.12775/JEHS.2020.10.07.020.
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20. 7. Johnson E.J. The role of carotenoids in human health.
Nutr Clin Care. 2002; 5(2):56-65.
8. Nabi F, Arain M.A., Rajput N, et al. Health benefits
of carotenoids and potential application in poultry
industry: A review. J Anim Physiol Anim
Nutr. 2020; 104: 1809– 1818.
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21. Any Questions