Experiment No.

1
PREPARATION OF CRUDE DRUG

1. What is a crude drug?

Crude medications are plant or animal substances that have simply been collected and dried;
Crude drugs are entirely natural raw substances used in medicine.

2. Where are underground parts used as drugs collected?

Underground plant parts should be collected after the vegetation period has ended to optimize
its active constituent as drug collected. The plant sample should be cleaned of any clinging soil
as soon as it is obtained to avoid contamination.

3. What is the ideal weather for collecting flowers?

Flowers are ideally collected in the morning (after the dew has evaporated) or evening and
when it is fully developed. The best time to gather flowers is early in the day when it is cool
outside, the weather is not too hot, and the plant's water level is high.

4. How are leaves used as drugs packaged and stored?

Leaves used as drugs are packed and stored in low humidity (below 10% moisture),
well-ventilated, spacious storage area. Dry drugs are stored in a tightly-sealed container and
protected from daylight. The packaging must provide ample protection from insect attacks on
the drug and give the economy of space. They may make use of air-conditioning and
humidity-control equipment.

5. Differentiate gums from latex.

Gums and latex are both sap, for they are water-soluble polysaccharides. Gum is a sap
associated with woody plants particularly in a seed coating or under the bark while the latex is a
milky fluid found in flowering plants such as the rubber tree.

6. How and when are plant exudates (gums, latex resins) collected?
Plant exudates such as gums, latex and resins are best collected during dry weather.
Plant exudates are collected from trees through tapping, which is the process of causing minor
damages or making incisions on the tree to induce release of exudate.
References: (APA 7)

Harvesting. (n.d.). KAU Agri-Infotech Portal:CEL.

https://www.celkau.in/crops/Plantation%20Crops/Rubber/harvesting.aspx
Tsmith. (2016, August 4). Harvesting and Handling Cut Flowers. Center for Agriculture, Food,
and the Environment.
https://ag.umass.edu/greenhouse-floriculture/fact-sheets/harvesting-handling-cut-fl
owers#:~:text=Flowers%20should%20be%20harvested%20in,to%20obtain%20long%2C
%20sturdy%20stems

Dowse, M. (n.d.). Sources and types of carbohydrates and sugar. Sugar Nutrition Resource
Centre. Retrieved March 7, 2023, from
https://www.sugarnutritionresource.org/the-basics/sources-and-types-of-carbohydrates-and-sug
ar

Nutrition FN 225. Types of Carbohydrates. (n.d.). Retrieved March 7, 2023, from

https://media.lanecc.edu/users/powellt/FN225OER/Carbohydrates/FN225Carbohydrates2.html

Murthy H.N. (2021, July 21). Chemical Constituents and Applications of Gums, Resins, and Latexes
of Plant Origin. Springer Link.
https://link.springer.com/referenceworkentry/10.1007/978-3-030-91378-6_1

Edomwonyi-Otu, L.C. (16, May 2020) Natural Rubber Latex and Gum Arabic: A Comparison of
Physico-chemical Properties. Preprint org. Natural Rubber Latex and Gum Arabic: A Comparison
of Physico-chemical Properties
 Experiment No. 2
MOISTURE DETERMINATION

1. Define constant weight.

From the principle of “igniting or drying to constant weight,” constant weight means that the two
consecutive weighings do not/should not differ by more than 0.50 mg per g of substance taken,
the second of which is taken after an additional period appropriate to the nature and amount
of residue for about an hour.

2. Differentiate moisture content from water content.

The key difference between moisture content and water content is that moisture content
determines the amount of water vapour and other volatile components present in a sample,
whereas water content determines the amount of water in a sample.Water content is usually
measured through titration, especially the famous Karl Fischer method.h Moisture content, by
contrast, is measured primarily through loss on drying, which essentially means the sample is
weighed, then dried or heated, and then weighed again to measure the difference.

3. What is the importance of determining the moisture content of drugs?

Determining moisture content of drugs helps with quality control. This provides data whether
there is an excessive or deficient moisture content in the drug. The amount of moisture can
influence a pharmaceutical product's physical properties, which may lead to changes in the
drug's chemical reactivity and binding properties. Knowledge about the moisture content of the
drug can be used to prevent these kinds of alterations which may improve a drug's shelf-life.

4. What is the official monograph limit for the moisture content of drugs?

Drugs Moisture content W/W⁶

Aloes Not more than 10

Starch Not more than 5

 Digitalis Not more than 15

5. Enumerate and describe/define the other USP/NF official methods of moisture

determination.
a. Method I (Titrimetric)
● Direct titration
In this method, the test sample is directly titrated with the reagent.
● Residual titration
Unlike the direct titration, it uses an excess reagent to react with the test sample,
then the unconsumed reagent is titrated with a standard solution of water in a
solvent.
● Coulometric titration
Unlike the volumetric titration, in this method, the titrant is generated
electrochemically and does not require a standard solution.
b. Method II (Azeotropic-toluene distillation)
Azeotropic distillation is a technique which uses the ability of selected
organic compounds to form binary azeotropes with water to facilitate the
separation of the compounds from a complex matrix.(<921>Water
Determination, USP)

References: (APA 7)

Constant weight. (n.d.). Retrieved February 7, 2023, from

https://www.uspnf.com/sites/default/files/usp_pdf/EN/USPNF/usp-nf-notices/usp38_nf33_

gn.pdf

Noria Corporation. (n.d.). Karl Fischer Coulometric Titration Explained and Illustrated.

https://www.machinerylubrication.com/Read/594/karl-fischer-coulometric-titration#:~:te

xt=The%20main%20difference%20between%20the,lower
SudheerKandibanda. (2016, January 2). EVALUATION OF CRUDE DRUGS.

https://www.slideshare.net/SudheerKandibanda/evaluation-of-crude-drugs

Usp31nf26s1_c921, general chapters: <921> WATER determination. (n.d.). Pharmacopeia Online,

Drug, Pharmaceutical Analysis Reference, US Pharamcopeia, British Pharmacopeia,

European Pharmacopeia.

https://www.uspbpep.com/usp31/v31261/usp31nf26s1_c921.asp

Madhu. (2020, April 24). Difference between moisture content and water content. Compare the

Difference Between Similar Terms. Retrieved February 12, 2023, from

https://www.differencebetween.com/difference-between-moisture-content-and-water-

content/
 Experiment No. 3
ASH DETERMINATION

1. What is the importance of determining the total ash and acid-insoluble ash content of
drugs? Give their possible composition.

