Acquisition of MS/MS spectra

x10 2 + TIC MS(all) Meta27_10_11_Comp1_110-r001.d

3

0
2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48
Counts (%) vs. Acquisition Time (min)

Extraction of TIC for MS

Level
x10 2 + TIC Scan Meta27_10_11_Comp1_110-r001.d
3

0
2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48
Counts (%) vs. Acquisition Time (min)

Extraction of MS profiles

Extraction of
x10 4 +ESI Scan (0.82-47.95 min, 1413 Scans) Frag=195.0V Meta27_10_11_Comp1_110-r001.d
MS/MS spectra
453.3493

475.3315

496.3471

3
121.0524

of differentially
235.1835
160.0625

315.0847

436.4384
293.1018

2 modulated
520.3472

592.5663
339.3483

468.4648

618.5823

679.5193
573.5241
395.4113
274.2784

415.2175

544.3456

compounds
1

0
100 125 150 175 200 225 250 275 300 325 350 375 400 425 450 475 500 525 550 575 600 625 650 675
Counts vs. Mass-to-Charge (m/z)

Statistical Analysis


Pi
tt
a

K
a
K P V p
Pi
a a h
p rt t a
h at t 
CC ka +ESI Product
a 356.0832: a
oo x10 3 20.58 V (356.0820[z=1] -> **) Meta27_10_11_Comp1_110-r001.d
Ion (20.58 min) Frag=195.0V CID@28.0
ri
at
356.0832

l l 1.25
[[ [[ [[ ti
P P KK VV
oo 1 ta
rr e
313.0236

]] ]] ]]
175.0776

rr 0.75
s
153.1290

198.9420

342.0683
115.0984

215.0774

aa 0.5
nn 0.25
gg
ee 0
100 125 150 175 200 225 250 275 300 325 350 375 400
ff Counts vs. Mass-to-Charge (m/z)
oo
r
Differentiallyrmodulated MS/MS Spectra
ff
compounds o o
l l
dd
cc
Fig. S1. Flowchart showing methodology h hfor statistical analysis of data and identification of compounds using MS/MS fingerprints.
aa
nn
gg
ee
ss
 x10 7 +ESI TIC Scan Frag=175.0V SERUM54-r001.d A: Pitta
2.1 1 1
2
1.9
1.8
1.7
1.6
1.5
1.4
1.3
1.2
1.1
1
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29
Counts vs. Acquisition Time (min)

x10 7 +ESI TIC Scan Frag=175.0V SERUM7-r001.d B:Vatta

1 1
1.6
1.5
1.4
1.3
1.2
1.1
1
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29
Counts vs. Acquisition Time (min)

x10 7 +ESI TIC Scan Frag=175.0V SERUM7-r002.d C: Kapha

1.4 1 1

1.3

1.2

1.1

0.9

0.8

0.7

0.6

0.5

0.4

0.3

0.2

0.1

0
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29
Counts vs. Acquisition Time (min)

Fig. S2. Extracted Total Ion Chromatogram (TIC) of (A) Pitta prakriti (B) Pitta prakriti and (C) Pitta prakriti in overlaid mode showing the
consistency and variations in data.
 DG(18:3(9Z,12Z,15Z)/20:2(11Z,14Z)/0:0)
21-Hydroxydydrogesterone glucuronide
10,11-Dihydro-20-trihydroxy-leukotriene B4
A: N-Docosanoyl taurine 20.11 min

Chenodeoxycholic acid glycine

B: L-Octanoylcarnitine 23.78 min

Hexanoylcarnitine
5-Methylcytosine
C: Nonanoylcarnitine 19.45 min

Sphinganine

23.38 min
x10 6 +ESI BPC Scan Frag=175.0V Met19_7_1149-r001.d

conjugate
1.55 1 C A 1

1.5

1(3)-glyceryl-6-keto-PGF1α
1.45

Pimelylcarnitine
1.4

1.35

1.3
1.25

1.2

1.15
1.1
1.05

PS(19:0/0:0)
1
0.95

0.9
0.85
0.8

0.75
0.7 B
0.65

0.6
0.55
0.5
Melatonin

0.45
0.4

0.35

0.3
Fig. S3. The extracted +ESI Base peak chromatogram (BPC) representing kapha (K), pitta (P) and vatta (V). The peaks of identified compounds are
0.25

being labeled.
0.2

Abbreviations:
0.15
0.1
A: N-Docosanoyl taurine
0.05
B: L-Octanoylcarnitine
0
C:0 Nonanoylcarnitine
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
Counts vs. Acquisition Time (min)
21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37
 A

Fig. S4. Pie charts of Go Biological process in prakrities analysed using metscape and Cluego modules of cytoscape 3.3 (A) Kapha (B) Pitta (C)
Vatta.
 Table S1: Proforma for collection of blood and prakriti assessment