Total ash and acid-insoluble ash is important to determine the quality and purity of a
crude drug. This is because ash is the inorganic residue that remains after water and
organic matter has been removed. This is because the organic matter is removed by
heating in the presence of oxidizing agents while the inorganic minerals are not
destroyed by heating due to their low volatility. The organic matter is removed because
these may interfere in an analytical determination of the total amount of minerals.
-Total ash content is composed of inorganic residue after water and organic matter
have been removed. This organic residue may contain extraneous solids, residues of
organometallic compounds and essential elements which include macronutrients such
as nitrogen, potassium, and calcium.
-Acid insoluble ash consists primarily of silica and silicates. It is the proportion of a sample
that is not hydrolysed by 72% sulfuric acid and is not subsequently volatilized upon the
incineration of the acid insoluble residue.

2. What temperature range does very dull redness indicate?

A very dull red heat implies a 550-700 °C temperature. It is significant to note that the
ashing temperature for biomass fuel is 500 °C, which is lower than the average ashing
temperature for coals, which is 780 °C. The explanation is that if the biomass ash is
burned at high enough temperatures, it will likely lose some of the volatile components
present in laboratory-produced ash. As a result, an ash sample that is both
unrepresentative in terms of quality and quantity is produced at high temperatures.
Furthermore, it should be apparent that fuel ash produced in a laboratory will have a
different composition from ash gathered from real-world applications.
 3. Enumerate the approximate temperature equivalents when an electronic furnace is
used in ignition.

4. What is the purpose of taring the crucible before using it in the incineration process?

Taring the crucible before using it in the incineration process will help us to obtain the
constant weight of a sample for better results and outcome. Also it will prevent errors in
getting the ash determination by means of heating the crucible to a dull redness, cooled
in the desiccator and weighed it.

5. What is the official monograph limit for acid-insoluble ash content of drugs?

The limit of acid-insoluble ash content varies between different drug products. The
difference would be influenced by the different formulations each drug contains since
there are differences in the amount of inorganic compounds each component contains.

References: (APA 7)

Ash Content Determination. (2023). Scribd.

https://www.scribd.com/document/366707475/4-Ash-Content-Determinatio

n#

Analysis Packages for Ash (Acid Insoluble). (n.d.). Celignis Analytical

https://www.celignis.com/analyte.php?value=17

Lacey, J.A., Aston, J.E. & Thompson, V.S. (2018). Wear Properties of Ash Minerals in

Biomass. Sec. Bioenergy and Biofuels, 6.

https://doi.org/10.3389/fenrg.2018.00119

Analysis of Ash and Minerals. (n.d.). Umass.edu.

https://people.umass.edu/~mcclemen/581Ash&Minerals.html
 International Organization for Standardization. (2011). Pulps - Determination of

acid-insoluble ash. https://www.iso.org/obp/ui/#iso:std:iso:776:ed-3:v1:en

Experiment No. 4
DRUG EXTRACTION: PERCOLATION METHOD

Illustration:
Draw and label the percolation set-up.

Data and Results:

Computation:

Questions:
1. What is the principle involved in the percolation method of extraction?

Percolation method is a drug extraction procedure used in extracting the soluble

constituents of a powdered drug by which the saturated solvent is constantly being
replaced by fresh solvent until the drug is completely extracted. It is dependent on the
flow of solvent through the powdered drug; because of this, the product yields greater
concentration than the other extraction process, like the maceration.

2. Define:
a. Menstruum
It is the solvent used for the extraction of crude drugs that depends on the type of
plant, nature of the bioactive compounds, and the part of the plant used.
b. Marc
Marc is the insoluble residue that remains after the extraction of a solution with a
solvent.
c. percolate
 It is to extract the soluble parts of a drug by passing a solvent liquid through it.
3. Enumerate and define the other methods of drug extraction. Give the principle involved in
each method.

4. What plant constituents are extracted by the following solvents:

a. 80% ethanol- It extracts essential oils, resins, alkaloids, glycosides, organic acids,
chlorophyll, acrid and bitter constituents, and castor oil, but not minerals, gums, or
mucilage.

b. Cold water- It extracts sugars, proteins, gums, mucilage, pectin, tannins, acids,
coloring matter, mineral salts, glycosides, some alkaloids, most alkaloidal salts, and some
essential oil very well, but is not very shelf stable.

c. Petroleum ether- It extracts showed the presence of flavonoids, saponins, and

tannins.

d. Chloroform- Palmitic acid, stearic acid, linoleic acid, and linolenic acid were the
main fatty acids found in chloroform extracts.

e. Absolute alcohol- Alcohol is a great solvent for plants since it may extract
compounds and active constituents like resins, alkaloids, and essential oils that aren't
water-soluble. Antioxidant components from many plants and plant-based foods can
also be extracted using alcohol.

f. Sulfuric ether- is useful in the extraction of substances such as alkaloids,

terpenoids, coumarins, and fatty acids.

g. Boiling water- when water solvent is subjected to heat, it can extract starches,
gums, mucilages, alkaloids, and essential oils.

h. 5% NaCl solution- It can extract coloring matters such as dyes. It can also
extract polyphenolic compounds.

i. Heating with dilute acids- Diluted acid can extract Polyphenols such as
flavonoids, phenolic acids, tannins, lignans, and coumarins.

References: (APA 7)
Abubakar, A. R. & Haque, M. (2020). Preparation of Medicinal Plants: Basic Extraction and
Fractionation Procedures for Experimental Purposes. J Pharm Bioallied Science 12(1),
1-10. https://doi.org/10.4103%2Fjpbs.JPBS_175_19

Loneman, D. M., Peddicord, L., Al-Rashid, A., Nikolau, B. J., Lauter, N., & Yandeau-Nelson,
M. D. (2017). A robust and efficient method for the extraction of plant extracellular
surface lipids as applied to the analysis of silks and seedling leaves of maize. PLOS
ONE, 12(7), e0180850. https://doi.org/10.1371/journal.pone.0180850
Ramirez-Rodrigues MM;Plaza ML;Azeredo A;Balaban MO;Marshall MR; (n.d.).
Physicochemical and phytochemical properties of cold and hot water extraction
from Hibiscus Sabdariffa. Journal of food science. Retrieved February 14, 2023, from
https://pubmed.ncbi.nlm.nih.gov/21535810/
R, K. (2017, October 17). Why do herbal tinctures contain alcohol? PPC Herbs. Retrieved
February 14, 2023, from
https://www.ppcherbs.com.au/herbal-tinctures-contain-alcohol/#:~:text=Alcohol%2
0is%20an%20excellent%20solvent,effectively%20extract%20the%20ingredients%20req
uired.
Hossain, M. M., Mondal, M., Morad, R. U., Uddin, N., Das, A., Hossain, M. S., Kamal, M. M.,
Islam, M. F., Wahed, T. B., &amp; Chowdhury, M. M. H. (2018, December 3).
Evaluation of bioactivities of methanol and petroleum ether extracts of Cassia
renigera seed - clinical phytoscience. SpringerOpen. Retrieved February 14, 2023,
from
https://clinphytoscience.springeropen.com/articles/10.1186/s40816-018-0091-x#:~:te
xt=Both%20methanol%20and%20petroleum%20ether,found%20in%20the%20methan
olic%20extract.
Thomas, J. (2020, October 15). Effect of Different Solvents on the Extraction of
Phytochemicals in Colored Potatoes.
https://www.scirp.org/journal/paperinformation.aspx?paperid=103615

(Pdf) Extraction of Natural Dyes from Plants - Researchgate.

https://www.researchgate.net/publication/329058662_Extraction_of_Natural_Dyes_from_Pl
ants.

Stauffer, D. Aharony A. (2003). Percolation. ScienceDirect. Retrieved February 14, 2023, from
‌https://www.sciencedirect.com/topics/chemistry/percolation#:~:text=Percolation%20is%20an%2
0exhaustive%20extraction,solvent%2C%20thus%20reducing%20solvent%20consumption.
 Experiment No. 5
PRELIMINARY SCREENING OF PLANT CONSTITUENT

SCHEMATIC DIAGRAM: (sue) and (roxeen)

1. What is the purpose of preliminary screening of plant constituents? (marianne)

The preliminary screening is used to identify bioactive compounds that are unknown in
plant extract; thus, it provides an overview of the class of compounds contained in the
plant being analyzed. The information gained from preliminary screening could be
significant to the synthesis of complex active components of chemical compounds
found in the plant. This can also be used to identify active compounds that cause toxins
apart from active compounds that are beneficial to human health. Hence, this causes
preliminary screening of plant constituents before further analysis.

2. Why is 80% ethanol used in preparing plant extracts? jenny

There is a lot of strong alcohol that can be used in preparing plant extracts, but ethanol
alcohol is most likely to be used. According to Sultan et al.., 2009 , 80 % ethanol that has
a high concentration are highly effective and safe for plant extract. Ethanol is added to
the plant for it to extract the phenolic derivative antioxidant compounds from natural
ingredients with good results by retrieving a large amount of oil from the plant.

3. Give the significance of each active plant constituents in pharmacy and medicines.
(jicelle) PACHECK OR PAADD NALNG PO
● Alkaloids-used as cardiac depressants, antihypertensive, anti-leukemic,
analgesic, nerve stimulants and local anesthetic
 ● Steroids-used as anti cholesterol, antibacterial, antineoplastic and growth
hormone regualator
● Anthraquinone Glycosides-used as laxative, antimalarial and antineoplastic
● Flavonoids-have anti-inflammatory and anti-viral properties
● Saponin-used in producing synthetic hormones, and has anti cholesterol,
anti-inflammatory, anti-viral, and anti-parasitic properties
● Cyanogenic Glycosides-has anticarcinogenic activity, thereby, used in
antineoplastic drugs.
● Carbohydrates-produces mucilage and gums which are process to create
medicines that treats dry skin and tissues
● Proteins-has antimicrobial, antioxidant and neurologic activity
● Volatile oils-produces essential oils and some are strong antiseptic.

4. What are primary, secondary and tertiary alkaloids? (cindy)

Alkaloids are often classified on the basis of their chemical structure. Those alkaloids that
contain a ring system called indole are known as indole alkaloids. Primary alkaloids are
formed when one hydrogen atom in ammonia is substituted by an alkyl or aromatic
group. Some examples of primary alkyl amines include amino acids and methylamine,
while primary aromatic amines include aniline. They are an important structural motif in
active pharmaceutical ingredients (APIs) and intermediates thereof, as well as members
of ligand libraries for either biological or catalytic applications. Secondary alkaloids are
those amines that have two organic substituents, either alkyl or aryl ones or both. They
are bound to the nitrogen together with one hydrogen. A common example includes
dimethylamine. Likewise, diphenylamine is an example of an aromatic amine. Thirtiary
alkaloids are which the nitrogen atom is directly bonded to three carbons of any
hybridization which cannot be carbonyl group carbons. General tertiary amine structure.
C = any carbon group except carbonyl. Applications of it include quaternary derivatives,
amine oxides and betaines which are used in household, industrial, and institutional
cleaners and disinfectants, wood treatment, personal care, oil field, and water treatment
end-use markets.