National Research Institute of Basic Ayurvedic Sciences (NRIBAS), Kothrud, Pune- 411038

1. Name of Volunteer :……………………………………………………

2. Age/ Sex :……………………………………………………

3. Education :……………………………………………………

4. Occupation :……………………………………………………

5. Marital Status :………………………………………………..

6. Address :……………………………………………………

7. Religion :……………………………………………………

8. History
State of Health :……………………………………………………

Family History :……………………………………………………

Personal History :……………………………………………………

9. Body Factor: Performa for assessment of Prakruti (Psychosomatic constitution)

Pulse Rate /min: Blood pressure (mmhg): Systolic: Diastolic:

Physique:

1a Body frame Lean & Tall

Medium
Large
1b Musculature Thin
Fleshy
Bulky

2 Body Weight: Low / Below average

Medium / Normal
Overweigh

3 Skin type:

3a General Feel Dry

Premature wrinkles
Delicate / Normal
Oily / Glistening
3b Feel on touch Cold feel

Warm feel

3c Complexion Dark / Brownish

Fair complexion / yellowish red
Pale / whitish complexion
3d Sensitivity / Intolerance Allergic to dry weather

Intolerant to hot weather

Intolerant to humidity

4 Skin texture: Often / seasonally cracked

Thin & smooth
Thick & soft
5 Sweating: Scanty

Profuse

Moderate & Oily

6 Hair:
6a Feel Dry
Soft
Oily
6b Appearance of strands Straight stands

Wavy

Curled

6c Color Blackish brown.

Reddish / Graying trends
Black & Lustrous
6d Intensity Scanty / Thinly Distributed

Thick

Alopecla / baldness

7 Eyes:

7a Eye Lashes Scanty eye lashes

Moderate eye lashes
Thick / Fused eye lashes
7b Eye balls Small eye balls

Medium sized eye balls

Large sized eye balls

7c Color of sclera (white ball) Grayish sclera

Reddish sclera
Pale white sclera

7d Color of iris (Blackball) Brown / black Iris

Dull & Lusterless

Grayish yellow Iris

Bright

Blue Iris

Moist

7e Blinking Excessive
Moderate blinking
More or less stable

7f Vision Curious vision

Sharp & penetrating vision

Dreamy vision

8 Teeth & Gums:

8a Row Irregular and spaced

Regular & close
Regular & spaced

8b Gums Thin

Soft & Fleshy

Thick and healthy

8c Size of teeth Large / protruded

Moderate size
Small sized

8d Color of teeth Brown tinged

Having yellow tinge

White / shining

9 Lips:
9a Texture Dry/ cracked often
Soft / moist
Oil / glistening

9b Color of lips Bluish / brownish red

Reddish, with yellow tinge

Pale

10 Cheeks:

10 a Structure Sunken
Flattened
Rounded / Plump

10 b Feel on touch Dry / Rough

Smooth

Soft

11 Neck: Long
Medium
Short / stout
12 Chest: Flat / sunken

Moderate / well defined

Expanded / Rounded

13 Abdomen: Thin / sunken

Moderate / flat
Large / Bulging

14 Hips & Thighs: Thin & Slender

Moderate / fleshy

Heavy / Bulging

15 Musculature: Thin & Hard

Thick & well defined
Stout & ill-defined
16 Joints:

16 a Visibility Protruding / prominent

Just visible
Invisible
Fleshy

16 b Mobility Cracking sounds

Free mobility

Slightly restricted mobility

17 Nails:

17 a Thickness Thin
Moderately thick
Strong & thick

17 b Texture Rough / Brittle

Soft / Rubbery

Hard

17 c Color Bluish / grayish colored

Pink colored
 Pale colored

17 d Formation / symmetry Marked ridges

Well formed

Symmetrical

18 Appetite:

18 a Frequency Irregular / unpredictable

High frequency
Low frequency
18 b Quantity of intake Satiation with little food

Fairly good intake

Low intake

18 c Tolerance to fasting Unpredictable

Poor, can not miss a meal
Good tolerance
19 Digestion: Irregular / undefined

Good

Low but regular

19 b Tendencies of disturbance Gaseous belching

Acidic belching
Undefined discomfort
20 Liking to tastes: Sweet / Sour / Salty

Sweet / Bitter Astringent

Pungent / Bitter /Astringent

21 Evacuation / Bowel movement:

21 a Regularity Irregular frequency

Regular / punctual
Regular / untimely
21 b Consistency Hard stool / low water content

Soft stool / higher water content

Soft-moderate water content

22 Thirst:

22 a Frequency High & non persistent

High & persistent
Scanty
22 b Intensity Low & satisfied with less water

Need more volumes

Does not apply

23 Sleep:

23 a Duration Short (< 4 hours)