5. What are cardiac glycosides? (tristan)

Cardiac glycosides are medications used to treat people with heart failure, such as atrial
fibrillation and atrial flutter. They are commonly derived from foxglove plants such as
Digitalis lanata and Digitalis purpurea. A commonly prescribed cardiac glycoside,
Digoxin. Their mechanism of action is involved with the inhibition of the Na+ K+ ATPase
enzyme, also known as the sodium-potassium pump. The risk of having an overdose
 being too severe, they are not used as first-line for treatment for these conditions, but if
other medications do not work, they can be used but at lower doses for safety.

References: (APA 7)

Cardiac Glycosides. (2012). PubMed; National Institute of Diabetes and Digestive and Kidney

Diseases. https://www.ncbi.nlm.nih.gov/books/NBK548418/

Cardiac glycoside overdose: MedlinePlus Medical Encyclopedia. (n.d.). Medlineplus.gov.

https://medlineplus.gov/ency/article/002581.htm

Medicinal botany - Active plant ingredients. (n.d.). US Forest Service.

https://www.fs.usda.gov/wildflowers/ethnobotany/medicinal/ingredients.shtml
Medicinal plants and its pharmacological values. (2021, October 20). IntechOpen - Open
Science Open Minds | IntechOpen. https://www.intechopen.com/chapters/79065
Patel, V., & Patel, R. (2016). Journal of Chemical and Pharmaceutical Research | JOCPR.
https://www.jocpr.com/articles/the-active-constituents-of-herbs-and-their-plant-chemistr
y-extraction-and-identification-methods.pdf

Sultana B. , Anwar, F ., Ashraf M. (2009 July 15). Effect of Extraction Solvent/Technique on the
Antioxidant Activity of Selected Medicinal Plant Extracts. NCBI NLM. Retrieved February
19, 2023 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6254218/

Fardiyah, Q., Suprapto, Kurniawan, F., Ersam, T., Slamet, A., & Suyanta. (2020). Preliminary
phytochemical screening characterization of several medicinal plants extract from East
Java Indonesia. IOP Conf. Series: Materials Science and Engineering, 833(2020).
http://dx.doi.org/10.1088/1757-899X/833/1/012008

Alkaloid - an overview | sciencedirect topics. (n.d.). Retrieved February 19, 2023, from
https://www.sciencedirect.com/topics/food-science/alkaloid

Synthesis of sterically hindered primary amines by ... - ACS publications. (n.d.). Retrieved February
19, 2023, from https://pubs.acs.org/doi/10.1021/jacs.9b10871

Tertiary amine. Tertiary Amine - an overview | ScienceDirect Topics. (n.d.). Retrieved February
19, 2023, from https://www.sciencedirect.com/topics/chemistry/tertiary-amine
 Experiment No. 7
CARBOHYDRATES

1. Compare acid and enzymatic hydrolysis of starch. (tristan)

Acid Hydrolysis is a chemical process, and it’s is the cleavage of chemical
bonds in molecules with the addition of water in an acidic medium, while Enzymatic
hydrolysis is a biochemical process and it’s is the cleavage of chemical bonds in
molecules with the addition of water with the presence of an enzyme. During acid
hydrolysis of starch, it changes the structural and functional properties of starch without
changing its granular morphology, starch in an acidic medium, the glycosidic bond's
oxygen is attacked by the hydroxonium ion, which then hydrolyzes the connection. On
the other hand, during enzymatic hydrolysis of starch via the enzyme amylase, it
hydrolyzes starch, breaking down starch into smaller or shorter molecules, resulting in
maltose, which is then split by maltase into two glucose molecules.

2. Write the different stages in the acid hydrolysis of starch, and state the color obtained
with iodine at each stage. (roxeen)

a. Liquefaction - In this stage, starch is transformed into its soluble form, which results
in a blue-to-black color with iodine

b. Saccharification - The soluble form of starch is turned into glucose. Glucose does
not react with iodine which results in a red-to-yellowish color (color of iodine).

3. What are dextrins? How are they prepared? (cindy)

Dextrin is a generic term applied to a variety of products obtained by heating a

starch in the presence of small amounts of moisture and an acid. Dextrins can be made
from any starch and are generally classified as white dextrins, yellow (or canary) dextrins,
and British gums. They are more water-soluble and produce less viscous solutions or
dispersions than its parent starch. It is produced by combinations of slight
depolymerization (hydrolysis) and transglycosylation (molecular rearrangement). When
starch is partially hydrolyzed by the action of acids or enzymes (amylases), it is degraded
to maltose, maltotriose, and an oligosaccharide called dextrin. One type of dextrin,
known as “limit dextrin,” is one of the products after digestion with amylase. Dextrin is
prepared by heating and drying starch and then it may be treated with starch with HCl
(hydrochloric acid) to produce a colorless to yellowish, tasteless, and odorless powder
which, when mixed with water, forms a strong adhesive paste. Since it is a dry process,
recovery of water-soluble materials is simpler than with aqueous fluidity and oxidized
 starches. Depending on reaction conditions, greater or lesser amounts of three reactions
will occur: (a) hydrolysis; (b) transglycosidation; and (c) repolymerization.

4. What is meant by an achromatic point? What is its significance? (gwane)

Achromatic points are the chromaticity of rays that, under a specific illuminant
situation, do not appear red, green, blue, or yellow. Surface color saturation and hue are
assessed concerning an achromatic point of the same lightness, whereas the
achromatic point under a colored illuminant depends on the lightness under the test
(Kuriki, 2018). The achromatic point is independent of the test stimulus's intensity,
indicating that the gain adjustment factors, or "von Kries coefficients," are invariant or
independent of the test color's relative luminance with the background. Using brightness
dependency of the achromatic point locus resulted in a 40% improvement in the
assessment of changes in color appearance (Kuriki, 2015).

5. Enumerate sources of sugars. Specify sugars present. (suue)

● Corn syrup, Honey, Fruits, Vegetables, and Honey- Glucose and Fructose
● Maple syrup and sugar cane- Sucrose
● Dairy products (cows milk)- Lactose
● Molasses, Wheat and Barley- Maltose
● Rice straws and Corncobs- Xylose
● Potato, Corn, Cassava- Starch

6. Compare sugar with starch in terms of classification, color, taste, and appearance.
jenny

Starches and sugars are the two different categories of carbohydrates. Starch is
categorized as a complex carbohydrate similar to polysaccharides, whereas sugars are simple
carbohydrates like mono- and oligosaccharides. Despite the fact that they are both white,
starch has no flavor, while sugar is sweet. In their physical condition, sugar is a crystalline solid,
whereas starch is an amorphous solid. Starches are somewhat soluble to insoluble in cold water
and soluble in hot water, whereas sugar is completely soluble in water.

7. Discuss the principle involved in each of the aforementioned chemical tests for
carbohydrates. Include equations when necessary. (jicelle)
● Molisch test
 The test is based on the dehydration of the carbohydrates or sugar in an acidic
environment(in the presence of sulfuric acid) to form furfural derivatives, then
undergoing condensation with alpha-naphthol forming hydroxymethylfurfural
derivatives (purple-colored furfural)
● Fehling’s test
This test is based on the presence of free aldehydes or ketones which reduces the
copper sulphate to cuprous oxide
● Barfoed’s test
This test is based on copper reduction, it is performed to distinguish
monosaccharides from disaccharides. The cupric ions from the Barfoed’s reagent
are converted to cuprous ions with the formation of copper oxide. Thus, a mixture
with a red precipitate is formed for reducing monosaccharides.
● Osazone test
This test is based on the condensation-oxidation reaction between the
phenylhydrazine of the osazone reagent and the free carbonyl of carbohydrates
forming an osazone, producing crystal forms.

● Seliwanoff’s test
This test is used to differentiate sugars that contain ketohexoses from aldohexoses.
The ketose undergoes dehydration in the presence of concentrated HCl to form
5-hydroxymethylfurfural. The 5-hydroxymethylfurfural then reacts with resorcinol to
produce a cherry red complex. Aldohexoses react more slowly and produce a
faint pink-colored complex instead of cherry red.
● Aniline acetate test
This test is specifically for pentose. The furfural (from the conversion of pentose in
the presence of HCl) reacts strongly with the aniline forming a bright pink color.
● Tollens test
 This test is also known as the silver mirror test based on its positive result which is
metallic silver. This test is given by the redox reaction between the silver ions of the
tollen's reagent and the reducing sugars.
● Mucic acid test
The test is used specifically for lactose and galactose. Galactose-containing
sugars interact with an acid substance such as HNO3, the acid will oxidize the
terminal groups of the sugar, forming insoluble and clear crystals of
meso-galactaric acid or galactaric acid (mucic acid).
● Iodine test
This test is based on the absorptive properties of large polysaccharides, on which
the iodine forms complex between the helical chains of the polysaccharide, this
produces the appearance of blue coloration in the solution.
Test for Sugar Reagent with Principle Positive Result

*Physical Test Color

Odor
Taste
Appearance
Solubility

Molisch Test Dehydration of pentoses and hexoses by sulfuric acid to form furfural / red -cum-violet ring
hydroxymethylfurfural
furfural / hydroxymethylfurfural + a-naphthol = purple or violet

Fehling’s Test Fehling’s A: cupric sulfate pentahydrate Yellow to brick red

Fehling’s B: Alkaline sodium potassium tartrate (Rochelle salt) + NaOH
precipitate
sugar reduces cupric ion along with a resultant oxidation of the
carbonyl group of the sugar, to yellow or red cuprous oxide and is
precipitated.

Barfoed’s Test complex mixture of sodium carbonate, sodium citrate and copper(II) sulfate Red precipitate
Pentahydrate (similar to Benedict’s but lower pH about 4.5 and heating time
reduce to 2-3 minutes

Osazone have a characteristic shape under the light microscope Crystals under the
Phenyl hydrazine reacts with carbonyl group (C=O) of the sugar to form phenyl
microscope
hydrazone, which then
reacts with two further molecules of phenyl hydrazine to form the osazone.

Seliwanoff’s AKA Resorcinol HCl Reaction Red color

Ketose + concentrated HCl = 5-hydroxymethylfurfural
Test
5-hydroxymethylfurfural + resorcinol = red product with 2 minutes
*aldohexoses reacts more slowly

Aniline 1. Dip a strip of filter paper in aniline acetate (Aniline + glacial acetic acid) Red color
solution.
Acetate Test
2. Dry it and expose to fumes from sample with HCl which is heated to
produced and record the time for
the development of pink color.
▪ Hexose + Hot HCl → hydroxymethyl furfural (less volatile)
▪ Pentose + Hot HCl → Furfural (more volatile) + aniline → bright red/pink color

Tollen’s Test ammoniacal silver nitrate solution Black mirror

Tollens reagent is a colorless, basic, and aqueous solution containing silver ions
coordinated to ammonia,
forming a diaminesilver(I) complex [Ag(NH3)2]+
The aldehyde is converted into carbon dioxide while the Ag+ ions are reduced
to Ag metal, which leaves
a shiny appearance