Moderate (4-6 hours)
Long (> 6 hours)
23 b Depth Superficial

Disturbed

Deep & Sound

24 Physical strength &

Endurance:

24 a Activity Level Very active

Moderately active
Poor active
24 b Physical stamina Low stamina

Medium stamina

Excellent stamina

24 c Endurance Early tiredness / low endurance

Moderate endurance
Good endurance
24 d Reflexes after exercise Feel thirsty after exercise

Feel hungry after exercise

No hunger / thirst after exercise

27 Movements / Gait: Fast & unsteady

Fast & steady
Slow & steady
28 Weather tolerance: Intolerance to cold & dry

Intolerance to hot & humid

Intolerance to cold & Humid

29 Voice:

29 a Audibility Hoarse
Sweet
Deep
29 b Pace & Intensity Rapid

Sharp / commanding

Prolonged

29 c Clarity
Unclear / penetrating
Clear / penetrating
Slurred / feeble
Speech / Talking
attitude:
30 a Tendency Talkative. Often a loose talk
Talkative. Always firm talk
Non-talkative / Reserved

30 b Consistency Drifting between topics

Impressive speech
Concentrating

30 c Pace & Intensity Fast & diffused

Sharp & aggressive

Soft, slow & steady speech

30 d Impressibility Poor & inconsistent

Good & consistent
Deep & Pleasant speech

31 Liking foods: Hot / Oily

Cold / wet

Hot / dry

33 Decision making: Indecisive / unpredictable

Fast & Calculated
Slow & measured

34 Inquisitiveness: Highly inquisitive

Moderately inquisitive

Non-inquisitive / unconcerned

35 Enthusiasm & perusal: Very enthusiastic

Enthusiastic
 Less enthusiastic
Vivacious
Strong minded
Calm and cool

36 Grasping: Fast & superficial

Fast & medium in depth

Slow & go in to depth

37 Memory: Forget quickly / less

Short lived / Moderate
Deep & Long lived / good

38 Anger: Quick & Unstable

Quick & sustained

No anger

39 Anxiety / depression patterns: Prone to anxiety & depression

Prone to anxiety
Prone to depression

40 Response to problems:

40a Behavior Worried / Confused

Agitated
Calm

40b Solutions Often a wrong solution

Right & firm solution

Slow-but right solution

42 Tolerance to pain: Low -pain threshold

Medium pain threshold
Good pain threshold
46 Mind -set: Active-always
Restless
Disciplined
Aggressive
Calm
Stable

47 Hobbies: Creative / Educative

Art & Culture

Ornamental / fanciful

Adventure / outdoor games

Sedentary / philosophical

Indoor games

48 Lifestyle: Impulsive spending

Planned spending / flamboyant
Money saving / miser

49 Religious faith: Frequently changing

Fanatic

Permanent

50 Temperament in general: Impatient

Timid
Jealous
Thankless some time
High respect for other
Low self esteem
Speak cleverly
Act thoughtlessly
Adventurous at first but, retract
fast
Like to wander
 Intolerant
Greedy
Fearless
Poor respect to others
High / over self esteem
Consistently adventurous
Clever and cunning
Short tempered
Thankless, mostly
Brave-but, non-adventurous
Calm & quiet
Respect only to deserving
Patient
Forgiving
Thoughtful
Closefisted
Non-greedy / tolerant
Thankful, always

51 Tongue: Darkish
Coppery

Clean

52 Tendons: Visible
Not visible

53 Veins: Prominent and reticulated

Deep

54 Body Odour: Strong

Mild
Very less

55 Body weight changes: Gain and lose easily

Difficulty in gaining
Gain easily and lose with difficulty
Stable

56 Weather: Prefer Have Health Problem

Cold
Warm
Dry
Moist
Moderate
Stable

57 Season: Prefer Have Health Problem

Summer
Early winter
Late winter
Autumn
Spring
Rainy season
None

58 Speed: Quickly Moderately Slowly Variably

Memorizing
Forgetfulness
Recalling
Initiation
Making new friends
Anger
Irritability

Declaration by volunteer:

I do hereby give consent that I am donating my blood sample for research purpose with my own desire.

Sign of volunteer
Table S2: The characteristics used for discrimination among prakrities
1) Vatta prakriti - thin and narrow body frame, weakly developed body build, with irregular appetite, food and bowel habits, difficulty in
gaining weight, quick at physical activities, dry skin and hair, and less tolerance for cold temperature
2) Pitta prakriti - moderately developed build, high frequency of appetite and thirst, good digestive power, perspiration tendency higher than
normal, tolerance for cold weather, moderately mobile with moderate physical strength
3) Kapha prakriti - broad body frames with well developed body build, tendency to gain weight, low appetite and digestion, preferred to be less
mobile, less forgetful and with good healing power and cool temperament