Mucic Acid which concentrated HNO3 is heated along with an aldose sugar to give a Crystals under the
dicarboxylic
Test microscope
Acid
Nitric acid + terminal group of aldoses = carboxylic acid ====== mucic acid

Iodine Test a coiled polymer of α-amylose and amylopectin. Iodine interacts Deep blue color
with these coiled molecules and becomes bluish black
Glycogen, the common polysaccharide in animals, has a slight difference in
structure and produces
only an intermediate (pale brown) color reaction.
References: (APA 7)
Al-Sharifi, N. (n.d.). AL-RASHEED PHARMACY Dpt. 3rd year 1st semester Biochemistry Lab 2

Seliwanoff’s Test and Bial’s Test.

https://www.alrasheedcol.edu.iq/modules/lect/lect/1445-Seliwanoff%E2%80%99s

%20Test%20and%20Bial%20s%20test.pdf#:~:text=The%20formation%20of%20the%2

0cherry

Benedict’s test. (2021, March 2). BYJUS.

https://byjus.com/chemistry/benedicts-test/

Britannica. (2023, January 20). Starch | Definition, Formula, Uses, & Facts. Encyclopedia

Britannica. https://www.britannica.com/science/starch

Gibson, R. (2022, September 5). Tests for carbohydrates | Types, principles, apparatus. A

Level Biology. https://alevelbiology.co.uk/notes/tests-for-carbohydrates/#11-principle

Kuriki, I. (2018). A Novel Method of Color Appearance Simulation Using Achromatic Point

Locus With Lightness Dependence - Ichiro Kuriki, 2018. I-Perception.

https://journals.sagepub.com/doi/full/10.1177/2041669518761731#:~:text=The%20ac

hromatic%20point%20was%20considered,background%2C%20at%20the%20same%2

0time.

Kuriki, I. (2015). Lightness dependence of achromatic loci in color-appearance

coordinates. Frontiers in Psychology, 6. https://doi.org/10.3389/fpsyg.2015.00067

J.N. BeMiller (2003), in Encyclopedia of Food Sciences and Nutrition (Second Edition),

Types of Sugar. (2019a, May 28).

https://www.sugar.org/sugar/types/#:~:text=Sugars%20are%20classified%20in%20a,for%20

different%20foods%20and%20beverages.
 Wang, S., & Copeland, L. (2015). Effect of Acid Hydrolysis on Starch Structure and

Functionality: A Review. Critical Reviews in Food Science and Nutrition, 55(8),

1081–1097. https://doi.org/10.1080/10408398.2012.684551

‌
 Experiment No. 6
PRODUCTION OF WINE

Magsusulat ng qfr and SCHEMATIC DIAGRAM: (jicelle)

Questions:

1. Describe the principle behind wine production. (gwane)

The following are principles used in winemaking presented by Swift (2022):

● Harvest- Harvesting can start when the grapes are ideally phenolic and sugar ripe. When
grapes are harvested too early, unripe or "green," notes can be introduced into the wine,
while when grapes are harvested too late, the wine can become flabby or jammy. The
weather must be considered; for instance, if a storm is brewing, bringing in the grapes a
little underripe could be wiser rather than risking a chance.
● Crushing and pressing- Destemming the grapes as they arrive in the cellar will prevent
adding more tannin to the wine by removing all the grapes from the stems and using
them to create drainage channels that improve the effectiveness of the passing
procedure. The juice collected by pressing the grapes is combined with the free-run juice
to produce white wines. Besides the juice, also pressing releases flavor, fragrance, and
polyphenols from the cells on the inner surface of the grape skins. A traditional wine press
has a basket with slats spaced apart and a top screwed down to press the grapes.
Winemakers must establish a balance between quantity and quality when pressing since
it causes the release of bitter polyphenols.
● Fermentation- Wine is produced when yeast cells convert sugar into ethanol, often
known as ethyl alcohol. Numerous genetically diverse strains of yeast belong to various
species. Saccharomyces cerevisiae, which is effective at fermenting carbohydrates into
ethanol and resilient to high concentrations of ethanol waste product, is the yeast most
frequently employed in the production of wine.
● Maturation- After fermentation, the simplest wines are almost immediately bottled.
Nonetheless, a post-fermentation maturation time is beneficial for the majority of wines.
After fermentation, chardonnay can be kept in contact with lees, mostly made up of
extinct yeast cells. Lees once again enhances the wine's richness and depth while
introducing yeast, bread, brioche, and biscuit elements. Oak barrels are frequently used
to age wine because they are waterproof but permit little oxygen to permeate the wine.
Wine can be harmed by too much oxygen. However, a small amount also has many
positive effects, such as improving color stability and intensity, eliminating vegetal and
reductive flavors, and softening tannins.
2. Give 10 products of fermentation and the organism producing each. (tristan)

3. Outline the Emben-Meyerhoff scheme for the formation of ethanol in yeasts. (marianne)

4. Why is the strength of alcoholic solutions directed by the USP/NF to be determined at

15.56°C? (roxeen)
Alcohol content should not be less than 92.3 percent and not more than 93.8 percent in
terms of weight. In terms of volume, alcohol content should not be less than 94.9 percent
and should not be more than 96 percent at 15.56 degrees celsius.

5. What is meant by proof and gallon proof when referring to alcohol or alcoholic
beverages? (gwane)

The proof is twice the volume of alcohol (ethanol). For instance, a whiskey with a
proof of 100 has 50% alcohol. 80-proof signifies 40% of the liquid is alcohol, while anything
120-proof is 60% alcohol (Valley, 2012).
The amount of ethyl alcohol in a given volume of liquid is measured as a "proof
gallon." A proof gallon has the same volume as a wine gallon but only contains 50%
ethanol. In practice, the remaining 50% can be any liquid as long as it is not ethanol.
Nonetheless, it is usually water (Galbiati, 2019).

6. If an alcohol beverage is 70% strong, what is the proof strength? Show computation.
(marianne)
7. Enumerate the types of wine and their respective sources. (suue)

8. Explain the physiologic action of alcohol both in small and large amounts. (cindy)
Alcohol is a substance that is consumed by individuals for leisure. It has several adverse
effects when consumed in high amounts. These include impairments in motor
coordination, memory formation, learning, and severe problems like chronic health
conditions. When the body consumes alcohol, several changes begin to take place
beginning with the very first sip. In fact, the first part of the body affected by alcohol is
the brain due to alcohol affecting neurotransmitters. Alcohol interferes with the brain's
communication pathways, and can affect the way the brain looks and works. These
disruptions can change mood and behavior, and make it harder to think clearly and
move with coordination.Drinking a lot over a long time or too much on a single occasion
can damage the heart, causing problems including high blood pressure and stroke. It
causes the pancreas to produce toxic substances that can eventually lead to
pancreatitis, a dangerous inflammation and swelling of the blood vessels in the pancreas
that prevents proper digestion. Drinking too much can weaken your immune system,
making your body a much easier target for disease. Chronic drinkers are more liable to
contract diseases like pneumonia and tuberculosis than people who do not drink too
much. Drinking a lot on a single occasion slows your body’s ability to ward off infections
– even up to 24 hours after getting drunk.

9. Why is alcohol not used in anesthetics anymore? jenny

Alcohol has been known for having a properties of anesthetic and analgesic for several
thousand years but upon introducing its use into anesthetic practice it does not become
successful because its main use was as a stimulant for resuscitation. Hence , alcohol is not
use anymore as anesthetics.

10. Discuss the significance of fermentation in pharmaceutical processes. Cite examples.

(roxeen)
Fermentation is important in the pharmaceutical industry because it helps in creating a
wide range of drugs for illnesses and disease. Microbial Fermentation is the type of
fermentation used in the synthesis of drugs such as antibiotics and anticancer cytotoxic
drugs. Most antibiotics are created through fermentation since microorganisms produce
antibiotic materials during fermentation which are then isolated for the production of
antibiotics. An example would be natural penicillins which are isolated from the
fermentation of Penicillium chrysogenum.
References: (APA 7)

Chapman, R. A., & Plaat, F. (2009, February 1). Alcohol and anaesthesia. Continuing
Education in Anaesthesia, Critical Care & Pain; Oxford University Press.
https://doi.org/10.1093/bjaceaccp/mkn045
Galbiati, S. (2019, December 6). What is the difference between a “wine gallon” and a
“proof gallon”? Culinary Solvent; Culinary Solvent.
https://culinarysolvent.com/blogs/ethanol/what-is-the-difference-between-a-wine-
gallon-and-a-proof-gallon

Principles of winemaking. (2022, June 5). Atlas Swift.

https://www.atlasswift.co.za/blogs/articles/principles-of-winemaking

Valley, N. (2012, May 21). What Does Proof Mean In Alcoholic Beverages? Duffy’s Napa
Valley Rehab; Duffy’s Napa Valley Rehab.
https://www.duffysrehab.com/about/blog/what-does-proof-mean-in-alcoholic-bev
erages/
Abdallah, Qusai & Nixon, B. & Fortwendel, Jarrod. (2016). The Enzymatic Conversion of
Major Algal and Cyanobacterial Carbohydrates to Bioethanol. Frontiers in Energy
Research, 4. https://doi.org/10.3389/fenrg.2016.00036
U.S. Department of Health and Human Services. (n.d.). Alcohol's effects on the body.
National Institute on Alcohol Abuse and Alcoholism. Retrieved March 12, 2023, from
https://www.niaaa.nih.gov/alcohols-effects-health/alcohols-effects-body#:~:text=Alcohol
%20interferes%20with%20the%20brain's,clearly%20and%20move%20with%20coordination.

Bistami, N. A., &amp; Chiprasov , A. (n.d.). Take online courses. earn college credit.
Research Schools, Degrees &amp; Careers. Study.com | Take Online Courses. Earn
College Credit. Research Schools, Degrees &amp; Careers. Retrieved March 12, 2023,
from
https://study.com/academy/lesson/the-effects-of-alcohol-physiological-behavioral-long-t
erm.html
Chapman, R. A., & Plaat, F. (2009, February 1). Alcohol and anaesthesia. Continuing
Education in Anaesthesia, Critical Care & Pain; Oxford University Press.
https://doi.org/10.1093/bjaceaccp/mkn045
 Experiment no. 8
EXTRACTION OF CITRIC ACID FROM LEMON

SCHEMATIC DIAGRAM- jenny&mariane

QFR
1. Differentiate the properties of citric acid and tartaric acid. TABULATE
Tristan

Properties (Physical and Citric Acid (C6H8O7) Tartaric Acid (C4H6O6)

Chemical)

State Solid Solid

Color White Crystal White Powder

Odor Characteristic Odorless Characteristic Odor

Flammability Non-luminous, non-smoky Non-luminous, non-smoky

flame flame

Solubility Completely miscible in Completely miscible in

water water

Taste Less sour Sour

Molecular weight 192.123 g/mol 150.086 g/mol

General:

Weak organic acid, Organic acid which

naturally occurs in citrus naturally occurs in grapes,
fruits such as limes, lemons, found in grapes and
and pineapples grape-based wine.

Also occurs in apricots,

oranges, bananas,
avocados, and tamarinds

Antioxidant, Adds sour Antioxidant, Adds sour

taste to food and taste to food and
carbonated drinks) carbonated drinks

Triprotic (3 ionizable/acidic Diprotic (2 ionizable/acidic

hydrogen atoms) hydrogen atoms)
 Natural Additive in food Natural Additive in food

Preservative(inhibit Preservative(inhibit less

bacterial growth) bacterial growth)

E number E330 E334

2. Provide natural sources of oth citric and tartaric acid

Jicelle
NATURAL SOURCES OF CITRIC ACID
● Lemons - Citrus limon
● Limes - Citrus aurantifolia
● Oranges - Citrus sinensis
● Tomato - Solanum lycopersicum
● Grapefruits -Citrus paradisi

NATURAL SOURCES OF TARTARIC ACID

● Strawberry - Fragaria ananassa
● Banana - Musa balbisiania
● Tamarind - Tamarindus indica
● Mango - Mangifera indica
● Cherry - Prunus avium

3.
Suue (citric), cindy(tartaric)

Citric acid
Citric acid is important in the field of pharmacy. Citric acid is a commodity chemical
processed and widely used around the world as an excellent pharmaceutical excipient.
It offers many significant properties such as biodegradability, biocompatibility,
hydrophilicity, and safety. It offers many uses such as flavoring agent, sequestering
agent, and buffering agent . It is used as a cosmetic ph adjustment and a metal ion
chelator. It is also an ingredient to improve the taste of syrups, elixirs and solution. Citric
acid is also utilized as a release modifying agent, capping, and coating agent.
Furthermore, it can be employed as an acidulant for mild astringent.
 Due to many important applications of citric acid in pharmacy, citric acid is a highly
valuable asset in pharmaceutical drug making.

Tartaric acid was first isolated from potassium tartrate. Tartaric acid and derivatives are
used widely in pharmaceutical manufacturing. One example is that tartaric acid has
been used to make effervescent salts, and when combined with citrate can improve the
taste of oral medications.It is used in various formulations for antibiotics, effervescent
powders and tablets, drugs for heart diseases, therapeutic compounds and excipient in
the preparation of some medicines.
It is used to improve the taste of oral medications
It is used to chelate metal ions such as magnesium and calcium
It is used in recipes as a leavening agent along with baking soda
It is used as an antioxidant
It is as one of the important acids in wine
It is used in foods to give a sour taste
It is sometimes used to induce vomiting
It is used to make silver mirrors
In its ester form, it is used in the dyeing of textiles
It is used in the tanning of leather
It is used in candies
In its cream form, it is used as a stabilizer in food.
4. Explain the significance of citric and tartaric acid in medicine.
Roxeen(citric), gwane(tartaric)

Citric acid is an acid found in an acid found in citrus fruits which is used widely in the
medical field. It is an anti-chelation additive in blood collection tubes and is used in the
formulation of different pharmaceutical products. It is also a key intermediate to metabolism.
Some creams contain citric acid which helps in clearing up skin infections. Orally administered
drugs containing citric acid decrease the acidity of urine. It helps prevent formation of kidney
stones and metabolic acidosis.

Chemically, tartaric acid plays a significant role by bringing the pH of fermenting "must"
down to a level below which many spoilage bacteria cannot survive and acting as a
preservative after fermentation.
Moreover, Tartaric acid is used as a primary raw material to create emulsifiers, which are
a vital component of improved bread, which is its most common use. A significant tartaric acid
salt called potassium hydrogen tartrate, also known as "cream of tartar," is used as an acidulant
in baking powder and sugar confections.
REFERENCES:
Admin. (2022, August 3). Tartaric acid (C4H6O6) - structure, Molecular Mass, properties &amp;
uses. BYJUS. Retrieved March 20, 2023, from https://byjus.com/chemistry/tartaric-acid/

Booth, S. (2019, September 11). What is citric acid? WebMD.

https://www.webmd.com/diet/what-is-citric-acid
Citric acid. (n.d.). DrugBank Online | Database for Drug and Drug Target Info.

https://go.drugbank.com/drugs/DB04272

Lakna. (2018, May 4). Difference Between Tartaric Acid and Citric Acid. Pediaa.com.

https://pediaa.com/difference-between-tartaric-acid-and-citric-acid/

Lambros, M., Nicolaou, M., Qinqin, F., & Tran, T. (2022, April 30). Citric acid: A Multifunctional

pharmaceutical excipient. PubMed Central (PMC).

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9148065/#:~:text=Citric%20acid%20is%2

0used%20to,correlated%20with%20bitterness%20%5B54%5D

Monoprotic and Polyprotic acids. (2023). Study.com.

https://study.com/academy/lesson/monoprotic-and-polyprotic-acids.html#:~:text=Polyp

rotic%20acids%20can%20either%20be

Nangare, S., Vispute, Y., Tade, R., Dugam, S., & Patil, P. (2021). Pharmaceutical applications of
citric acid. Future Journal of Pharmaceutical Sciences, 7(1).
https://doi.org/10.1186/s43094-021-00203-9
Stevens, H. (1924). A Simple Distinction between Citric and Tartaric Acids. Industrial & Engineering

Chemistry, 16(2), 155–155. https://doi.org/10.1021/ie50170a021

Tartaric acid. (2016, June 14). The Chemical Company.

https://thechemco.com/chemical/tartaric-acid/
Tartaric Acid - an overview | ScienceDirect Topics. (2008). Www.sciencedirect.com.

https://www.sciencedirect.com/topics/agricultural-and-biological-sciences/tartaric-acid

Ukaji, Y., & Soeta, T. (2012). 3.6 Acetogenin (Polypriopionate) Derived Auxiliaries: Tartaric

Acid. Comprehensive Chirality, 176–201.

https://doi.org/10.1016/b978-0-08-095167-6.00306-2